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目的研究NS-398(一种选择性环氧合酶-2抑制剂)对结肠癌HT-29细胞的生长抑制作用并探讨其机制。方法通过MTT法检测细胞的增殖情况,通过流式细胞术检测细胞凋亡率和细胞周期,通过RT-PCR检测bcl-2mRNA和baxmRNA表达,通过共聚焦激光扫描显微镜观察细胞骨架成分F-actin的变化。结果NS-398 对结肠癌HT-29细胞生长的抑制具有时间和剂量依赖性。流式细胞术结果显示NS-398可剂量依赖地诱导HT- 29细胞凋亡,使其停滞于G0/G1期。经不同浓度的NS-398处理72h后,bcl-2mRNA在HT-29细胞的表达降低, bcl-2/bax比值降低。细胞骨架成分F-actin主要分布在HT-29细胞核周围,呈环状结构,NS-398作用后细胞核周围的环状结构消失。结论NS-398可显著抑制结肠癌细胞的体外生长并诱导其凋亡,这与下调bcl-2/bax比值以及细胞骨架的破坏有关。
Objective To investigate the inhibitory effect of NS-398 (a selective cyclooxygenase-2 inhibitor) on the growth of colon cancer HT-29 cells and to explore its mechanism. Methods The cell proliferation was detected by MTT assay. The apoptosis rate and cell cycle were detected by flow cytometry. The expressions of bcl-2 mRNA and bax mRNA were detected by RT-PCR. The expression of F-actin in cytoskeleton was observed by confocal laser scanning microscope Variety. Results The inhibitory effect of NS-398 on the growth of HT-29 colon cancer cells was time-and dose-dependent. Flow cytometry results showed that NS-398 induced apoptosis in HT-29 cells in a dose-dependent manner and arrested at G0 / G1 phase. The expression of bcl-2mRNA in HT-29 cells decreased and the ratio of bcl-2 / bax decreased after 72h treatment with different concentrations of NS-398. Cytoskeletal components F-actin mainly distributed in the nucleus of HT-29 cells, showing a ring-shaped structure, and the ring-shaped structure disappeared around the nucleus after the action of NS-398. Conclusion NS-398 can significantly inhibit the growth of colon cancer cells in vitro and induce its apoptosis, which is related to the down-regulation of bcl-2 / bax ratio and the destruction of cytoskeleton.