NS-398对宫颈癌细胞系的增殖抑制作用及对survivin基因表达的影响

Effects of the selective inhibitor of COX-2 NS-398 in cervical cancer cells and its influence on the expression of survivin mRNA

目的:研究选择性环氧合酶-2(COX-2)抑制剂NS-398对宫颈癌HeLa,SiHa细胞系的增殖、凋亡作用及其对凋亡抑制基因survivin表达的影响。方法:体外培养宫颈癌HeLa,SiHa细胞系,用四甲基偶氮唑蓝(MTT)比色法分析不同浓度的NS-398分别作用于HeLa,SiHa细胞系24h、48h后对细胞增殖的作用;流式细胞仪(FCM)检测对细胞凋亡的作用;RT-PCR分析对凋亡抑制基因survivin表达的影响。结果:MTT检测显示,NS-398可抑制宫颈癌HeLa,SiHa细胞系增殖,并有浓度时间依赖性,与对照组相比差异有统计学意义(P<0.05)。FCM检测提示,NS-398可诱导HeLa,SiHa细胞系凋亡,有浓度依赖性,与对照组相比差异有统计学意义(P<0.05)。RT-PCR分析表明,NS-398可抑制HeLa,SiHa细胞系凋亡抑制基因survivinmRNA表达,与对照组的差异有统计学意义(P<0.05)。结论:NS-398可抑制宫颈癌HeLa,SiHa细胞增殖,诱导凋亡,其机制与抑制凋亡抑制基因survivin mRNA表达有关,为宫颈癌治疗提供了新的靶点和理论依据。

Objective： To investigate the effects of the selective inhibitor of cyclooxygenase-2（COX-2） NS-398 on the proliferation and the pro-apoptosis in human cervical carcinoma cell lines HeLa and SiHa in vitro, and to explore its influence on the expression of survivin mRNA. Methods：Cervical cancer cells HeLa and SiHa were cultured in vitro respectively under the irritation of NS-398 with different concent rations （25,50, 100μmol/L） for 24h and 48h. The inhibition rates were detected by methyl thiazolyl tetrazolium （MTF）. Flow cytometry （FCM） was prepared to determine its pro-apoptosis activity at 48h after the administration of NS-398 with different concentrations （25,50,100txmol/L）. The expressions of survivin mRNA were detected by reverse transcription-polymerase chain reaction（RT-PCR） at 48h after admin istrated with NS-398 of different concentrations （25,50,100μmol/L）. Results： The results of MTF showed that NS-398 significantly inhibited the proliferation of cervical cancer cells in a dose and time dependent manner. NS-398 at 100μmol/L obviously inhibited the proliferation of HeLa and SiHa cells, and there was significant difference between the experimental groups and the control groups （P 〈 0. 05 ）. By FCM, NS-398 could induce apoptosis, especially at 100μmol/L. The difference was significant between the experimental groups and the control groups （ P 〈 0.05 ）. Using RT-PCR, the alteration of survivin mRNA decreased after administration of NS-398 for 48h. The difference was significant between the experimental groups and the control groups. （ P 〈 0.05 ）. Conclusion： The selective inhibitor of COX-2 NS-398 can inhibit proliferation and induce the development of apoptosis of cervical cancer cells HeLa and SiHa via downregulating the expression of survivin mRNA.