NS-398对人卵巢癌C13K细胞株增殖的抑制作用及对Snail和E-cadherin表达的调节

Effects of the selective inhibitor of COX-2 NS-398 in human ovarial carcinoma cell line C13K and its influence on the expression of Snail and E-cadherin

目的:探讨选择性环氧合酶-2(cyclooxygenase-2,COX-2)抑制剂NS-398对卵巢癌C13K细胞株的增殖抑制作用以及对Snail和E-cadherin基因表达的影响。方法:体外培养卵巢癌C13K细胞,不同浓度的NS-398(50μmol/L、100μmol/L、200μmol/L)作用于C13K细胞48h、72h后,MTT法分析不同浓度NS-398对细胞增殖的影响;RT-PCR法检测胞内Snail mRNA、E-cadherin mRNA的表达水平;免疫组化SABC法检测Snail、E-cadherin蛋白的表达。结果:NS-398对卵巢癌C13K细胞株增殖的抑制作用呈现时间和浓度依赖性,与对照组相比有统计学差异(P<0.05)。Snail mRNA和蛋白的表达随NS-398浓度的升高和作用时间的延长,逐渐降低;而E-cadherin mRNA和蛋白的表达则逐渐升高,与对照组相比有统计学差异(P<0.05)。结论:选择性COX-2抑制剂NS-398可明显抑制C13K细胞株的增殖,其机制可能与Snail和E-cadherin mRNA表达有关,为卵巢癌的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 in human ovarial carcinoma cell line C13K in vitro,and to explore its influence on the expression of snail and E-cadherin.Methods： Ovarial cancer cell C13K were cultured in vitro under the irritation of NS-398 with different concent rations（50,100,200μmol/L） for 48h and 72h.The inhibition rates were detected by methyl thiazolyl tetrazolium（MTT）.The expressions of Snail mRNA and E-cadherin mRNA were detected by Real-time PCR,the Snail protein and E-cadherin protein were detected by immunohistochemical method（SABC）.Results： The results of MTT showed that NS-398 significantly inhibited the proliferation of ovarial cancer cell in a dose and time dependentmanner.NS-398 at 200μmol/L obviously inhibited the proliferation of C13K cell,and there was significant difference between the experimental groups and the control groups（P〈0.05）.By RT-PCR and immunohistochemical method,the alteration of Snail mRNA and protein decreased after administration of NS398 for 48h and 72h,but the alteration of E-cadherin mRNA and protein increased.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 of overial cancer cell C13K via downregulating the expression of Snail and upregulating the expression of E-cadherin.