信号通路抑制剂对PTEN基因不同表达状态子宫内膜癌细胞的作用  

Effect of signal transduction inhibitors on human endometrial carcinoma cells with differential PTEN gene expression

作　　者：肖兰[1] 杨越波[1] 沈慧敏[1] 许成芳[1] 王小韵[1] 李小毛[1] 

机构地区：[1]中山大学附属第三医院妇科,广州510630

出　　处：《中华妇产科杂志》2009年第9期681-685,共5页Chinese Journal of Obstetrics and Gynecology

基　　金：国家自然科学基金（30772332）;广东省自然科学基金（7001568）;广东省科技计划项目（2007B030502014）;广州市科技计划项目（200723-E0601）

摘　　要：目的比较相关信号通路抑制剂诱导PTEN基因不同表达状态子宫内膜癌细胞的增殖和凋亡情况。方法以PTEN反义寡核苷酸及PTEN真核表达载体转染子宫内膜癌细胞（HEC-1A、Ishikawa）后，激光共聚焦显微镜观察PTEN蛋白的表达。表皮生长因子受体、丝裂原活化蛋白激酶及雷帕霉素靶蛋白信号通路抑制剂——RG-14620、SB203580（SB）及西罗莫司分别处理HEC-1A、HEC-1A—PTEN-null、Ishikawa、Ishikawa-PTEN细胞，荧光染色法、流式细胞仪、四甲基偶氮唑蓝比色法分别检测细胞凋亡时的形态学改变、细胞凋亡率、细胞周期分布、细胞存活率。结果转染后，Ishikawa—PTEN细胞内PTEN蛋白表达增加，HEC-1A—PTEN—null细胞内PTEN蛋白表达受到抑制。RG-14620、SB及西罗莫司处理后，Ishikawa和HEC-1A—PTEN—null细胞荧光染色见核呈浓集固缩改变；细胞发生明显凋亡及G1期阻滞，尤以SB作用最为显著，lshikawa＋SB、HEC-1A—PTEN—null＋SB的细胞凋亡率分别为（37．8±0．8）％、（31．6±0．8）％，G1期细胞比例增加[分别为（87．5±1．9）％、（84．1±3．2）％]；细胞增殖受到抑制，尤以SB处理后作用为屁著，Ishikawa＋SB、HEC-1A-PTEN—null＋SB的细胞存活率仅为（49．0±1．7）％、（54．0±2．1）％。结论PTEN基因缺失使相应信号通路抑制剂作用的子宫内膜癌细胞增殖明显受抑，细胞阻滞于G1期，细胞凋亡增加，对相关信号通路抑制剂的敏感性增加。Objective To investigate the apoptotic and proliferation effects of signal transduction inhibitors on human endometrial carcinoma cells with different PTEN gene expression. Methods PTEN antisense oligonucleotide and pcDNA3.1/PTEN vector contained PTEN gene were transfected into endometrial carcinoma cells （HEC-1A and Ishikawa）. The expression of PTEN protein was detected by confocal spectral microscopy. The endometrial carcinoma cells （ HEC-1A, HEC-1A-PTEN-null, Ishikawa, Ishikawa-PTEN） were treated with signal transduetion inhibitors, RG-14620, SB203580 （SB） and rapamycin, respectively. Cell apoptosis morphology, cell apoptosis and cell cycle were detected by Hoechst 33258 staining and flow eytometry. Cell viability was determined by methyl thiazolyl tetrazolium assay. Results The PTEN protein expression in two endometrial carcinoma cells （Ishikawa, HEC-1A ） was exchanged by PTEN antisense oligonucleotide blocked and pcDNA3.1/PTEN stable transfected. After treated with RG-14620, SB and rapamycin, marked morphological changes of apoptosis were observed in HEC-1A-PTEN-null and Ishikawa cells. The cell apoptosis of HEC-1A-PTEN-null and Ishikawa cells exposed to SB were significantly increase [ （ 31.6 ± 0. 8 ） % and （ 37.8 ± 0. 8 ） %, respectively ], the G1 phase cells were increased to （ 84. 1 ± 3.2 ） % and （ 87.5 ± 1.9 ） %. While cell viability was significantly decreased in HEC-1A-PTEN-null and Ishikawa cells, the cell viability of HEC-1A-PTEN-null and Ishikawa cells exposed to SB were （54.0±2. 1 ） % and （49.0 ± 1.7 ） %. Conelusion Loss of PTEN in endometrial carcinoma cells may improve the G1 phase cells and apoptotic effects, inhibit the cell proliferation, which due to the sensitivity of cells to related signal transduction inhibitors.

关 键 词：子宫内膜肿瘤 PTEN磷酸水解酶 受体 表皮生长因子 西罗莫司 细胞凋亡 

分 类 号：R686[医药卫生—骨科学]