Chk1基因沉默对人卵巢癌细胞 A2780放疗敏感性的影响

摘要

目的：探讨Chk1基因沉默对人卵巢癌细胞A2780放疗敏感性的影响。方法构建Chk1基因的短发夹RNA（shRNA）质粒转染人卵巢癌细胞A2780，同时以转染空载体和未转染组作为对照组，运用RT-PCR、蛋白免疫印迹方法观察转染前后Chk1基因表达的差异；四甲基偶氮唑蓝试验绘制细胞生长曲线；克隆形成试验观察细胞的放射敏感性变化。结果与对照组相比，构建shRNA表达载体可抑制人卵巢癌细胞A2780中Chk1 mRNA转录和蛋白的表达；Chk1基因沉默后细胞生长明显慢于对照组；并通过解除G2/M期阻滞，显著提高肿瘤细胞对放射线的敏感性。结论靶向Chk1的shRNA能有效抑制人卵巢癌细胞A2780的Chk1基因表达，促进细胞凋亡，增强肿瘤对放疗的敏感性。%Objective To investigate the effect of Chk 1 gene silencing on the proliferation and radiosensitivity of hu-man ovarian cell line A2780.Methods Vectors containing short hairpin RNA (shRNA) targeting Chk1 gene were con-structed and transfected into human ovarian carcinoma cell line A 2780 .At the same time , transfecting the empty vector and untransfected group were selected as controls .The RT-PCR and Western blotting were used to compare the differences of Chk1 expression before and after trasfection .The cell growth curve was drawn by methyl thiazolyl tetrazolium ( MTT);and the radiosensitivity of ovarian carcinoma was evaluated by Clone formation array .Results Compared with the control group, constructing shRNA expression vector inhibited the Chk 1 mRNA transcription and protein expression of human ovari-an carcinoma cell line A2780;after Chk1 gene silencing , the cell growth rate was obviously slower than that of the control group;the radiosensitivity of A2780 cells was enhanced by decreasing the G 2/M phase blocking.Conclusion The shR-NA targeting at Chk1 gene can efficiently inhibit the Chk 1 expression in A2780 cells, which can enhance the apoptosis and obviously increase the radiosensitivity in human ovarian cell lines .