STOML-2过表达抑制人宫颈鳞癌Siha细胞凋亡  

Overexpression of STOML-2 inhibits apoptosis of human cervical squamous carcinoma Siha cells in vitro

作　　者：胡国林[1] 姚广裕[1] 邓欢[2] 赵可雷[3] 许斐斐[1] 梁卫江[1] 刘国炳[1] 

机构地区：[1]南方医科大学南方医院,广东广州510515 [2]南方医科大学第三附属医院,广东广州510631 [3]新乡医学院第一附属医院,河南新乡453199

出　　处：《南方医科大学学报》2015年第9期1293-1296,共4页Journal of Southern Medical University

基　　金：广东省自然科学基金(S2013010016194)

摘　　要：目的研究STOML-2过表达抑制人宫颈鳞癌Siha细胞凋亡的机制,以期为宫颈癌的治疗开辟新的路径。方法用携带STOML-2基因的腺病毒感染Siha细胞作为过表达组,同时设立空白载体组及空白组作为对照,72 h后用Western blot检测STOML-2的过表达,并用MTT法检测STOML-2过表达对细胞增殖的影响。用IC50的顺铂处理各组细胞24 h后,利用荧光显微镜观察STOML-2过表达对细胞凋亡的影响,并利用流式细胞仪进一步分析细胞的凋亡率;采用Western blot检测线粒体凋亡途径相关蛋白caspase3、cleaved-caspase3、Bcl-2、Bax以及线粒体内外细胞色素C(Cyt C)表达量的变化。结果与空白组及空白载体组相比,Western blot结果显示过表达组STOML-2的表达量明显增加;MTT结果显示,STOML-2过表达后可明显促进Siha细胞的增殖。用IC50的顺铂处理细胞24 h后,荧光显微镜下,空白载体组凋亡细胞的细胞核大小不一,浓染致密的蓝色荧光显示核固缩,部分核裂解为凋亡小体,而过表达组细胞核多较完整,着色较浅,染色质密度均匀一致,并且流式细胞仪结果同样提示STOML-2过表达后细胞凋亡率明显降低。Western blot结果显示过表达组线粒体凋亡途径相关蛋白cleaved-caspase3、Bax以及线粒体外Cyt C表达水平降低,而caspase3、Bcl-2及线粒体内Cyt C的表达水平则相应升高。结论 STOML-2过表达可以促进Siha细胞增殖,其抑制凋亡的机制可能与线粒体凋亡途径相关。Objective To investigate the mechanism underlying the inhibitory effect of STOML-2 overexpression on apoptosis of human cervical squamous carcinoma Siha cells. Methods Siha cells were transfected with an adenoviral vector carrying STOML- 2, and 72 h later STOML- 2 expression and the proliferation of the cells were detected by Western blotting and MTT assay. The transfected cells were treated with IC50 Cisplatin for 24 h, and the morphological changes of cells were observed using fluorescence, and the cell apoptosis was analyzed using flow cytomerty; the expression levels of proteins related with mitochondrial apoptosis pathway, including caspase-3, cleaved caspase-3, Bcl-2, Bax and cytochrome C（Cyt C）, were detected by Western blotting. Results Western blotting showed a significantly increased STOML- 2 expression in the transfected cells.Overexpression of STOML-2 obviously promoted the proliferation of Siha cells. The STOML-2-overexpressing cells exhibited an obvious resistance to IC50Cisplatin-induced apoptosis as shown by both fluorescence microscopy and flow cytometry and presented with decreased expressions of cleaved caspase-3, Bax, and cytosol Cyt C and increased expressions of caspase-3, Bcl-2, and mitochondrial Cyt C. Conclusion Overexpression of STOML-2 can enhance the proliferation of Siha cells by inhibiting cell apoptosis possibly through the mitochondrial apoptosis pathway.

关 键 词：宫颈鳞癌 STOML-2 线粒体凋亡途径 

分 类 号：R737.33[医药卫生—肿瘤]