目的通过RNA干扰技术建立染色质重构蛋白CHD6基因表达抑制的细胞模型,研究CHD6对人肺腺癌细胞A549细胞增殖及辐射敏感性的影响.方法利用质粒介导的siRNA技术建立CHD6基因表达抑制细胞模型,RT-PCR检测CHD6 mRNA的表达,细胞生长曲线和流式细胞技术分别检测A549细胞增殖及细胞周期的变化,荧光染色法检测细胞凋亡,细胞克隆形成率检测A549细胞辐射敏感性.结果通过本研究,成功构建了siRNA抑制CHD6表达的细胞模型;通过siRNA抑制CHD6的表达,A549细胞增殖能力明显增强,细胞对2 Gy以内γ射线照射有明显辐射抗性;大剂量照射后的细胞凋亡率无明显改变.结论抑制CHD6基因表达将提高细胞增殖能力和细胞的辐射抗性.
Objective To observe the effects of siRNA-silenced chromatin reconstitution protein CHD6 on the proliferation and radiosensitivity in A549 cells. Methods CHD6-silenced cellular model was constructed by plasmid mediated siRNA technique. RT-PCR was used to detect the expression of mRNA. Growth curve, flow cytometry and fluorescent staining method were used to measure changes of cell proliferation, cell cycle and apoptosis, respectively. The radiosensitivity of A549 cells was determined with cell cloning efficiency test. Results siRNA-asilence CHD6 increased the growth of A549 cells and enhanced the radiation resistance of A549 cells when gamma irradiation dose was lesser than two grays, whereas it had no significant effect on apoptosis in cells being irradiated with larger doses. Conclusion Inhibition of CHD6 could enhance cell proliferation and cell radiosensitivitv.