國立臺南大學教師基本資料

基本資料
姓名 鄧燕妮
系所 生物科技學系
職稱 教授
校內分機 795
傳真
辦公室/研究室 格致樓C114, C102-2
E-mail tengyenni1968@gmail.com
網址
專長/研究領域 生物科技, 分子生物, 人類遺傳, 生物檢測
 

畢業學校國別主修學門學位修業期間
國立陽明醫學大學 中華民國 生物化學所 碩士 1991/09~1993/07
國立臺灣師範大學 中華民國 生物學系博士 1995/09~1999/12

服務機關部門 / 系所職稱服務期間
行政院衛生署預防醫學研究所 病毒組助理研究員 1993/09~1995/09
嘉南藥理學院嬰幼兒保育系講師1999/08~2000/01
嘉南藥理科技大學嬰幼兒保育系助理教授2000/02~2003/07
嘉南藥理科技大學嬰幼兒保育系副教授2003/08~2006/07
嘉南藥理科技大學生物科技系(所) 副教授 2004/08~2010/07
中央研究院 生物醫學研究所 國內學人短期訪問研究 2008/06~2008/09
國立臺南大學生物科技系副教授2010/08~2013/07
國立臺南大學生物科技系教授2013/08~迄今

著作
名稱Isochromosome of Yp in a man with Sertoli-cell-only syndrome.Lin YH, Chuang L, Lin YM, Lin YH, Teng YN, Kuo PL.Fertil Steril. 2005 Mar;83(3):764-6.
年度2005
類別期刊論文
摘要OBJECTIVE: To address phenotype/genotype correlation in a man with i(Y)(p10). DESIGN: Case report. SETTING: University-based reproductive genetics laboratory. PATIENT(S): A 27-year-old azoospermic man with i(Y)(p10), relatively normal stature, and testicular Sertoli-cell-only syndrome. INTERVENTION(S): Testicular biopsy, cytogenetic study, Y-chromosome deletion mapping analysis, fluorescence in situ hybridization (FISH). MAIN OUTCOME MEASURE(S): Expression of Y-chromosome genes. RESULT(S): We have identified one azoospermic man with i(Y)(p10) of 312 Taiwanese men presenting with a severe spermatogenic defect. Y-chromosome deletion mapping analysis confirmed deletions of all Yq sequences, including a putative growth controlling gene. Fluorescence in situ hybridization (FISH) analysis showed duplication of Yp material. The patient had normal stature considering midparental height. He also had no germ cells in the testicular tissue (Sertoli-cell-only syndrome) resulting from the loss of azoospermia factor in Yq. CONCLUSION(S): Among structural rearrangements of the Y-chromosome, the isochromosome of Yp occurs very rarely. This case is the first reported case of an isochromosome Yp with a detailed description of testicular histology and body height.
關鍵字
名稱Messenger RNA transcripts of the meiotic regulator BOULE in the testis of azoospermic men and their application in predicting the success of sperm retrieval.Lin YM, Kuo PL, Lin YH, Teng YN, Nan Lin JS.Hum Reprod. 2005 Mar;20(3):782-8. Epub 2004 Dec 9.
年度2005
類別期刊論文
摘要BACKGROUND: Testicular sperm retrieval can lead to paternity for azoospermic patients with spermatogenic failure. The human BOULE gene, a meiotic regulator of germ cells, is a gene whose altered expression may be associated with sterility. We determined the levels of BOULE transcripts in the testes of azoospermic patients, and evaluated the relationship between BOULE transcript levels and patients testicular phenotypes, clinical parameters and sperm retrieval results. METHODS and RESULTS: BOULE transcript levels in the testes of 41 azoospermic patients were examined by quantitative competitive-reverse transcription-polymerase chain reaction. A significant decrease in BOULE transcript levels was detected in patients with spermatogenic failure, and BOULE transcript levels progressively decreased with increasing severity of testicular failure. BOULE transcript levels did not correlate with the serum hormone parameters measured. Significantly higher BOULE transcript levels were detected in 19 patients with successful sperm retrieval than in 12 patients with failed sperm retrieval. When using a cut-off value of 0.5 for BOULE transcript ratio to predict the success of sperm retrieval, both the sensitivity and specificity value were 100%. CONCLUSIONS: We suggest the BOULE transcript plays an important role in human spermatogenesis and that the levels may predict the presence of testicular sperm in patients with spermatogenic failure.
關鍵字
名稱Identification of ten novel genes involved in human spermatogenesis by microarray analysis of testicular tissue.Lin YH, Lin YM, Teng YN, Hsieh TY, Lin YS, Kuo PL.Fertil Steril. 2006 Dec;86(6):1650-8. Epub 2006 Oct 30.
年度2006
類別期刊論文
摘要OBJECTIVE: To identify novel genes that are down-regulated in the testicular tissue of infertile men. DESIGN: Prospective study. SETTING: University-based reproductive clinics and genetics laboratory. PATIENTS: Nine patients with normal spermatogenesis, and 15 patients with maturation arrest (MA) or Sertoli cell-only syndrome (SCOS). INTERVENTION: Testicular samples of patients with the same histology were pooled for complementary DNA (cDNA) microarray analysis. MAIN OUTCOME MEASURE: Novel, down-regulated genes. RESULTS: In total, 300 genes were significantly down-regulated in SCOS or MA samples, and 10 novel sterility-related genes were identified. Of the 10 novel genes, 6 genes (Hs.126780, Hs.553658, Hs.274135, Hs.268122, Hs.531701, and Hs.171130) encode proteins with predictable functional domains, and all these functional domains are believed to correlate with spermatogenesis and/or spermiogenesis. Conversely, the other 4 genes (Hs.351582, Hs.407480, Hs.552781, and Hs.355570) do not encompass known functional domains. Two genes (Hs.407480 and Hs.552781) lack mouse orthologues. Most novel genes showed a testis-specific expression pattern in both mice and humans. Reverse transcription-polymerase chain reaction (RT-PCR) showed three distinct types of developmental stage-dependent expressions of message ribonucleic acid (mRNA) for these novel genes in murine testes. CONCLUSION: These 10 novel genes provide targets to elucidate novel pathways involved in human spermatogenesis.
關鍵字
名稱Association of DAZL haplotypes with spermatogenic failure in infertile men.Teng YN, Lin YM, Sun HF, Hsu PY, Chung CL, Kuo PL.Fertil Steril. 2006 Jul;86(1):129-35. Epub 2006 May 30.
年度2006
類別期刊論文
摘要OBJECTIVE: To identify novel DAZL single nucleotide polymorphisms (SNPs) and to compare allele/genotype frequencies, linkage disequilibrium (LD) characteristics, and DAZL haplotypes between fertile and infertile men. DESIGN: Prospective case study. SETTING: University genetics laboratory and reproductive clinics. PATIENT(S): Two hundred thirty-one infertile men and 191 men with proven fertility. INTERVENTION(S): Single strand conformation polymorphism and sequence analysis for DAZL gene polymorphism screening were done for all subjects enrolled. MAIN OUTCOME MEASURE(S): Novel SNPs, allele/genotype frequencies, LD characteristics, and DAZL haplotypes between fertile and infertile men. RESULT(S): Five SNPs were identified: 260A>G, 386A>G, 520+34c>a, 584+28c>t and 796+36g>a. SNP 386A>G was significantly associated with spermatogenic failure and was mainly heterozygous in infertile patients. The major haplotypes in infertile men were AACTA (45.8%), followed by AACCG, AAATA, AAACG, and GGACG for 260A>G/386A>G/520+34c>a/584+28c>t/796+36g>a. The major haplotypes for the control subjects were AACCG (41.7 %), AAATA, and AACTA. Of all haplotypes, five showed significant differences in frequency between infertile men and control subjects. Haplotypes AACTA, AAACG, and GGACG were overtransmitted in patients with spermatogenic failure, whereas haplotypes AACCG and AAATA were undertransmitted in these patients. CONCLUSION(S): Our study suggests the association of autosomal DAZL haplotypes with human spermatogenic failure.
關鍵字
名稱Association of spermatogenic failure with decreased CDC25A expression in infertile men.Cheng YS, Kuo PL, Teng YN, Kuo TY, Chung CL, Lin YH, Liao RW, Lin JS, Lin YM.Hum Reprod. 2006 Sep;21(9):2346-52. Epub 2006 May 23.
年度2006
類別期刊論文
摘要BACKGROUND: DAZ gene family is crucial for human spermatogenesis that requires the precise co-ordination of cell cycle events. CDC25A is recognized as the downstream substrate of DAZ gene family and is thought to function on the M-phase regulation of cell cycles. We investigated the expression profiles of CDC25A in the testes of infertile men and evaluated the relationship between CDC25A levels and testicular phenotype, clinical hormonal parameters and sperm retrieval results. METHODS: The protein and mRNA transcript levels of CDC25A in the testes of 40 azoospermic men were determined by immunohistochemistry and quantitative real-time-PCR. CDC25A in human spermatozoa was investigated by western blotting and immunofluorescence staining. RESULTS: The CDC25A protein was expressed mainly in spermatocyte, spermatid and spermatozoa. CDC25A transcript levels were significantly decreased (P 0.0009) in patients with spermatogenic failure, especially in men with meiotic arrest and Sertoli cell-only syndrome. Significantly higher CDC25A transcript levels were detected in patients with successful sperm retrieval than in patients with failed sperm retrieval (P 0.005). CONCLUSIONS: Decreased CDC25A is associated with spermatogenic failure and failed sperm retrieval in infertile men. Further studies are necessary to explore the functional roles of CDC25A in human spermatozoa.
關鍵字
名稱Uniform deletion junctions of complete azoospermia factor region c deletion in infertile men in Taiwan.Hsu CC, Kuo PL, Chuang L, Lin YH, Teng YN, Lin YM.Asian J Androl. 2006 Mar;8(2):205-11.
年度2006
類別期刊論文
摘要AIM: To determine the deletion junctions of infertile men in Taiwan with azoospermia factor region c (AZFc) deletions and to evaluate the genotype/phenotype correlation. METHODS: Genomic DNAs from 460 infertile men were examined. Bacterial artificial chromosome clones were used to verify the accuracy of polymerase chain reaction. Deletion junctions of the AZFc region were determined by analysis of sequence-tagged sites and gene-specific markers. RESULTS: Complete AZFc deletions, including BPY2, CDY1 and DAZ genes, were identified in 24 men. The proximal breakpoints were clustered between sY1197 and sY1192, and the distal breakpoints were clustered between sY1054 and sY1125 in all but one of the 24 men. The testicular phenotypes of men with complete AZFc deletion varied from oligozoospermia, to hypospermatogenesis, to maturation arrest. CONCLUSION: We identified a group of infertile men with uniform deletion junctions of AZFc in the Taiwan population. Despite this homogeneous genetic defect in the AZFc region, no clear genotype/phenotype correlation could be demonstrated.
關鍵字
名稱Decreased mRNA transcripts of M-phase promoting factor and its regulators in the testes of infertile men.Lin YM, Teng YN, Chung CL, Tsai WC, Lin YH, Lin JS, Kuo PL.Hum Reprod. Jan;21(1):138-44. Epub 2005 Sep 9.
年度2006
類別期刊論文
摘要BACKGROUND: M-phase promoting factor (MPF), which is comprised of Cyclin B and a catalytic subunit, Cdc2, is a key enzyme required for cells to enter M phase in both mitosis and meiosis. MPF activity is controlled by the stimulatory dephosphorylation of the Cdc25 family and the inhibitory phosphorylation of Wee1. We determined the levels of mRNA transcripts of MPF and its regulators in the testes of infertile men, and evaluated the relationship between the transcript levels and patients testicular phenotypes and sperm retrieval results. METHODS AND RESULTS: The mRNA transcript levels of CDC2, CCNB1, CCNB2, CDC25A, CDC25B, CDC25C and WEE1 in the testes of 37 azoospermic patients were examined by quantitative real-time polymerase chain reaction. Significant decreases in CDC2, CCNB1, CCNB2, CDC25A, CDC25C and WEE1 mRNA transcript levels were detected in patients with spermatogenic failure. CDC2 mRNA transcript levels correlated significantly with those of CCNB1 and CCNB2 mRNA. Significantly higher CDC2, CCNB1, CCNB2, CDC25C and WEE1 mRNA transcript levels were detected in 18 patients with successful sperm retrieval than in 11 patients with failed sperm retrieval. CONCLUSIONS: We suggest that the decreased mRNA transcripts of MPF and its regulators play important roles in human spermatogenesis
關鍵字
名稱Screening of Prader-Willi syndrome and Angelman syndrome in school children with moderate to profound mental retardation in southern Taiwan. Su MT, Teng YN, Kuo PL. Acta Paediatr Taiwan. 2007 Mar-Apr;48(2):73-6.
年度2007
類別期刊論文
摘要BACKGROUND AND PURPOSE: Prader-Willi syndrome (PWS) and Angelman syndrome (AS) are caused by deficiencies of gene expression from paternal or maternal chromosome 15q11-q13, respectively. The study was conducted to estimate the prevalence of PWS and AS in children with moderate to profound mental retardation in Taiwan. METHODS: The screening began with methylation studies in all enrolled cases. If methylation results were positive, Fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) was used to determine whether deletion, uniparental disomy, or imprinting mutation was present. RESULTS: Of 1053 children with moderate to profound mental retardation, we identified three cases of AS (0.28%) and one case of PWS (0.09%). CONCLUSIONS: The prevalence of PWS is lower than AS in school children with moderate to profound mental retardation. The greater number of AS identified than that of PWS is most likely a reflection of more severe mental retardation for AS than for PWS.
關鍵字
名稱Expression of various CDC25B isoforms in human spermatozoa.Teng YN, Chung CL, Lin YM, Pan HA, Liao RW, Kuo PL.Fertil Steril. 2007 Aug;88(2):379-82. Epub 2007 Mar 6.
年度2007
類別期刊論文
摘要OBJECTIVE: To explore the role of CDC25B protein in postmeiotic germ cells. DESIGN: In vitro experiment. SETTING: University-based reproductive genetics laboratory. PATIENT(S): Fertile and infertile volunteers. INTERVENTION(S): Reverse transcription-polymerase chain reaction (RT-PCR), real-time RT-PCR, and immunostaining for CDC25B. MAIN OUTCOME MEASURE: Expression profiling of CDC25B in human spermatozoa. RESULT(S): Four splicing variants (CDC25B1, B2, B3, and B4) are expressed in human spermatozoa. Immunofluorescence staining showed strong homogeneous staining in the midpiece of spermatozoa, and weak staining in the principal piece and cytosol of the head. The messenger RNA (mRNA) transcript level of CDC25B was increased in sperm samples of men with asthenospermia. CONCLUSION(S): The expression of CDC25B in different cellular compartments of human spermatozoa suggests that there are diverse noncell-cycle-related functions of CDC25B in terminally differentiated human germ cells.
關鍵字
名稱A simplified gene-specific screen for Y chromosome deletions in infertile men.Teng YN, Lin YH, Tsai YC, Hsu CC, Kuo PL, Lin YM.Fertil Steril. 2007 Jun;87(6):1291-300. Epub 2007 Feb 12.
年度2007
類別期刊論文
摘要OBJECTIVE: To test the diagnostic efficiency of a gene-specific, five-marker screening strategy for the detection of Y chromosome deletions. DESIGN: Prospective case study. SETTING: University genetics laboratory and reproductive clinics. PATIENT(S): Six hundred twenty-seven infertile men and 212 fertile men. INTERVENTION(S): Peripheral blood samples were screened for Y chromosome deletions in a triple-blind fashion using three protocols: protocol I consisted of five gene-specific markers, including USP9Y, DBY, SMCY, RBM1, and DAZ; protocol II included 14 gene-specific markers; and protocol III consisted of six sequence-tagged sites (STSs) markers recommended by EAA/EMQN. MAIN OUTCOME MEASURE(S): Deletion status of Y chromosome genes or sequence-tagged sites. RESULT(S): Protocols I and II identified the same 41 infertile patients with Y deletions. Protocol III identified 38 infertile patients with Y deletions, and all 38 patients were also identified by protocols I and II. One patient with isolated USP9Y deletion and two patients with isolated DBY deletions, as detected by protocols I and II, could not be identified by protocol III. CONCLUSION(S): We observed mostly consistent results between our protocols and the EAA/EMQN protocol. This gene-specific, five-marker screening panel provides the same diagnostic efficiency as the EAA/EMQN protocol and may be considered an alternative to the EAA/EMQN protocol.
關鍵字
名稱Lee IW, Kuan LC, Lin CH, Pan HA, Hsu CC, Tsai YC, Kuo PL, Teng YN*. Association of USP26 haplotypes in men in Taiwan, China with severe spermatogenic defect. Asian journal of andrology 2008 10(6):896-904.
年度2008
類別期刊論文
摘要AIM: To complete comprehensive haplotype analysis of USP26 for both fertile and infertile men. METHODS: Two hundred infertile men with severe oligospermia or non-obstructive azoospermia were subjected to sequence analysis for the entire coding sequences of the USP26 gene. Two hundred men with proven fertility were genotyped by primer extension methods. Allele/genotype frequencies, linkage disequilibrium (LD) characteristics and haplotypes of fertile men were compared with infertile men. RESULTS: The allele frequencies of five single nucleotide polymorphisms (370-371insACA, 494T>C, 576G>A, ss6202791C>T, 1737G>A) were significantly higher in infertile patients than control subjects. The major haplotypes in infertile men were TACCGA (28% of the population), TGCCGA (15%), TACCAA (8%), TGCCAA (6%), TATCAA (5%) and CATCAA (5%). The major haplotypes for the control subjects were TACCGA (58% of the population), CACCGA (7%), CATCGA (6%) and TGCCGA (5%). Haplotypes TGCCGA, TATCAA, CATCAA, CATCGC, TACCAA and TGCCAA were over-transmitted in patients with spermatogenic defect, whereas haplotypes TACCGA, CACCGA, and CATCGA were under-transmitted in these patients. CONCLUSION: Some USP26 alleles and haplotypes are associated with spermatogenic defect in the Han nationality in Taiwan, China.
關鍵字
名稱Association of extremely skewed X-chromosome inactivation with Taiwanese women presenting with recurrent pregnancy loss.Kuo PL, Huang SC, Chang LW, Lin CH, Tsai WH, Teng YN. J Formos Med Assoc. 2008 Apr;107(4):340-3.
年度2008
類別期刊論文
摘要X-chromosome inactivation (XCI) is a phenomenon that occurs in female mammals. Typically, maternally and paternally-derived X chromosomes are inactivated at approximately the same frequency. If preferential inactivation occurs, the person is considered to have skewed XCI. Skewed XCI has been reported to occur more frequently in women who experience recurrent pregnancy loss (RPL). In this study, we sought to investigate if there is an association between skewed XCI and unexplained RPL in Taiwanese women. A total of 194 women who had experienced unexplained RPL were recruited into the study. Human androgen receptor or DXS6673E and DX15-134 loci were used in the XCI assay. The results of our study suggested that a cut-off point less than 90% may not be justified for skewed XCI. Only extremely skewed (more than 95%) XCI is associated with RPL. Extremely skewed XCI occurs in a subset of Taiwanese women with RPL.
關鍵字
名稱 DAZL protein expression in mouse preimplantation embryo. Pan HA, Liao RW, Chung CL, Teng YN, Lin YM, Kuo PL. Fertil Steril. 2008 May;89(5 Suppl):1324-7. Epub 2007 Aug 30
年度2008
類別期刊論文
摘要Abstract OBJECTIVE: To evaluate the expression pattern of Dazl (deleted in azoospermia-like) protein in the mouse preimplantation embryo. DESIGN: Experimental study. SETTING: Medical research laboratory in a university hospital. ANIMAL(S): Twenty female 28- to 35-day-old FVB mice. INTERVENTION(S): Embryo collection at 1.5, 2.5, and 3.5 days postcoitus (plug date, 0.5 d postcoitus) to examine the Dazl protein expression from the two-cell embryo to the blastocyst. MAIN OUTCOME MEASURE(S): Dazl protein expression was analyzed by immunofluorescent staining. RESULT(S): There is abundant expression of Dazl protein in the cytoplasm of the blastomere. Strong fluorescent signals of Dazl protein expression were found in preimplantation embryo cytoplasm, including two-cell, eight-cell, morula, and blastocyst. CONCLUSION(S): By using an antibody raised against mouse Daz-like protein (Dazl), we showed that Dazl protein is present in all cleaving stages of the preimplantation embryo. This is the first report on the protein expression of a Dazl gene during embryogenesis in mice. However, further study is needed to evaluate the molecular functional role of Dazl
關鍵字
名稱Pan HA, Lee YC, Teng YN, Tsai SJ, Lin YM, Kuo PL. CDC25 protein expression and interaction with DAZL in human corpus luteum. Fertility & Sterility 2009 92(6):1997-2003 (SCI, IF4.167, Ranking 3/61 4.9% in OBSTETRICS & GYNECOLOGY)
年度2009
類別期刊論文
摘要OBJECTIVE: To determine whether the human corpus luteum exhibits CDC25 protein expression and its expression pattern, and identify the interaction with DAZL (deleted in azoospermia-like) in human luteal cells. DESIGN: Experimental study. SETTING: Medical research laboratory in a university hospital. PATIENT(S): Twelve human corpus luteum (CL), four in the early stage, four in the midstage, and four in the late stage of the luteal phase, respectively, and 10 granulosa-lutein cells from IVF patients. INTERVENTION(S): Immunohistochemical stain and Western blot were used to characterize the expression of CDC25 protein, and protein immunoprecipitation was used to identify the interaction of CDC25 with DAZL in human luteal cells. MAIN OUTCOME MEASURE(S): CDC25 protein expression pattern in different stages of luteal phase and interaction with DAZL. RESULT(S): In this study, we show evidence that CDC25 protein is express in granulosa-lutein cells from different stages of CL, and identified only the CDC25A interaction with DAZL in luteal cells. CONCLUSION(S): The CL is the final form of a developing follicle and is the major endocrine component of the ovary in maintaining early successful pregnancy. Expression of CDC25 protein throughout different stages of the ovarian cycles implies important functional roles in the regulation of female reproduction.
關鍵字CDC25, corpus luteum, DAZL, luteal cell
名稱Teng YN*, Kuo PL, Cheng TC, Liao MH. Histone gene expression profile during Spermatogenesis. Fertility & Sterility 2010 93(7):2447-9. (SCI, IF4.167, Ranking 3/61 4.9% in OBSTETRICS & GYNECOLOGY)
年度2010
類別期刊論文
摘要Gene bank search and reverse transcription-polymerase chain reaction were used to analyze the expression profile for histone genes during spermatogenesis. The objective of this study was to provide a systematic view of histone gene expression.
關鍵字histone, spermatogenesis, fertility
名稱Expression of LRWD1 in mouse testis and its centrosomal localization. International Journal of Andrology, 33(6): 832-40. (SCI, IF 3.591, Ranking/Category 1/6 16.66% in ANDROLOGY). NSC 95-2320-B-041-006.
年度2010
類別期刊論文
摘要Abstract The mouse leucine-rich repeats and WD repeat domain containing 1 (lrwd1) gene is located on chromosome 5qG2 and spans over 13 kilobases. It encodes a novel protein of 648-amino acid protein that shares 78.3% amino acid sequence identity with the human LRWD1 protein. We used an oligopeptide as immunogen to generate an anti-lrwd1 antibody in rabbits. Both Northern and Western blot results indicated that the expression of lrwd1 is testis specific. Immunostaining of mouse testis sections detected high levels of lrwd1 signals in the cytoplasm of primary spermatocytes to mature spermatozoa and much weaker signals in spermatogonia. On mature spermatozoa, the anti-lrwd1 antibody stained strongly the connection region between the head and the neck where the centrosome is located. Additional immunostaining and immunoprecipitation showed colocalization and interaction between lrwd1 and γ-tubulin respectively, implicating lrwd1 as a candidate centrosomal protein. These results suggest that lrwd1 may play an important role in spermatogenesis.
關鍵字LRWD1, centrosome, spermatogenesis, testis
名稱Induction of Bcl-2 expression by hepatitis B virus pre-S2 mutant large surface protein resistance to 5-fluorouracil treatment in Huh-7 cells. Hung JH, Teng YN, Wang LH, Su IJ, Wang CC, Huang W, Lee KH, Lu KY, Wang LH. Plos One, 6(12):e28977. (SCI, IF 4.092, Ranking/Category 12/85 14.11% in BIOLOGY). NSC 100-2320-B-041-007.
年度2011
類別期刊論文
摘要PLoS One. 2011;6(12):e28977. Epub 2011 Dec 22. Induction of Bcl-2 expression by hepatitis B virus pre-S2 mutant large surface protein resistance to 5-fluorouracil treatment in Huh-7 cells. Hung JH, Teng YN, Wang LH, Su IJ, Wang CC, Huang W, Lee KH, Lu KY, Wang LH. SourceDepartment of Biotechnology, Chia Nan University of Pharmacy and Science, Tainan, Taiwan. hung86@mail.chna.edu.tw Abstract BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide with poor prognosis due to resistance to conventional chemotherapy and limited efficacy of radiotherapy. Our previous studies have indicated that expression of Hepatitis B virus pre-S2 large mutant surface antigen (HBV pre-S2Δ) is associated with a significant risk of developing HCC. However, the relationship between HBV pre-S2Δ protein and the resistance of chemotherapeutic drug treatment is still unclear. METHODOLOGY/PRINCIPAL FINDINGS: Here, we show that the expression of HBV pre-S2Δ mutant surface protein in Huh-7 cell significantly promoted cell growth and colony formation. Furthermore, HBV pre-S2Δ protein increased both mRNA (2.7±0.5-fold vs. vehicle, p0.05) and protein (3.2±0.3-fold vs. vehicle, p0.01) levels of Bcl-2 in Huh-7 cells. HBV pre-S2Δ protein also enhances Bcl-2 family, Bcl-xL and Mcl-1, expression in Huh-7 cells. Meanwhile, induction of NF-κB p65, ERK, and Akt phosphorylation, and GRP78 expression, an unfolded protein response chaperone, were observed in HBV pre-S2Δ and HBV pre-S-expressing cells. Induction of Bcl-2 expression by HBV pre-S2Δ protein resulted in resistance to 5-fluorouracil treatment in colony formation, caspase-3 assay, and cell apoptosis, and can enhance cell death by co-incubation with Bcl-2 inhibitor. Similarly, transgenic mice showed higher expression of Bcl-2 in liver tissue expressing HBV pre-S2Δ large surface protein in vivo. CONCLUSION/SIGNIFICANCE: Our result demonstrates that HBV pre-S2Δ increased Bcl-2 expression which plays an important role in resistance to 5-fluorouracil-caused cell death. Therefore, these data provide an important chemotherapeutic strategy in HBV pre-S2Δ-associated tumor. c 2011 Hung et al.
關鍵字HBV, Bcl-2, NF-kB
名稱張章堂、莊一全、林宏明、黃珮珊、杜平惪、鄧燕妮∗,運用螢光細胞於環境污染物質的檢測,臺南大學環境與生態學報,第五卷,第一期,頁29-40。
年度2012
類別期刊論文
摘要摘要 本研究運用具有綠色螢光蛋白 (Green Fluorescent Protein, GFP)的小鼠胚胎細胞(mouse embryonic feeder cells ,MEF)暴露在環境污染物檢測中,此螢光蛋白是一種高敏感度的螢光蛋白,實驗結果發現,在僅有50個細胞的情況下,即可偵測得到螢光。相對於一般實驗常用的MTT試驗,則需要1000個細胞以上,方能偵測得到細胞的存在。也就是說,GFP螢光細胞具有極高的敏感度及簡易偵測的優勢,在極低的細胞量就可以被偵測得到,有極高的潛力與優勢適合運用於環境毒物檢測中。以不同毒性物質H2O2、Benzo[a]pyrene(BaP)及來自環境中的柴油廢棄收集液等進行螢光試驗,細胞螢光強度與H2O2及Benzo[a]pyrene(BaP)的處理濃度呈負相關,顯示當毒性物質對於細胞毒殺作用越激烈時,細胞的螢光發散能力將受到影響。未來將可運用於其他領域的生物毒性檢測,包括:化妝品、中草藥、食品等,因此具有極高的外溢效應。此螢光細胞檢測技術平台的建立,不僅可以運用於環境污染物,利用螢光細胞之螢光發散的高敏感性,開發以螢光細胞作為檢測對象的快速檢測平台,藉由此檢測平台的建立,期望未來環境污染物的生物毒性檢測,能達到高敏感度及快速檢測的方向發展。
關鍵字小鼠胚胎細胞、綠色螢光細胞、生物毒性檢測
名稱Endoplasmic Reticulum Stress Stimulates p53 Expression through NF-κB Activation.Lin WC, Chuang YC, Chang YS, Lai MD, Teng YN, Su IJ, Wang CC, Lee KH, Hung JH. Plos One, 7(7):e39120. (SCI, IF 4.092, Ranking/Category 12/85 14.11% in BIOLOGY). NSC 99-2320-B-041-001.
年度2012
類別期刊論文
摘要Abstract BACKGROUND: Induction of apoptosis by endoplasmic reticulum (ER) stress is implicated as the major factor in the development of multiple diseases. ER stress also appears to be a potentially useful major response to many chemotherapeutic drugs and environmental chemical compounds. A previous study has indicated that one major apoptotic regulator, p53, is significantly increased in response to ER stress, and participates in ER stress-induced apoptosis. However, the regulators of p53 expression during ER stress are still not fully understood. PRINCIPAL FINDINGS: In this report, we demonstrate that induction of p53 expression is mediated through NF-κB signaling pathways during ER stress in MCF-7 cells. Tunicamycin or brefeldin A, two ER stress inducers, increased p53 expression in MCF-7 and Hela cells. We found p53 nuclear localization, activity, and phosphorylation at serine 15 on p53 increased during ER stress. Nuclear translocation of NF-κB and activity of NF-κB were also observed during ER stress. ER stress-induced p53 expression was significantly inhibited by coincubation with the NF-κB inhibitor, Bay 11-7082 and downregulation of NF-κB p65 expression. The role of p53 in mediating Brefeldin A-induced apoptosis was also investigated. Induction of p53 expression by Brefeldin A was correlated to Brefeldin A-induced apoptosis. Furthermore, downregulation of p53 expression by p53 siRNA significantly reduced Brefeldin A-induced apoptosis in MCF-7 cells. SIGNIFICANCE: Taken together, NF-κB activation and induction of p53 expression is essential for ER stress-induced cell death which is important for therapeutic effects of clinical cancer drugs. Our results may provide insight into the mechanism of cancer chemotherapy efficacy that is associated with induction of ER stress.
關鍵字NF-kB, p53, Endoplasmic Reticulum Stress
名稱Nuclear factor-κB (NF-κB) Regulates the Expression of Human Testis-Enriched Leucine-rich Repeats and WD repeat Domain containing 1 (LRWD1) Gene, Yen-Ni Teng*, Po-Jung Chuang, and Yo-Wen Liu. International Journal of Molecular Sciences 2012; 14(1):625-39. (SCI, IF 2.598, Ranking/Category 45/154 29.22% in CHEMISTRY, MULTIDISCIPLINARY). NSC 99-2314-B-024-001-MY3.
年度2012
類別期刊論文
摘要The human Leucine-rich Repeats and WD repeat Domain containing 1 (LRWD1) gene was originally identified by cDNA microarray as one of the genes down-regulated in the testicular tissues of patients with severe spermatogenic defects. Human LRWD1 is a testicular-enrich protein that is present predominantly in the cytoplasm of spermatocytes and spermatids and colocalizes with the centrosome at the base of sperm tail. Reporter assay, Chromatin immunoprecipitation (ChIP) analysis, and gel electrophoretic mobility shift assay (EMSA) were used to identify the core promoter region of LRWD1. A 198 bp segment upstream of the LRWD1 transcription initiation site exhibited promoter activity. The LRWD1 core promoter lacked a TATA box but contained a NF-B binding site. Chromatin immunoprecipitation (ChIP) analysis and gel electrophoretic mobility shift assay (EMSA) showed basal binding of the NF-κB subunit to the LRWD1 promoter. LRWD1 promoter activity was positively regulated by NF-κB, and this regulation was dependent on the presence of the conserved κB site in the LRWD1 promoter region. Our data suggest that NF-κB is an important regulator for the expression of LRWD1. This is the first study showing that the expression of the testis-enriched LRWD1 gene is regulated by NF-κB.
關鍵字NF-κB; LRWD1; promoter
名稱A single-nucleotide polymorphism of the DAZL gene promoter confers susceptibility to spermatogenic failure in the Taiwanese Han.Teng YN, Chang YP, Tseng JT, Kuo PH, Lee IW, Lee MS, Kuo PL. Human Reproduction, 27(9):2857-65. (SCI, IF 4.475, Ranking/Category 3/79 3.79% in OBSTETRICS & GYNECOLOGY). NSC 97-2628-B-006-012-MY3.
年度2012
類別期刊論文
摘要Hum Reprod. 2012 Sep;27(9):2857-65. Epub 2012 Jun 29. A single-nucleotide polymorphism of the DAZL gene promoter confers susceptibility to spermatogenic failure in the Taiwanese Han. Teng YN, Chang YP, Tseng JT, Kuo PH, Lee IW, Lee MS, Kuo PL. SourceDepartment of Biological Sciences and Technology, National University of Tainan, Tainan, Taiwan. Abstract BACKGROUND Deleted in AZoospermia-like (DAZL) is an autosomal homologue of Y chromosome-linked DAZ gene located on chromosome 3p24. DAZL is only expressed in the gonads and is critical to germ cell development in different species. However, the regulation of DAZL has not been explored. METHODS Reporter assays, electrophoretic mobility shift assays, supershift assays and bisulfate sequencing were used to identify the core promoter region of DAZL. Sequence analysis was used to identify single-nucleotide polymorphisms (SNPs) in the promoter region. A total of 337 infertile men with abnormal semen parameters and 203 fertile men with normal semen parameters were subjected to sequence analysis of the DAZL promoter region. RESULTS The DAZL gene core promoter is located 1 kb upstream of the transcription start site. Three SNPs (-792G>A, -669A>C and -309T>C) were identified in our population. Of these three SNPs, -792G>A was more prevalent in the infertile men (P 0.0005). Quantitative analysis revealed that genotypes of -792G>A had effects on sperm concentration (P 0.0025) and motility (P 1.5 × 10(-7)). The G to A substitution was associated with decreased binding of the nuclear respiratory factor-1 (NRF-1) to the promoter region and decreased reporter gene activity. CONCLUSION We have identified the core promoter of the human DAZL gene. We also provide preliminary evidence for the role of a novel SNP of the DAZL gene promoter in human spermatogenic failure.
關鍵字single-nucleotide polymorphism / DAZL / gene promoter / spermatogenic failure
名稱SEPT12-microtubule complexes are required for sperm head and tail formation
年度2013
類別期刊論文
摘要SEPT12-microtubule complexes are required for sperm head and tail formation. Kuo PL1, Chiang HS, Wang YY, Kuo YC, Chen MF, Yu IS, Teng YN, Lin SW, Lin YH. The septin gene belongs to a highly conserved family of polymerizing GTP-binding cytoskeletal proteins. SEPTs perform cytoskeletal remodeling, cell polarity, mitosis, and vesicle trafficking by interacting with various cytoskeletons. Our previous studies have indicated that SEPTIN12+/+/+/- chimeras with a SEPTIN12 mutant allele were infertile. Spermatozoa from the vas deferens of chimeric mice indicated an abnormal sperm morphology, decreased sperm count, and immotile sperm. Mutations and genetic variants of SEPTIN12 in infertility cases also caused oligozoospermia and teratozoospermia. We suggest that a loss of SEPT12 affects the biological function of microtublin functions and causes spermiogenesis defects. In the cell model, SEPT12 interacts with α- and β-tubulins by co-immunoprecipitation (co-IP). To determine the precise localization and interactions between SEPT12 and α- and β-tubulins in vivo, we created SEPTIN12-transgene mice. We demonstrate how SEPT12 interacts and co-localizes with α- and β-tubulins during spermiogenesis in these mice. By using shRNA, the loss of SEPT12 transcripts disrupts α- and β-tubulin organization. In addition, losing or decreasing SEPT12 disturbs the morphogenesis of sperm heads and the elongation of sperm tails, the steps of which are coordinated and constructed by α- and β-tubulins, in SEPTIN12+/+/+/- chimeras. In this study, we discovered that the SEPTIN12-microtubule complexes are critical for sperm formation during spermiogenesis.
關鍵字septin 12
名稱Correlation between leucine rich domain and the stability of LRWD1 protein in human NT2/D1 cells
年度2014
類別期刊論文
摘要PURPOSE: LRWD1 is a protein that contains LRR and WDs domains and is important in regulating spermatogenesis. However, the roles of LRR or WDs domains in the expression of LRWD1 remain unclear. MATERIALS AND METHODS: The NT2/D1 cells separately transfected with full length of LRWD1 gene (LRWD(WT)) or genes with deleted sequences in the LRR domain (LRWD1(ΔLRR)), WD1 domain (LRWD1(ΔWD1)), WD2 domain (LRWD1(ΔWD2)), WD3 domain (LRWD1(ΔWD3)) and entire three WD domains (LRWD1(Δ3×WD)) were applied to investigate the expression levels of LRWD1 protein by either Western blot or flow cytometry. The associated proteins in these mutated LRWD1 proteins were identified by mass spectrometry. RESULTS: Deletion of the LRR domain significantly decreased the expression of LRWD1 protein. With the treatment of MG132, the LRR domain may functions in preventing LRWD1 protein from proteasome-mediated degradation. In the co-immunoprecipitation analysis, protein receptor of tumor necrosis factor 2 (TNFR2) was specifically observed to be associated with LRR-deficient LRWD1 protein. CONCLUSIONS: The LRR domain is significantly correlated to the stability of LRWD1 protein. Determining if the stability is modulated by TNFR2 is worthy of further study.
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名稱The antiproliferative and apoptotic effects of sirtinol, a sirtuin inhibitor on human lung cancer cells by modulating Akt/β-catenin-Foxo3a axis
年度2014
類別期刊論文
摘要Sirtuins, NAD(+)-dependent deacetylases, could target both histones and nonhistone proteins in mammalian cells. Sirt1 is the major sirtuin and has been shown to involve various cellular processes, including antiapoptosis, cellular senescence. Sirt1 was reported to be overexpressed in many cancers, including lung cancer. Sirtinol, a specific inhibitor of Sirt1, has been shown to induce apoptosis of cancer cells by elevating endogenous level of reactive oxygen species. In the study, we investigated the effect of sirtinol on the proliferation and apoptosis of nonsmall cell lung cancer (NSCLC) H1299 cells. The results of proliferation assay and colony formation assay showed the antigrowth effect of sirtinol. The annexin-V staining further confirmed the apoptosis induction by sirtinol treatment. Interestingly, the levels of phosphorylated Akt and β-catenin were significantly downregulated with treating the apoptotic inducing doses. On the contrary, sirtinol treatment causes the significantly increased level of FoxO3a, a proapoptotic transcription factor targeted by Sirt1. These above results suggested that sirtinol may inhibit cell proliferation of H1299 cells by regulating the axis of Akt-β-catenin-FoxO3a. Overall, this study demonstrates that sirtinol attenuates the proliferation and induces apoptosis of NSCLC cells, indicating the potential treatment against NSCLC cells by inhibiting Sirt1 in future applications.
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名稱Reactive oxygen species mediate Terbufos-induced apoptosis in mouse testicular cell lines via the modulation of cell cycle and pro-apoptotic proteins
年度2015
類別期刊論文
摘要Terbufos (S-t-butylthiomethyl-O,O-diethyl phosphorodithioate) is a highly toxic organophosphate which is extensively used as an insecticide and nematicide. Chronic exposure to terbufos causes neuronal injury and predisposes to neurodegenerative diseases. Accumulating evidence has shown that the exposure to terbufos, as an occupational risk factor, may also cause reproductive disorders. However, the exact mechanisms of reproductive toxicity remain unclear. The present study aimed to investigate the toxic effect of terbufos on testicular cells and to explore the mechanism of toxicity on a cellular level. The cytotoxic effects of terbufos on mouse immortalized spermatogonia (GC-1), spermatocytes (GC-2), Leydig (TM3), and Sertoli (TM4) cell lines were assessed by MTT assays, caspase activation, flow cytometry, TUNEL assay, Western blot, and cell cycle analysis. The exposure to different concentrations of terbufos ranging from 50 to 800 μM for 6 h caused significant death in all the used testicular cell lines. Terbufos increased reactive oxygen species (ROS) production, reduced mitochondrial membrane potential, and initiated apoptosis, which was confirmed by a dose-dependent increase in the number of TUNEL-positive apoptotic cells. Blocking ROS production by N-acetyl cysteine (NAC) protected GC-1 cells from terbufos-induced cell death. The results demonstrated that terbufos induces ROS, apoptosis, and DNA damage in testicular cell lines and it should be considered potentially hazardous to testis. Together, this study provided potential molecular mechanisms of terbufos-induced toxicity in testicular cells and suggests a possible protective measure. © 2015 Wiley Periodicals, Inc. Environ Toxicol, 2015.
關鍵字terbufos, mouse testicular cell lines, apoptosis, caspase-3, reactive oxygen species.
名稱6-Shogaol induces cell cycle arrest and apoptosis in human hepatoma cells through pleiotropic mechanisms
年度2015
類別期刊論文
摘要Abstract Shogaols are a group of the active constituents of ginger that have been identified to have various biological activities. The aim of the current study was to investigate the antitumor activity of 6-shogaol in hepatocellular carcinoma (HCC) and the possible involvement of reactive oxygen species as a putative mechanism of action. HCC cell lines, HepG2 and Huh-7, were used to study the in vitro anti-cancer activity of 6-shogaol via the application of various molecular biology techniques. Results showed that 6-shogaol effectively inhibited the cell viability, caused cell cycle arrest at G2/M phase and induced apoptosis in HCC cells as indicated by MTT assay, DAPI nuclear staining, annexin V assay, cell cycle analysis, and activation of caspase-3. Western blot analysis revealed the ability of 6-shogaol to target cancer survival signaling pathways mediated by mitogen-activated protein kinase (MAPK), 5 AMP-activated protein kinase (AMPK) and Akt. In addition, 6-Shogaol induced alteration of cyclin proteins expression and caused cleavage of protein kinase C delta. Furthermore, 6-Shogaol was able to induce the production of reactive oxygen species and endoplasmic reticulum (ER) stress-associated proteins and the consequent activation of autophagy in HepG2 cells. Taken together, the current study highlights evidences that 6-shogaol induces apoptosis, modulates cyclins expression and targets cancer survival signaling pathways in HCC cell lines, at least in part, via the production of reactive oxygen species. These findings support 6-shogaols clinical promise as a potential candidate for HCC therapy. Copyright © 2015 Elsevier B.V. All rights reserved.
關鍵字6-Shogaol; Apoptosis; Autophagy; Cell cycle arrest; Endoplasmic reticulum stress; Mitogen-activated protein; Reactive oxygen species
名稱Inhibitory effect of trans-ferulic acid on proliferation and migration of human lung cancer cells accompanied with increased endogenous reactive oxygen species and β-catenin instability.
年度2016
類別期刊論文
摘要BACKGROUND: Trans-ferulic (FA) acid exhibits antioxidant effects in vitro. However, the underlying mechanism of trans-FA activity in cellular physiology, especially cancer physiology, remains largely unknown. This study investigated the cellular physiological effects of trans-FA on the H1299 human lung cancer cell line. METHODS: The 2,2-diphenyl-1-picrylhydrazyl assay was used to determine free radical scavenging capability. Assessment of intracellular reactive oxygen species (ROS) was evaluated using oxidized 2,7-dichlorofluorescin diacetate and dihydroethidium staining. Trypan blue exclusion, colony formation, and anchorage-independent growth assays were used to determine cellular proliferation. Annexin V staining assay was used to assess cellular apoptosis by flow cytometry. Wound healing and Boydens well assays were used to detect the migration and invasion of cells. Gelatin zymography was used to detect matrix metalloproteinase (MMP-2 and MMP-9) activity. Western blotting was used to detect expression levels of various signaling pathway proteins. RESULTS: DPPH assay results indicated that trans-FA exerted potent antioxidant effects. However, trans-FA increased intracellular ROS levels, including hydrogen peroxide and superoxide anion, in H1299 cells. Trans-FA treatment inhibited cellular proliferation and induced moderate apoptotic cell death at the highest concentration used (0.6 mM). Furthermore, trans-FA moderately inhibited the migration of H1299 cells at the concentrations of 0.3 and 0.6 mM and attenuated MMP-2 and MMP-9 activity. Trans-FA caused the phosphorylation of β-catenin, resulting in proteasomal degradation of β-catenin. Conversely, trans-FA treatment increased the expression of pro-apoptotic factor Bax and decreased the expression of pro-survival factor survivin. CONCLUSION: Various concentrations (0.06-0.6 mM) of trans-FA exert both anti-proliferation and anti-migration effects in the human lung cancer cell line H1299.
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名稱The adverse effects of low-dose exposure to Di(2-ethylhexyl) phthalate during adolescence on sperm function in adult rats
年度2016
類別期刊論文
摘要Di(2-ethylhexyl) phthalate (DEHP) is the most crucial phthalate derivative added to polyvinyl chloride as a plasticizer. This study examined the effects of low-dose exposure to DEHP during adolescence on sperm function in adult rats. The male rats were daily gavaged with 30, 100, 300, and 1000 μg kg-1 of DEHP or corn oil from postnatal day (PND) 42 until PND 105. The selection of DEHP doses ranged from the mean daily intake by the normal-population exposure levels to no-observed-adverse-effect level of DEHP for the endpoints evaluated until adulthood. Significant increases in the percentage of sperm with tail abnormality, tendency for sperm DNA fragmentation index (DFI) and percentage of sperm with DFI were found in those exposed to 100, 300, and 1000 μg kg-1 (P??0.05). We observed a significant increase of hydrogen peroxide (H2 O2 ) generation in the sperm of the 1000 μg kg-1 group compared with the control group (P??0.05). The excessive production of sperm H2 O2 coincided with an increase in sperm DFI. In this study, the lowest-observed-adverse-effect level for sperm toxicity was considered to be 100 μg DEHP/kg/day in sperm morphology and chromatin DNA damage. Further research is necessary to clarify the mechanisms of DEHP-related sperm ROS generation on sperm DNA damage. c 2014 Wiley Periodicals, Inc. Environ Toxicol, 2014.
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名稱Effects of 2,3′,4,4′,5-pentachlorobiphenyl (PCB 118) in SD rats testicular toxicity in F3 offspring.
年度2012
類別研討會
摘要
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名稱Human LRWD1 Is Associated with Centrosomes and Microtubules Organization
年度2013
類別研討會
摘要 LRWD1 (Leucine-Rich repeats and WD repeat domain containing 1) is highly expressed in the testes, but the roles in male germ cell development and spermatogenesis remain unclear. The objective of this study is to investigate the expression and function of LRWD1 in human sperms. We studied the expression of LRWD1 in fertile males and infertile patients by RT-PCR and immunostaining. LRWD1 was detected in the testes of all human subjects except Sertoli-cell-only-syndrome (SCOS) patients. In spermatozoa, LRWD1 was found to colocalized with centrin and γ-tubulin in the centrosome region. We also applied the immunofluorescence assay (IFA) to study the expression of LRWD1 in ejaculated spermatozoa of various patients with sperm abnormality. In the neck- and tail-defective groups, we found that significant reduction of sperms LRWD1 immunostaining signals. Sperm tails is the structure extending from centrosome. LRWD1 localizes at the centrosome in human spermatozoa and correlates with multiple sperm morphological defects according to the decrease of LRWD1. We hypothesized that human LRWD1 is a centrosomal protein and plays an important role for the microtubule organization in the cell cytoskeleton and sperm tails formation. Knockdown of LRWD1 in HeLa cells by siRNA demonstrated that LRWD1 participates in microtubule organization and function. In IFA experiment, the LRWD1 colocalize with γ-tubulin in the centrosome region. Rescue after vinblastine treatment, LRWD1 have more signal on the centrosome. We suggest that LRWD1 may have important role in cell cytoskeleton formation.
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名稱Leucine-Rich Repeats and WD Repeat Domain Containing 1 (LRWD1) is associated with Reactive oxygen species (ROS) response in testicular cells.
年度2014
類別研討會
摘要
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名稱Hypermethylation of the Leucine-Rich Repeats and WD Repeat Domain Containing 1 (LRWD1) Promoter in sperm.
年度2014
類別研討會
摘要
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名稱Transcriptional regulation and Expression of Human Leucine-Rich Repeats and WD Repeat Domain Containing 1 (LRWD1).
年度2015
類別研討會
摘要Study Question: Leucine-rich repeats and WD repeat domain containing protein 1 (LRWD1) is richly expressed in the testes, yet its roles and transcriptional regulation in human male germ cell development remain unclear. The objective of this study is to investigate the expression and function of LRWD1 in human sperms and the transcriptional regulation in human sperms. Materials, Setting, Methods: We studied the expression of LRWD1 in sperm s of fertile males and infertile patients by RT-PCR and immunostaining. In addition, we further compared the expression of LRWD1 in the mature spermatozoa from normal controls and from patients with morphologically defective sperms. To assess whether the DNA methylation play an important role in transcriptional regulation of LRWD1, we analyzed the transcriptional activity and revealed the CpG islands of immediately upstream of LRWD1. Main Results: In mature spermatozoa, we detected LRWD1 in the centrosome close to the connection region between the head and neck. The fraction of sperm with reduced level of LRWD1 was significantly increased in the semen samples of patients with asthenozoospermia, teratozoospermia, and asthenoteratozoospermia. Sperms without the LRWD1 signal were increased in the neck-defective and tail-defective groups. In addition, LRWD1 was highly expressed in the testes, and associated with the sperm morphology and motility. In the quantitative methylation-specific PCR (qMSP) showed the DNA hypermethylation of LRWD1 promoter was negatively correlated to sperm motility. Altogether, LRWD1expression is mediated by methylation in human sperms.
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名稱Reactive Oxygen Species (ROS) and miR-320a regulated LRWD1 expression
年度2016
類別研討會
摘要Study Question: LRWD1 (Leucine-Rich repeats and WD repeat domain containing 1) was highly expressed in the testes, and associated with the sperm morphology and motility. ROS are free radicals that are involved in numerous sperm physiological processes such as capacitation, hyper activation, and sperm-oocyte fusion. The objective of this study was to investigate the association between the reactive oxygen species and LRWD1 expression in testicular cell of human. Study Design, Size, Duration: To assess whether the ROS play important role in LRWD1 of testicular cell human, we used hydrogen peroxide (H2O2) or sodium nitroprusside dehydrate (SNP) treatment in testicular cell, and investigated the role of miRNA for LRWD1 expression. Materials, Setting, Methods: Hydrogen peroxide (H2O2) or Sodium nitroprusside dehydrate (SNP) treatment to testicular cell to NT2D1 cell ,and then assay LRWD1 expression by western blot and real-time PCR. miR-320a have potential bind site on the 3’UTR of LRWD1 transcript by the microRNA analyzing tools, miRanda, TargetScan,Microcosm, StarBase and RNAhybrid. In the study, we reported the effects of LRWD1 on reactive oxygen species and miR-320a. We made LRWD1-3’UTR fused to a luciferase reporter vector to construct pMIR-LRWD1-3’UTR. Main Results: The expression of miR-320a and LRWD1 were increased by hydrogen peroxide (H2O2) or sodium nitroprusside dehydrate (SNP) treatment in TaqMan real time-PCR assay. Conclusion: With this study, the post-transcriptional regulation of miR-320a for LRWD1 help to understand the function and roles of the miRNA in post-transcriptional regulation of LRWD1 and may provide a rationale for further diagnosis and treatment of spermatogenic defect and male infertility diseases. Acknowledgment This study was supported by grants from National Science Council (NSC 102-2314-B-024 -001) and Ministry of Science and Technology (MOST 103-2314-B-024 -002, MOST 104-2314-B-024 -002 -MY2) of Taiwan.
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