姜黄素对结肠癌HT-29细胞生长增殖、细胞周期分布及CyclinD1基因表达的影响

Influence of curcumin on proliferation,cell cycle distribution of HT-29 cells and CyclinD1 gene expression

目的观察姜黄素对人结肠癌HT-29细胞生长增殖、细胞周期分布的影响,并探讨可能的机制。方法培养对数生长的人结肠癌HT-29细胞,用不同浓度的姜黄素-1640培养基进行培养。选取24、48、72h时间节点,观察细胞生长的形态学差异。四甲基偶氮唑蓝(MTT)法检测姜黄素对HT-29细胞的生存抑制率,流式细胞术检测加入姜黄素前后HT-29细胞的周期分布改变,并对药物处理前后的CyclinD1基因表达进行检测比较。结果在加入姜黄素稀释液后,HT-29细胞生长明显变慢,形状由饱满变得细长,脱壁增多。MTT提示姜黄素能明显抑制HT-29细胞生长,并呈时间和浓度依赖性。流式细胞术显示姜黄素能诱导明显的G0/G1期阻滞,24、48、72h时G1期比例分别为38.2%、44.6%、49.3%,相对对照组差异有统计学意义(P<0.05)。蛋白免疫印迹(Western blot)实验发现姜黄素能明显降低HT-29细胞中的CyclinD1基因表达,并随作用时间的延长,抑制作用更强。结论姜黄素能明显抑制人结肠癌HT-29细胞细胞周期CyclinD1基因表达,并诱导细胞G1期阻滞。

Objective To investigate the effect of curcumin on The proliferation and cell cycle distribution of human colon cancer cells HT-29 and its potential mechanism. Methods HT-29 cells were cultured in logarithmic growth phase with,different concentrations of curcumin-1640 medium. The morphologic differences of cellular growth were observed at the time points of 24, 48,72 h. The survival inhibition rate of curcumin on HT-29 cells was analyzed by MTT test. Then the changes of cell cycle distribu- tion before and after adding curcumin was tested by flow cytometry. Western blot assay was adopted to detect the expression change of CyclinD1 in HT-29 cells after treating by curcumin. Results After adding curcumin diluent, the HT-29 cells growth obviously turned to slow, the shape was changed from satiation to long and thin, pulling off from wall was increased. MTT indicated that cur- cumin could significantly inhibit HT-29 cells growth at＇a time- and dose-dependent manner. As showing by flow cytometry, curcu- min could induce Go^G1 arrested. The G1 phase proportions at 24,48,72 h were 38. 2%,44.6 % and 49. 3% respectively;the differ- ence compared with the control group was statistically significant. Western blot test found that curcumin could significantly decrease the CyclinD1 gene expression in HT-29 cells, moreover the inhibiting effect became stronger with the action time extending. Conclu- sion Curcumin can significantly inhibit the expression of CyclinD1 gene in human colon cancer cell line HT-29 and induces G1 phase arrest subsequently.