一种选择性环氧化酶2—抑制剂JTE—522抑制RL95—2细胞的增殖及诱导其凋亡

目的:探讨环氧化酶-2抑制剂JTE-522是否对人子宫内膜癌细胞株RL95-2细胞有抑制增殖和诱导凋亡的作用及其分子机理.方法:应用体外噻唑兰法、琼脂糖凝胶电泳、酶联免疫试验、流式细胞术、RT-PCR及Western blot等方法研究JTE-522对RL952细胞增殖和凋亡的作用及其分子机理.结果:JTE522抑制RL95-2细胞的增殖、诱导其凋亡、引起G_0/G_1期阻滞和 S期抑制,并伴有COX-2 mRNA、磷酸化 Rb、CDK4蛋白表达的抑制及 p53,p21和cyclin D_1蛋白表达水平的上调.另外,细胞经 JTE-522处理后,还可见caspase-3活性的增加.结论:JTE-522抑制RL95-2细胞的增殖及诱导其凋亡,可能与COX-2mRNA,磷酸化Rb、CDK4蛋白水平的下降及 p53,p21和 cyclin D_1蛋白表达的上调有关,还可能与caspase-3的激活有关.

JTE-522, a selective COX-2 inhibitor,inhibits cell proliferation and induces apoptosis in RL95-2 cells

AIM: To investigate whether JTE-522 [4-(4-cyclohexyl-2-methyloxazol-5-yl )-2-fluorobenzenesulfonamide], a selective COX-2 inhibitor, can induce apoptosis and inhibit cell proliferation in human endometrial cancer cell line RL95-2 cells and to explore the molecular mechanisms. METHODS: [3-(4,5)-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT), DNA ladder, enzyme-linked immunosorbent assay (ELISA), flow cytometry, RT-PCR, and Western blot analysis were employed to investigate effect of JTE-522 on human endometrial cancer cell line RL95-2 cells and the related molecular mechanisms. RESULTS: JTE-522 inhibited the growth of RL95-2 cells and induced the apoptosis. Furthermore, it arrested G0/G1 phase and inhibited S phase in RL95-2 cells. JTE-522 inhibited the expressions of COX-2 mRNA, phosphorylated Rb, and CDK4 proteins, while increased the levels of p53, p21, cyclin D1 proteins, and the activity of caspase-3 in RL95-2 cells. CONCLUSION: JTE-522 inhibits cell proliferation and induces apoptosis in RL95-2 cells, which may be associated with the activation of caspase-3-like proteases, down-regulation of the expression of COX-2 mRNA, phosphorylated Rb, and CDK4 proteins, and up-regulation of the expressions of p53, p21, and cyclin D1 proteins.