原癌基因A-myb对大鼠精原干细胞活性的影响  

Proto-oncogene A-myb Regulates Viability of Rat Spermatogonial Stem Cells in Vitro

作　　者：陈加祥[1] 徐林林[2] 王晶磊[1] 

机构地区：[1]南昌大学医学院生理教研室,南昌330006 [2]南昌大学第一附属医院医学科研中心,南昌330006

出　　处：《南昌大学学报（医学版）》2011年第8期52-56,59,共6页Journal of Nanchang University：Medical Sciences

基　　金：国家自然科学基金(No.30360032);江西省教育厅课题(GJJ09444;GJJ09111);南昌大学博士科研启动基金和南昌大学(医科类)基金

摘　　要：目的研究原癌基因A-myb在体外培养的9日龄大鼠精原干细胞活性中的作用。方法用不连续密度的Percoll液结合差速贴壁方法分离和纯化精原干细胞;用RT-PCR检测A-myb mRNA的表达;用Western blot检测A-myb表达产物A-myb的表达;用MTT比色法观察反义A-myb脱氧寡核苷酸(oligodeoxynucleotides,ODNs)对精原干细胞作用的量效及时效关系。结果分离提取的细胞基本上为精原干细胞。与空白对照组相比,10μmol.L-1反义A-myb ODNs作用48h后精原干细胞内A-myb mRNA表达下降了47.8%(P<0.05),A-myb蛋白下降了51.7%(P<0.05)。5、10、20μmol.L-1反义A-myb ODNs组与0μmol.L-1组在作用细胞48h后相比,精原干细胞的活性分别下降11.8%(P<0.05)、50.1%(P<0.01)、53.8%(P<0.01)。与空白对照组相比,10μmol.L-1反义A-myb ODNs组作用12、24、48、72h后精原干细胞活性分别下降18.8%(P<0.05)、30.0%(P<0.01)、38.6%(P<0.01)、41.0%(P<0.01),而正义A-myb ODNs组和空白对照组相比无明显差异。结论原癌基因A-myb可调节大鼠精原干细胞的活性。Objective To investigate the effect of proto-oncogene A-myb on the viability of rat spermatogonial stem cells in 9 day-old rats.Methods The rat spermatogonial stem cells were isolated and purified by percoll discontinuous density gradient centrifugation combined with removal contaminating somatic cells through adhesion to plastic dishes.The expression of A-myb mRNA and protein was detected by RT-PCR and Western blot.The viability of spermatogonial stem cells treated with antisense A-myb oligodeoxynucleotides（ODNs） was observed by MTT method.Results The isolated cells were almost spermatogonial stem cells.Compared with the control,treatment with 10 μmol·L-1 antisense A-myb ODNs for 48 hour decreased the levels of A-myb mRNA and protein by 47.8% and 51.7%,respectively（both P0.05）.Treatment with 5,10 and 20 μmol·L-1 antisense A-myb ODNs decreased the viability of spermatogonial stem cells by 11.8%,50.1% and 53.8%,respectively（P0.05 orP0.01）.Treatment with 10 μmol·L-1 antisense A-myb ODNs for 12,24,48 and 72 hours decreased cell viability by 18.8%,30.0%,38.6% and 41.0%,respectively（P0.05 or P0.01）.Conclusion Proto-oncogene A-myb may regulate the viability of rat spermatogonial stem cells.

关 键 词：精原干细胞 原癌基因A-myb 动物 实验 大鼠 

分 类 号：Q492[生物学—生理学]