论文部分内容阅读
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(p<0.01);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(p<0.05)。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。
The purpose of this study is to investigate the expression of EZH2 in cervical cancer and its effect on the proliferation of cervical cancer cells and its mechanism. The expression of EZH2 protein was detected by immunohistochemistry in 20 cases of normal cervical tissue, 20 cases of CIN and 60 cases of cervical squamous cell carcinoma. The expression of EZH2 mRNA in 4 cervical cancer cell lines was detected by RT-PCR. C33A cells were transiently transfected with EZH2 si RNA by Lipofectamin2000, and the interference efficiency of si RNA was detected by Western blotting. MTT and flow cytometry were used to detect cell proliferation and cell cycle changes after EZH2 silencing. The expression of p21 was detected by Western blotting. The results showed that the positive expression rates of EZH2 in normal cervical tissue, CIN and cervical squamous cell carcinoma were 15%, 30% and 76.7%, respectively, showing a step-by-step upward trend. The difference between the latter and the former two was statistically significant (p < 0.01). The expression of EZH2 was significantly correlated with the differentiation of cervical squamous cell carcinoma, the depth of interstitial infiltration and lymph node metastasis (p <0.05). EZH2 m RNA was expressed in cervical cancer cell lines He La, Si Ha, C33A and Caski, of which C33A cells were the highest. The silencing of EZH2 gene by si RNA significantly inhibited the proliferation of C33A cells and blocked the cell cycle in G1 phase. The expression of p21 was up-regulated by Western blotting. It was concluded that EZH2 gene was highly expressed in cervical cancer. Silencing EZH2 gene could block the cell cycle in G1 phase by up-regulating the expression of p21 protein and inhibit the proliferation of cervical cancer cells.