COX-2反义寡核苷酸抑制宫颈癌Hela细胞的研究

目的：通过研究环氧合酶-2（COX-2）反义寡核苷酸（AS-ODNs）对宫颈癌Hela细胞的增殖抑制及对其抑制原理进行分析,探讨COX-2 AS-ODNs对宫颈癌的治疗意义.方法：Hela细胞分6组培养：对照组（A）、正义寡核苷酸（SODNS）组（B）、脂质体组（C）及反义寡核苷酸组 （D）设3个亚组.MTT实验检测细胞的增殖情况;在转染后48 h,RT-PCR法检测COX-2 mRNA的表达水平,Western blot检测COX-2蛋白的表达,流式细胞仪测定细胞周期的变化.结果：与其他组比较,AS-ODNs转染的细胞增殖缓慢,且随着AS-ODNs的浓度增加细胞增殖指数有下降的趋势,COX-2 mRNA及蛋白表达明显下调,表达量随AS-ODNs转染浓度的增加而减少,细胞周期S期细胞比例明显减少,G2/M及G0/G1期细胞所占比例增多,P ＜0.05.结论：COX-2反义寡核苷酸对Hela细胞的增殖有抑制作用;其抑制机制可能与抑制COX-2转录与翻译及细胞周期再分布有关.

Study of inhibition of Hela cell of cervical carcinoma by cyclooxygenase-2 antisense oligodeoxynucleotides

OBJECTIVE:To investigate the effect of the antisense oligodeoxynucleotides(AS-ODNs) of human cyclooxygenase-2 (COX-2) inhibit cervical cancer Hela cell line proliferation, and explore its mechanism and discuss its significance of treat cervical cancer. METHODS: The Hela cells were cultured and divided into 6 groups: Control group(A), Sense oligodeoxynucleotides(SODNS) group(B), Lipofectin group(C), AS-ODNs group including three sub-groups.The proliferative status of cells was measured by methabenzthiazuron (MTT) assay to detect the alive Hela cells; in the Hela cells,RT-PCR was used to observe the expression of COX-2 mRNA;in the way of Western blotting, the expression of COX-2 protein was observed; cell cycle were analyzed by flow cytometry.RESULTS: Compared with other groups, the proliferation of Hela cells was inhibited in the COX-2 ASODN group,the proliferative index decreased significantly with the increase of COX-2 AS-ODNs concentration. RT-PCR showed that the down regulation of expression of COX-2 mRNA with the increase of the COX-2 AS-ODN concentration; Western blot showed that the expression COX-2 protein was inhibited,in the cell cycle, the percentage of S phase decreased remarkably (P<0.05) and the percentage of G2/M and G0/G1 phase increased (P<0.05) in AS-ODNs transfected cells compared with that in the control group cells. CONCLUSION:Transfection with antisense AS-ODNs of human COX-2 mRNA could inhibit the proliferation of human cervical cancer Hela cells, and its inhitit michenism probably is inhibiting COX-2 expression in mRNA and protein level and to lead to cell cycle redistributing.