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[目的]通过小干扰RNA(siRNA)沉默EGFR基因的表达,探讨其对卵巢癌SKOV3细胞体外侵袭力的影响。[方法]体外构建靶向EGFR基因的siRNA质粒和阴性对照质粒,Lipofectamine2000介导转染到SKOV3细胞中。实验分3组:空白对照组、非特异性转染组、特异性转染组。RT-PCR和免疫细胞化学法检测EGFR基因和蛋白水平表达。平板克隆形成法检测细胞生长增殖能力,Transwell侵袭实验检测细胞体外侵袭力。[结果]特异性转染组细胞EGFR基因mRNA和蛋白表达的抑制率分别为52.3%和51.8%,克隆形成抑制率和侵袭抑制率分别为47.4%和40.1%。[结论]siRNA可以有效抑制SKOV3细胞EGFR基因的表达,抑制其增殖能力和体外侵袭能力。
[Objective] To investigate the effect of EGFR gene silencing on the invasiveness of ovarian cancer SKOV3 cells by siRNA. [Method] siRNA plasmid and negative control plasmid targeting EGFR gene were constructed in vitro. Lipofectamine 2000 was transfected into SKOV3 cells. The experiment was divided into 3 groups: blank control group, non-specific transfection group, specific transfection group. The expression of EGFR gene and protein was detected by RT-PCR and immunocytochemistry. Cell cloning formation assay was used to detect cell proliferation and proliferation. Transwell invasion assay was used to detect cell invasiveness in vitro. [Results] The inhibitory rates of EGFR gene mRNA and protein expression in specific transfected cells were 52.3% and 51.8%, respectively. The rates of clonogenic and invasion inhibition were 47.4% and 40.1%, respectively. [Conclusion] siRNA can effectively inhibit the expression of EGFR gene in SKOV3 cells and inhibit its proliferation and invasion ability in vitro.