野生型PTEN诱导卵巢癌SKOV3细胞生物特性改变  

Wild-type PTEN Induced Changes in Biological Characteristics of Ovarian Cancer SKOV3 Cells

作　　者：唐小龙[1] 何敏[1] 江振友[2] 庞雪云[1] 张荣波[3] 汪雪峰[3] 

机构地区：[1]广东省中医院检验科,广东广州510120 [2]暨南大学医学院,广东广州510120 [3]安徽理工大学医学院生物工程研究所,安徽淮南232001

出　　处：《中山大学学报（医学科学版）》2010年第5期602-607,613,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(30973693)

摘　　要：【目的】探索野生型PTEN基因对卵巢癌细胞生物学特性的影响。【方法】将野生型PTEN基因克隆到pAdxsi腺病毒载体并感染SKOV3细胞。荧光镜下检测腺病毒载体对SKOV3细胞的感染效率;用CCK-8法检测细胞增殖的抑制效率;RT-PCR与WB分别检测PTEN mRNA与蛋白表达水平变化;免疫化学法与间接荧光法检测PTEN在细胞内的定位及细胞内NEDD4-1和RAD51等分子表达。【结果】稳定转染PTEN后,24h内PTEN分布于细胞质与核内,而72h后则主要集中于细胞核内;显著增加细胞质内NEDD4-1和细核RAD51等分子表达。【结论】野生型PTEN可影响SKOV3细胞增殖,并诱导细胞表达RAD51与NEDD4-1分子,有利于细胞DNA修复与细胞增殖抑制。Objective To explore the effect of wild-type PTEN gene on biological characteristics of ovarian cancer cells.Methods The wild-type PTEN gene was cloned into pAdxsi adenovirus vector and infected into SKOV3 cells,the infected efficiency of adenovirus on SKOV3 cells was detected by fluorescence microscopy;the inhibition efficiency on cell proliferation was assayed by CCK-8;RT-PCR and WB was used to detect the level of PTEN mRNA and protein expression changes respectively;the location of PTEN NEDD4-1,RAD51 and other molecules in cell were showed by immunochemical and indirect fluorescence.Results The PTEN could be observed in the cytoplasm and nucleus after 24 h of stable transfection,while after 72 h they could be presented mainly in nucleus;the transfection of PTEN could increase the expression of NEDD4-1,nuclear RAD51 molecules significantly in the cytoplasm.Conclusion The wild-type PTEN can effect the proliferation of SKOV3 cells and induce expression of RAD51 and NEDD4-1 molecules,thus would contribute to cellular DNA repair and cell proliferation inhibition.

关 键 词：转染 NEDD4-1 PTEN基因 

分 类 号：R737.31[医药卫生—肿瘤]