title: Decreased expression of SAM68 in human testes with spermatogenic defects.
abstract: OBJECTIVE
  To assess the expression patterns of SAM68 in the testes of azoospermic patients with normal and abnormal spermatogenesis.
  
  
  METHODS
  Retrospective study and in vitro study.
  
  
  METHODS
  University hospital.
  
  
  METHODS
  Testicular biopsies of azoospermic men with normal spermatogenesis (OAZ; n=20), with maturation arrest at the spermatocyte stage (MA; n=20), and with Sertoli cell-only syndrome (SCOS; n=10).
  
  
  METHODS
  No interventions with patients. Knockdown of Sam68 was performed in the GC-2spd(ts) cell line.
  
  
  METHODS
  SAM68 expression was analyzed using quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot, and immunohistochemistry analysis in tissues. Moreover, Sam68 was knocked down in GC-2spd(ts) cells. Cell viability was measured using the MTT assay, and the apoptosis rate was detected using flow cytometry with the Annexin V-FITC kit.
  
  
  RESULTS
  Using qRT-PCR, the expression level of testicular SAM68 mRNA in MA and SCOS patients was statistically reduced compared with in OAZ patients. In addition, using qRT-PCR, Western blot, and immunohistochemistry analyses, mRNA and protein expressions of SAM68 were absent or barely detectable in testicular tissues in 45% (9 of 20) of patients with MA and in all patients with SCOS. Furthermore, decreased expression of Sam68 suppressed germ cell proliferation and induced apoptosis in transfected GC-2spd(ts) cells.
  
  
  CONCLUSIONS
  Deficient SAM68 expression was observed in the human testis with MA at the spermatocyte stage and SCOS. These results may offer new perspectives on the molecular basis of abnormal spermatogenesis.
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Le-Jun
authors.initials: LJ
authors.lastname: Li
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Feng-Bin
authors.initials: FB
authors.lastname: Zhang
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Shu-Yuan
authors.initials: SY
authors.lastname: Liu
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Yong-Hong
authors.initials: YH
authors.lastname: Tian
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Fang
authors.initials: F
authors.lastname: Le
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Hang-Ying
authors.initials: HY
authors.lastname: Lou
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: He-Feng
authors.initials: HF
authors.lastname: Huang
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, People's Republic of China; Key Laboratory of Reproductive Genetics, National Ministry of Education (Zhejiang University), Women's Reproductive Health Laboratory of Zhejiang Province, Hangzhou, Zhejiang, People's Republic of China. Electronic address: jinfan@zju.edu.cn.
authors.forename: Fan
authors.initials: F
authors.lastname: Jin
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000328:Adult; D000818:Animals; D017209:Apoptosis; D053713:Azoospermia / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation; D006801:Humans; D008297:Male; D051379:Mice; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D012189:Retrospective Studies; D054331:Sertoli Cell-Only Syndrome / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D013091:Spermatogenesis; D013094:Spermatozoa / Q000378:metabolism* / Q000473:pathology; D013737:Testis / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D014162:Transfection
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D004268:DNA-Binding Proteins; C489559:KHDRBS1 protein, human; C513253:Khdrbs1 protein, mouse; D012333:RNA, Messenger; D016601:RNA-Binding Proteins
keywords: RNA-binding proteins; SAM68; male infertility; spermatogenic defect