摘要
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-2AS-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48h,RT-PCR法检测COX-2mRNA的表达水平,Westernblot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-2mRNA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。
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 ceils 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 Go/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.
出处
《中华肿瘤防治杂志》
CAS
2007年第4期273-276,共4页
Chinese Journal of Cancer Prevention and Treatment
