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原癌基因{sl c--src}通过磷酸化信号转导与转录激活子--3蛋白影响大鼠精原干细胞活性

陈加祥, 王新长, 王晶磊, 徐斯凡, 秦海燕, 杨蓓, 杨俊玲, 邹挺

南昌大学医学院生理学教研室.江西,南昌 330006；中央民族大学生命与环境科学学院.北京 100081

摘要

该文旨在研究原癌基因{sl c--src}在体外培养的9日龄大鼠精原干细胞中的表达及其与精原干细胞活性相关的信号通路。用MTT比色法观察反义{sl c--src}脱氧寡核苷酸(oligodeoxynucleotides, ODNs)对精原干细胞作用的量效及时效关系；用RT--PCR检测{sl c--src} mRNA的表达；用Western blot检测{sl c--src}表达产物PP60c--src和磷酸化的信号转导与转录激活子--3 (phosphorylated signal transducer and activator of transcription--3, p--STAT3)的表达。结果显示，与空白对照组相比，10 #mu#mol/L反义{sl c--src} ODNs作用12 h后精原干细胞活性下降8.1% ({sl P}<0.05)，且{sl c--src} mRNA表达明显下调；Western blot结果显示反义{sl c--src} ODNs组PP60c--src、p--STAT3蛋白表达与空白对照组相比分别下降了33.8%、45.3% (均{sl P}<0.01)。原癌基因{sl c--src}可能通过p--STAT3蛋白影响精原干细胞的活性。

关键词： 大鼠; 精原干细胞; 原癌基因c-src; 信号转导与转录激活子-3

Proto--oncogene {sl c--src} regulates the viability of rat spermatogonial stem cells {sl in vitro} through phosphorylated signal transducer and activator of transcription--3

Chen Jiaxiang, Wang Xinchang, Wang Jinglei, Xu Sifan, Qin Haiyan, Yang Bei, Yang Junling, Zou Ting

Department of Physiology,Medical College of Nanchang University.Nanchang 330006,Jiangxi；China

Abstract

The present study aimed to investigate the effect of proto-oncogene c-src on the viability of rat spermatogonial stem cells in 9 day-old rat. MTT method was used to observe the viability of the spermatogonial stem cells treated with antisense src oligodeoxynucleotides (ODNs) in vitro; RT-PCR was utilized to observe the expressions of c-src mRNA and Western blot was used to observe the PP60c-src and phosphorylated signal transducer and activator of transcription-3 (p-STAT3) contents. Compared with that in control group, the viability of spermatogonial stem cells decreased by 8.1% (P<0.05) and the expression of c-src mRNA decreased significantly after being treated with 10 #mu#mol/L antisense src ODNs for 12 h. Compared with the control group, the PP60c-src and p-STAT3 contents decreased by 33.8% and 45.3% (both P<0.01), respectively, in the spermatogonial stem cells after transfected antisense c-src ODNs. The results suggest that proto-oncogene c-src regulates the viability of rat spermatogonia stem cells through p-STAT3.

Key words: Rat;spermatogonial stem cells;proto-oncogene c-src;signal transducer and activator of transcription-3

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引用本文：

陈加祥, 王新长, 王晶磊, 徐斯凡, 秦海燕, 杨蓓, 杨俊玲, 邹挺. 原癌基因{sl c--src}通过磷酸化信号转导与转录激活子--3蛋白影响大鼠精原干细胞活性[J]. 生理学报 2008; 60 (3): 391-396.

Chen Jiaxiang, Wang Xinchang, Wang Jinglei, Xu Sifan, Qin Haiyan, Yang Bei, Yang Junling, Zou Ting. Proto--oncogene {sl c--src} regulates the viability of rat spermatogonial stem cells {sl in vitro} through phosphorylated signal transducer and activator of transcription--3. Acta Physiol Sin 2008; 60 (3): 391-396 (in Chinese with English abstract).