选择性环氧化酶-2抑制剂NS398诱导前列腺癌PC-3细胞系凋亡的研究

NS398 induces apoptosis of COX-2 negatively expressed human prostate cancer cell line PC-3 in vitro

目的 观察选择性环氧化酶2 (COX 2 )抑制剂氮2,环己氧4,硝基苯甲基磺胺(NS398)在诱导COX 2表达阴性的前列腺癌PC 3细胞凋亡中的作用。 方法 应用RT PCR和Westernblot的方法对PC 3细胞mRNA和蛋白水平的COX 2表达情况进行检测,四甲基偶氮唑蓝快速比色法观察不同浓度和时间NS398对PC 3细胞生存率的影响,流式细胞术检测100μmol/LNS398作用24hPC 3细胞的凋亡情况。 结果 mRNA和蛋白水平PC 3细胞COX 2均呈阴性表达;NS398可以抑制PC 3细胞的存活,并随浓度增加而增强,但无显著的时间依赖性;与对照组(10. 563±2. 582)%相比, 100μmol/LNS398诱导PC 3细胞早期凋亡比率(19. 307±3. 773)%显著增加,差异有统计学意义(P=0. 01)。 结论 选择性COX 2抑制剂NS398可以诱导COX 2表达阴性的PC 3细胞凋亡,提示COX 2非依赖性途径的存在。

Objective To determine whether the cyclooxygenase-2 (COX-2) selective inhibitor,{N-[2-(cyclohexyloxy)-4-nitrophenyl]-methane sulfonamide} (NS398),can inhibit proliferation of COX-2 negatively expressed human prostate cancer cell line PC-3 and induce its apoptosis in vitro. Methods The expression of COX-2 in cultured PC-3 cells was examined by RT-PCR and Western blot for mRNA and protein respectively.The proliferative inhibition of PC-3 cells by NS398 was observed by methyl thiazolyl tetrazolium (MTT) rapid photocolorimetric assay.PC-3 cells cultured in 100 μmol/L NS398 medium for 24 h was marked with AnnexinV-FITC and PI for the cell apoptosis assay by flow cytometry. Results COX-2 was not expressed in PC-3 cell line at either mRNA or protein level.The inhibitory rate of NS398 on proliferation increased with the increasing concentration but was not time-dependent.Compared with the controls (10.563±2.582)%,PC-3 cells cultured in 100μmol/L NS398 medium for 24 h had significantly higher incidence (19.307±3.773)%] <Abstract>Objective To determine whether the cyclooxygenase-2 (COX-2) selective inhibitor,{N-[2-(cyclohexyloxy)-4-nitrophenyl]-methane sulfonamide} (NS398),can inhibit proliferation of COX-2 negatively expressed human prostate cancer cell line PC-3 and induce its apoptosis in vitro. Methods The expression of COX-2 in cultured PC-3 cells was examined by RT-PCR and Western blot for mRNA and protein respectively.The proliferative inhibition of PC-3 cells by NS398 was observed by methyl thiazolyl tetrazolium (MTT) rapid photocolorimetric assay.PC-3 cells cultured in 100 μmol/L NS398 medium for 24 h was marked with AnnexinV-FITC and PI for the cell apoptosis assay by flow cytometry. Results COX-2 was not expressed in PC-3 cell line at either mRNA or protein level.The inhibitory rate of NS398 on proliferation increased with the increasing concentration but was not time-dependent.Compared with the controls (10.563±2.582)%,PC-3 cells cultured in 100μmol/L NS398 medium for 24 h had significantly higher incidence (19.307±3.773)%] of early apoptosis (P=0.01).Conclusions NS398 can inhibit proliferation of COX-2 negatively expressed human prostate cancer cell line PC-3 and induce its apoptosis in vitro,which suggests its COX-2 independent pathway.