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目的探讨靶向抑制EZH2表达对子宫内膜癌细胞生长及侵袭的影响及其分子机制。方法实时荧光定量PCR检测EZH2在子宫内膜癌与癌旁子宫内膜组织中的表达差异。利用化学合成siRNA转染子宫内膜细胞,靶向抑制EZH2表达。MTT、流式细胞仪检测细胞增殖和细胞周期改变。Boyden小室检测细胞侵袭情况的变化。Western blotting检测细胞周期因子及基质金属蛋白酶2(MMP2)表达改变。结果相比于癌旁子宫内膜组织,EZH2基因在子宫内膜癌中表达相对增高。SiRNA在子宫内膜癌细胞中抑制EZH2表达后,细胞增殖能力明显降低,且细胞周期也阻滞在G1期。Boyden小室分析显示,在抑制EZH2表达后,细胞的侵袭能力也明显降低。机制分析显示,在抑制EZH2表达后,细胞周期因子E2F1及MMP2表达明显降低,而抑癌基因p21表达明显升高。结论 EZH2在子宫内膜癌中表达明显升高。SiRNA靶向抑制EZH2之后通过下调E2F1、MMP2及上调p21表达后,能明显减慢子宫内膜癌细胞增殖和侵袭速度。
Objective To investigate the effects of targeted inhibition of EZH2 expression on the growth and invasion of endometrial carcinoma cells and its molecular mechanism. Methods The expression of EZH2 in endometrial carcinoma and adjacent normal endometrium was detected by real-time fluorescence quantitative PCR. Transfection of endometrial cells with chemically synthesized siRNAs targeted the inhibition of EZH2 expression. MTT, flow cytometry to detect cell proliferation and cell cycle changes. Boyden chamber to detect changes in cell invasion. Western blotting was used to detect the expression of cell cycle factor and matrix metalloproteinase 2 (MMP2). Results Compared with the adjacent endometrial tissue, EZH2 gene expression in endometrial cancer increased relatively. SiRNA inhibition of EZH2 expression in endometrial cancer cells, cell proliferation decreased significantly, and the cell cycle is also arrested in the G1 phase. Boyden chamber analysis showed that after inhibiting EZH2 expression, cell invasion ability was also significantly reduced. Mechanism analysis showed that after inhibiting EZH2 expression, the expression of cell cycle factor E2F1 and MMP2 was significantly decreased, while the expression of tumor suppressor gene p21 was significantly increased. Conclusion EZH2 expression in endometrial cancer was significantly higher. SiRNA targeted inhibition of EZH2 can significantly slow down the proliferation and invasion of endometrial carcinoma cells by down-regulating E2F1, MMP2 and up-regulating p21 expression.