南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (06): 866-.

• • 上一篇    下一篇

siRNA靶向EZH2 抑制子宫内膜癌细胞生长和侵袭

冷灵芝,黄启涛,董玉楠,俞泓,田焱,彭国庆   

  • 出版日期:2013-06-20 发布日期:2013-06-20

EZH2 gene silenced by siRNA suppresses the growth and invasion of endometrial carcinoma cells

  • Online:2013-06-20 Published:2013-06-20

摘要: 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。

Abstract: Objective To explore the effects on cell proliferation and invasion as well as molecular basis after suppressing EZH2
expression in endometrial carcinoma cells by using siRNAs. Methods RT-PCR was used to examine the expression of EZH2 in
endometrial carcinoma and their paracancerous tissues. SiRNAs targeting to EZH2 were transfected to endometrial carcinoma
cells, and MTT, FACS, and boyden assays were utilized to examine cell proliferation, cell cycle change, and cell invasion.
Finally, the molecular mechanisms of EZH2 on cell function alteration were investigated. Results Compared with
paracancerous tissues, increased expression trend of EZH2 mRNA was showed in endometrial carcinoma tissues. Further,
knocking down EZH2 expression inhibited cell growth, cell cycle transition from G1 to S phase, and cell invasion ability.
Molecular basis indicated that suppression of EZH2 downregulated the expression of E2F1 and MMP9 and upregulated tumor
suppressor p21 expression. Conclusions EZH2 expression is increased in endometrial carcinoma tissues. Knocking down
EZH2 expression suppresses the cell growth, cell cycle transition and cell invasion by downregulated E2F1 and MMP9, and
upregulated tumor suppressor p21 expression.