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目的观察siRNA沉默NPM基因的表达对人皮肤鳞状细胞癌A431细胞增殖活性及细胞周期的影响。方法培养A431细胞,针对NPM mRNA序列设计合成特异性siRNA,转染A431细胞;采用实时定量PCR(Real-timePCR)法检测NPM基因的表达情况,CCK8法检测细胞的增殖活性,流式细胞术检测细胞周期变化。结果实时定量PCR法的结果显示:细胞NPM基因的表达受到明显抑制,并筛选出沉默效率最高的特异性siRNA片段进行后继实验;CCK8法检测结果显示NPM基因沉默后,细胞增殖活性显著降低(P<0.05);流式细胞术检测结果显示细胞发生S期阻滞,G2/M期细胞减少。结论沉默NPM基因的表达可明显抑制A431细胞的增殖活性,并导致细胞周期阻滞。提示NPM基因可能成为皮肤肿瘤防治的调控靶点。
Objective To observe the effects of siRNA silencing NPM gene on the proliferation and cell cycle of human skin squamous cell carcinoma A431 cells. Methods The A431 cells were cultured, and specific siRNAs were designed and synthesized based on the NPM mRNA sequence. Transfection of A431 cells was performed by Real-time PCR. Real-time PCR was used to detect the expression of NPM. CCK8 was used to detect the proliferation of cells. Flow cytometry Cell cycle changes. Results The results of real-time quantitative PCR showed that the expression of NPM gene was significantly inhibited and the specific siRNA fragment with the highest silencing efficiency was screened for subsequent experiments. The CCK8 assay showed that the NPM gene silencing significantly decreased the cell proliferation activity (P <0.05). Flow cytometry showed that S phase arrest occurred in cells and G2 / M phase cells decreased. Conclusion Silencing NPM gene expression can significantly inhibit the proliferation of A431 cells and lead to cell cycle arrest. Tip NPM gene may be the target of skin cancer prevention and control.