chk1反义寡核苷酸增加HeLa细胞顺铂作用下凋亡敏感性

Antisense oligodeoxynucleotide targeting chk1 increases sensitivity of HeLa cells to apoptosis induced by DDP

目的:通过反义封闭chk1基因,阻断细胞周期检测点信号传导通路,从而增加宫颈癌HeLa细胞对顺铂(DDP)的敏感性.方法:采用MTT法检测梯度浓度DDP作用人宫颈癌细胞株HeLa24,48和72h后对细胞的生长抑制率.以脂质体为载体将chk1正义链和反义链转染HeLa细胞,用Western Blot测量Chk1蛋白的表达,用流式细胞仪和TUNEL法检测DDP作用后细胞凋亡率.结果:DDP对HeLa细胞有生长抑制作用,并有时间和浓度依赖性.反义封闭chk1基因可抑制Chk1蛋白表达(P<0.01),DDP作用下细胞凋亡率为54.1%,高于转染正义链的34.2%(P<0.01).结论:反义封闭chk1基因可增加HeLa细胞对DDP的凋亡敏感性.

AIM： To investigate the effect of chkl gene in cisplatin-resistance of cervical cancer to block the signal transduction pathways of cell cycle checkpoint, by targeting chkl gene using antisense oligonucleotide, so as to enhance the drug sensitivity of cervical cancer HeLa cells to DDP. METHODS： The proliferation inhibition rates of HeLa cells by DDP in different concentrations were measured by MTT assay. Oligonucleotide was transfected into HeLa cells. The expression of Chkl was observed by Western Blot. The apoptosis rate of HeLa cells induced by DDP was investigated by flow cytometry and TUNEL. RESULTS： The growth inhibition rate of HeLa cells increased gradually in a time and concentration dependent manner. Transfection with antisense oligonuleotide targeting chkl inhibited the expression of Chkl protein （P 〈0.01 ）, and the apoptosis rate （54.13%） was significantly higher than that of the sense blocking （34.20% , P 〈 0.01 ）. CONCLUSION： Antisense blocking of chkl increases the apoptosis sensitivity of HeLa cells to DDP.