chk2反义寡核苷酸对顺铂作用下HeLa细胞凋亡的影响

目的观察宫颈癌HeLa细胞转染chk2反义寡核苷酸后,在顺铂作用下对凋亡的影响。方法采用MTT法检测不同浓度顺铂作用人宫颈癌细胞株HeLa 24、487、2 h后对细胞的生长抑制率。以脂质体为载体将chk2正义链和反义链转染HeLa细胞,用Western blot检测HeLa细胞中Chk2蛋白的表达,用流式细胞仪和TUNEL法检测顺铂作用后细胞凋亡率。结果顺铂对HeLa细胞有生长抑制作用,并有时间和浓度依赖性;反义chk2转染后可明显抑制Chk2蛋白表达(P<0.01),顺铂作用下细胞凋亡率明显高于转染正义链组(P<0.01)。结论反义寡核苷酸封闭chk2基因可增加HeLa细胞对顺铂的凋亡敏感性。

Influence of antisense oligodeoxynucleotide targeting chk2 on apoptosis of HeLa cells induced by DDP

Objective To investigate the change of antisense oligodeoxynucleotide targeting chk2 apoptosis in cervical cancer cells HeLa induced by cisplastin(DDP).Methods The proliferation-inhibiting rates of HeLa cells induced by DDP in different concentration were measured by MTT assays.Oligonucleotide was transfected into HeLa cells by liposome.The expression of Chk2 was observed by Western blot.The apoptosis rate of HeLa cells induced by DDP was investigated by flow cytometer and TUNEL.Results The growth inhibition rate of HeLa cells increased gradually,which was dependent on the dosage of DDP and the action time.Transfection with antisense oligonuleotide targeting chk2 could inhibit the expression of Chk2 protein(P<0.01),and the apoptosis rate was significantly higher than that of the sense blocking(P<0.01).Conclusion Antisense blocking of chk2 could increase the apoptotic sensitivity of HeLa cells to DDP.