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

发布时间：2018-06-27 03:06

本文选题：宫颈癌 + Hela　；参考：《郑州大学学报(医学版)》2017年06期

【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(P0.001);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(P0.05),细胞凋亡率升高(P0.05)。结论:RUNX2基因的高表达与宫颈鳞癌细胞的增殖关系密切。
[Abstract]:Objective: to investigate the expression of RUNX2 protein in cervical squamous cell carcinoma (SCC) and the effect of RUNX2 siRNA on RUNX2 gene expression, cell proliferation and apoptosis in Hela229 cells. Methods: the expression of RUNX2 protein in 30 cases of high grade squamous intraepithelial lesions (HSIL) of cervix and 30 cases of normal cervical mucosa were detected by immunohistochemical technique. The relationship between the expression of RUNX2 protein and clinicopathological features of cervical squamous cell carcinoma was analyzed. RUNX2 siRNAs were designed and synthesized and transfected into Hela229 cells by liposome. After transfection for 72 h, the proliferation of the cells was detected by MTT assay, and the expression of RUNX2 mRNA and protein was detected by RT-PCR and Western blot, and apoptosis was detected by flow cytometry. Results: the positive expression rate of RUNX2 in cervical squamous cell carcinoma was higher than that in HSIL and normal cervix (P0.001), and the expression of RUNX2 protein was related to the clinical stage of cervical squamous cell carcinoma (P0. 004). Compared with the control group and the negative control group, the inhibitory rate of proliferation and the expression of RUNX2 mRNA and protein of Hela229 cells in RUNX2 siRNA group were decreased (P0.05), and the apoptosis rate was increased (P0.05). Conclusion the high expression of RUNX2 gene is closely related to the proliferation of cervical squamous carcinoma cells.
【作者单位】：郑州大学第三附属医院病理科;广东医科大学基础医学院病原生物实验室;
【基金】：河南省高等学校重点科研计划项目17A310027
【分类号】：R737.33

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