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SAM68 在生精缺陷的人睾丸中表达降低。

Decreased expression of SAM68 in human testes with spermatogenic defects.

机构信息

Department of Reproductive Endocrinology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, People's Republic of China.

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.

出版信息

Fertil Steril. 2014 Jul;102(1):61-67.e3. doi: 10.1016/j.fertnstert.2014.03.036. Epub 2014 Apr 29.

Abstract

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.

摘要

目的

评估 SAM68 在正常和异常生精的无精子症患者睾丸中的表达模式。

设计

回顾性研究和体外研究。

设置

大学医院。

患者

正常生精的无精子症患者(OAZ;n=20)、精母细胞阶段阻滞的无精子症患者(MA;n=20)和唯支持细胞综合征(SCOS;n=10)的睾丸活检。

干预措施

对患者无干预。在 GC-2spd(ts)细胞系中敲低 Sam68。

主要观察指标

使用定量实时逆转录聚合酶链反应(RT-PCR)、Western blot 和免疫组织化学分析在组织中分析 SAM68 表达。此外,在 GC-2spd(ts)细胞中敲低 Sam68。使用 MTT 测定法测量细胞活力,并使用 Annexin V-FITC 试剂盒通过流式细胞术检测细胞凋亡率。

结果

使用 qRT-PCR,与 OAZ 患者相比,MA 和 SCOS 患者睾丸中 SAM68 mRNA 的表达水平明显降低。此外,使用 qRT-PCR、Western blot 和免疫组织化学分析,在 45%(9/20)的 MA 患者和所有 SCOS 患者的睾丸组织中,SAM68 的 mRNA 和蛋白表达缺失或几乎无法检测到。此外,Sam68 表达下调抑制了转染的 GC-2spd(ts)细胞的增殖并诱导其凋亡。

结论

在精母细胞阶段阻滞的无精子症和 SCOS 的人类睾丸中观察到 SAM68 表达不足。这些结果可能为异常生精的分子基础提供新的视角。

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