RUNX2在宫颈鳞癌组织中的表达及其siRNA对Hela229细胞RUNX2表达及细胞增殖与凋亡的影响  

Expression of RUNX2 in human cervical squamous cell carcinoma tissue and effect of RUNX2 siRNA on expression of RUNX2,proliferation and apoptosis of Hela229 cells

作　　者：张威[1] 郭文涛 曾宪旭[1] 班振英[1] 

机构地区：[1]郑州大学第三附属医院病理科,郑州450052 [2]广东医科大学基础医学院病原生物实验室,广东东莞523808

出　　处：《郑州大学学报（医学版）》2017年第6期718-722,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省高等学校重点科研计划项目17A310027

摘　　要：目的:探讨RUNX2蛋白在宫颈鳞癌组织中的表达情况及RUNX2 siRNA对宫颈鳞癌Hela229细胞RUNX2基因表达、细胞增殖及凋亡的影响。方法:应用免疫组化技术检测50例宫颈鳞癌、30例宫颈高级别鳞状上皮内病变(HSIL)及30例正常宫颈黏膜组织中RUNX2蛋白的表达情况,并分析宫颈鳞癌组织中RUNX2蛋白表达与临床病理特征的关系。设计合成RUNX2 siRNA,通过脂质体将其转染至Hela229细胞,同时设立空白对照(不转染)和阴性对照(转染非特异性siRNA)细胞组。转染24、48、72 h后,采用MTT法检测各组细胞增殖,计算增殖抑制率;转染72 h后,采用RT-PCR及Western blot技术检测细胞中RUNX2 mRNA及蛋白的表达,采用流式细胞术检测细胞凋亡。结果:宫颈鳞癌组织中RUNX2阳性表达率高于HSIL及正常宫颈组织(P<0.001);宫颈鳞癌组织中RUNX2蛋白的表达与肿瘤的临床分期有关(P=0.004)。与空白对照组和阴性对照组比较,RUNX2 siRNA组Hela229细胞增殖抑制率、RUNX2 mRNA及蛋白的表达均降低(P<0.05),细胞凋亡率升高(P<0.05)。结论:RUNX2基因的高表达与宫颈鳞癌细胞的增殖关系密切。Aim：To explore the expression of RUNX2 protein in cervical squamous cell carcinoma tissue and the effect of RUNX2 siRNA on the expression of RUNX2,cell proliferation and apoptosis in Hela229 cells.Methods：Immunohistochemical technique was used to measure the expression of RUNX2 protein in 50 cases of cervical squamous cell carcinoma,30 cases of cervical high-grade squamous intraepithelial lesion（HSIL）and 30 normal cervical epitheliums（NCE）tissue,and the relationship between the RUNX2 protein expression and the clinical pathological parameters was analyzed.RUNX2siRNA was synthesized,and transfected into Hela229 cells by liposome method.MTT method was used to detect the cell proliferation after 24,48 and 72 h culture;the mRNA and protein expressions of RUNX2 were measured respectively by RT-PCR and Western blot,and flow cytometry was used to detect cell apoptosis after 72 h culture.Results：The RUNX2 positive expression rate in the cervical squamous cell carcinoma tissue was higher than those in HSIL or NCE tissue（P〈0.01）,and RUNX2 protein positive expression rate was associated with the clinical stage of tumor（P=0.004）.Compared with the two control groups,the proliferation inhibition rate of RUNX2 siRNA group,and the expressions of RUNX2 mRNA and protein at 72 h were lower（P〈0.05）,while the apoptosis index was higher（P〈0.05）.Conclusion：There is close correlation between high expression of RUNX2 gene and proliferation of cervical cancer cells.

关 键 词：宫颈癌 HELA 229细胞 RUNX2 RNA干扰 增殖 凋亡 

分 类 号：R737.3[医药卫生—肿瘤]