金属硫蛋白反义寡核苷酸对子宫颈癌细胞增殖及凋亡的影响

Effects of metallothionein antisense oligodeoxynucleotide on the biological behaviors of HeLa cervical cancer cell line

目的:探讨金属硫蛋白-2A(MT-2A)反义寡核苷酸(ASODN)对子宫颈癌HeLa细胞体外生物学行为的影响。方法:设空白对照组、正义寡核苷酸(SODN)组及ASODN组,应用RT-PCR和Western Blot检测MT-2A mRNA和蛋白表达水平的改变;四甲基偶氮唑蓝(MTT)法检测HeLa细胞的增殖抑制率;流式细胞技术检测HeLa细胞凋亡率及细胞周期。结果:MT-2A ASODN对MT-2A mRNA表达抑制率为70%,并可显著抑制MT-2A蛋白的表达;ASODN组HeLa细胞增殖的抑制率为56.45%,凋亡指数(AI)为(16.76±2.08)%,S期细胞百分率为(11.93±2.08)%,与空白对照组、SODN组比较,差异均有统计学意义(P<0.05)。结论:MT-2A ASODN可有效抑制子宫颈癌HeLa细胞中MT-2A mRNA和蛋白的表达,促进细胞凋亡并阻止细胞进入S期而抑制细胞增殖,MT-2A可能参与了子宫颈癌的发生发展。

Objective： To investigate effects of metallothionein-2A （MT-2A） antisense oligodeoxynucleotide（ASODN） on the biological behavior of a human cervical cancer cell line HeLa in vitro. Methods ：The expression of MT-2A was detected by RT-PCR and Western blot. The proliferation inhibitory rate of HeLa was assessed by MTF, and the apoptotic changes were examined by flow cytometry, respectively. Results：The expression inhibitory rate of MT-2A mRNA was 70%, the expression of MT-2A protein was decreased significantly. In the group of ASODN,the proliferation inhibitory rate of HeLa cells was 56.45%, and the apoptosis index and percentage of cells in S phase were （ 16.76 ±2.08 ） % and （ 11.93 ±2.08 ） %, respectively. There were significant differences as compared with the vacuity control group and the SODN group（ P 〈 0.05 ）. Conclusions ： MT-2A ASODN can effectively inhibit the expression of MT-2A mRNA and protein, enhance apoptosis and increase cell proliferation by arrest S phase entrance. MT-2A may participate in the occurrence and development of cervical cancer.