Decreased expression of SAM68 in human testes with spermatogenic defects

作者:Li, Le-Jun; Zhang, Feng-Bin; Liu, Shu-Yuan; Tian, Yong-Hong; Le, Fang; Lou, Hang-Ying; Huang, He-Feng; Jin, Fan*
来源:Fertility and Sterility, 2014, 102(1): 61-+.
DOI:10.1016/j.fertnstert.2014.03.036

摘要

Objective: To assess the expression patterns of SAM68 in the testes of azoospermic patients with normal and abnormal spermatogenesis. @@@ Design: Retrospective study and in vitro study. @@@ Setting: University hospital. @@@ Patient(s): 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). @@@ Intervention(s): No interventions with patients. Knockdown of Sam68 was performed in the GC-2spd(ts) cell line. @@@ Main Outcome Measure(s): 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. @@@ Result(s): 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. @@@ Conclusion(s): 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.