沉默EZH2基因表达对宫颈癌细胞增殖能力影响的分析

发布时间：2018-11-08 13:58

【摘要】：本实验旨在探讨EZH2基因在宫颈癌中表达情况、对宫颈癌细胞增殖能力的影响及其机制。通过免疫组织化学法检测20例正常宫颈、20例CIN及60例宫颈鳞癌组织中EZH2蛋白表达;采用RT-PCR法检测4个宫颈癌细胞株中EZH2 m RNA的表达。通过Lipofectamin2000介导EZH2 si RNA瞬时转染C33A细胞株,Western blotting法检测si RNA的干扰效率。MTT、流式细胞仪检测EZH2沉默后细胞增殖及细胞周期变化;Western blotting检测p21表达变化。结果显示,正常宫颈组织、CIN、宫颈鳞癌中EZH2阳性表达率分别为15%、30%、76.7%,呈逐级升高趋势,后者与前两者见差异有统计学意义(p0.01);EZH2的表达同宫颈鳞癌细胞分化程度、间质浸润深度及淋巴结转移显著相关(p0.05)。EZH2 m RNA在宫颈癌细胞株He La、Si Ha、C33A及Caski中均表达,其中C33A细胞中表达最高。si RNA沉默EZH2基因明显抑制宫颈癌C33A细胞增殖,阻滞细胞周期于G1期;Western blotting检测p21表达上调。由此得出结论,EZH2基因在宫颈癌中高表达;沉默EZH2基因可通过上调p21蛋白的表达阻滞细胞周期于G1期、抑制宫颈癌细胞的增殖。
[Abstract]:The aim of this study was to investigate the expression of EZH2 gene in cervical cancer and its mechanism. The expression of EZH2 protein in 20 cases of normal cervix, 20 cases of CIN and 60 cases of cervical squamous cell carcinoma were detected by immunohistochemical method, and the expression of EZH2 m RNA in 4 cervical cancer cell lines was detected by RT-PCR method. The interference efficiency of si RNA was detected by Lipofectamin2000 mediated EZH2 si RNA transient transfection of C33A cell line, Western blotting, and the expression of p21 was detected by MTT, flow cytometry after EZH2 silencing and; Western blotting expression. The results showed that the positive expression rate of EZH2 in normal cervical tissues and CIN, cervical squamous cell carcinoma was 1530 and 76.7respectively, which showed a gradual increase trend, and the difference between the latter and the former was statistically significant (p0.01). The expression of EZH2 was significantly correlated with the differentiation of cervical squamous cell carcinoma, the depth of interstitial infiltration and lymph node metastasis (p0.05). EZH2 m RNA was expressed in both He La,Si Ha,C33A and Caski cells. The highest expression of. Si RNA silencing EZH2 gene in C33A cells significantly inhibited the proliferation of cervical cancer C33A cells and blocked the cell cycle in G1 phase. The expression of p21 was up-regulated by Western blotting. It is concluded that EZH2 gene is highly expressed in cervical cancer and silencing EZH2 gene can inhibit the proliferation of cervical cancer cells by up-regulating the expression of p21 protein to block cell cycle in G1 phase.
【作者单位】：西安交通大学第一附属医院妇产科;
【基金】：陕西省国际科技合作与交流计划项目基金(2013KW-27-01) 中央高校基本科研业务费专项资金(XJJ2015-094)共同资助
【分类号】：R737.33

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