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目的 探讨黄连素对结肠癌细胞系HT 2 9的作用及其相关机制 ,为阐明黄连素作为一种新的结肠癌化学治疗药物进行理论上的准备并提供相关实验结果。方法 黄连素 0 1、0 3、3 0、30 0 μmol/L加入到HT 2 9结肠癌细胞系培养液中 ,每天测量各有关值。同时以NS 398为环氧合酶(COX 2 )活性抑制剂 ,对比观察黄连素对COX 2活性的影响。采用氮蓝四唑盐实验法检测细胞的生长和增殖 ,逆转录 PCR法检测COX 2mRNA ,免疫细胞化学法检测COX 2蛋白表达 ,ELISA法检测前列腺素 (PG)E2 的含量。结果 黄连素在浓度大于 0 3μmol/L时则有明显的量效关系。黄连素在浓度大于 0 3μmol/L时对COX 2mRNA水平和蛋白的表达有明显的抑制作用。黄连素对PGE2 的生成有显著抑制作用。结论 黄连素抑制COX 2在mRNA水平和蛋白质水平的表达 ,同时也抑制COX 2活性 ,进而抑制PGE2 的生成 ,这可能为黄连素抑制HT 2 9细胞生长和增殖的机制之一。
Objective To investigate the effect and mechanism of berberine on human colon cancer cell line HT 2 9 in order to elucidate the theoretical preparation of berberine as a new chemotherapeutic agent for colon cancer and to provide relevant experimental results. Method Berberine 0 1,03 0,30 0 μmol / L was added to HT 2 9 colon cancer cell line culture medium, and the relevant values were measured daily. At the same time, NS 398 was used as cyclooxygenase (COX 2) inhibitor to compare the effect of berberine on COX 2 activity. The cell growth and proliferation were detected by the method of nitrogen blue tetrazolium salt assay. COX 2 mRNA was detected by RT-PCR, the expression of COX 2 protein by immunocytochemical method and the content of prostaglandin (PG) E2 by ELISA. Results berberine at a concentration greater than 0 3μmol / L when there is a significant dose-effect relationship. Berberine significantly inhibited COX 2 mRNA and protein expression at a concentration of greater than 0 3 μmol / L. Berberine has a significant inhibitory effect on the production of PGE2. Conclusion Berberine inhibits the expression of COX 2 at mRNA and protein levels as well as inhibits COX 2 activity and thus PGE 2 production. This may be one of the mechanisms by which berberine inhibits the growth and proliferation of HT 2 9 cells.