CAJ | 학술논문

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。
목적：탐토Chk1기인침묵대인란소암세포A2780방료민감성적영향。방법구건Chk1기인적단발협RNA（shRNA）질립전염인란소암세포A2780，동시이전염공재체화미전염조작위대조조，운용RT-PCR、단백면역인적방법관찰전염전후Chk1기인표체적차이；사갑기우담서람시험회제세포생장곡선；극륭형성시험관찰세포적방사민감성변화。결과여대조조상비，구건shRNA표체재체가억제인란소암세포A2780중Chk1 mRNA전록화단백적표체；Chk1기인침묵후세포생장명현만우대조조；병통과해제G2/M기조체，현저제고종류세포대방사선적민감성。결론파향Chk1적shRNA능유효억제인란소암세포A2780적Chk1기인표체，촉진세포조망，증강종류대방료적민감성。
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 .