伪士的宁对人结肠癌HT-29细胞的增殖及细胞周期的影响

Study on the effect of pseudo-strychnine on proliferation and cell cycle of human colon cancer HT-29 cells

目的研究伪士的宁对人结肠癌HT-29细胞的增殖及细胞周期的影响。方法通过MTT法比较在31.25μmol/L、62.50μmol/L、125μmol/L、250μmol/L、500μmol/L、1000μmol/L共6个浓度的伪士的宁培养24小时、48小时、72小时后对HT-29细胞增殖的抑制作用;通过显微镜细胞观察法检测细胞生长状态;利用流式细胞术检测250μmol/L伪士的宁对HT-29细胞周期的影响。结果伪士的宁对人结肠癌HT-29细胞有一定的抑制作用,其抑制作用与浓度和作用时间呈正相关,1000μmol/L培养72小时作用最显著,抑制率为97.47%;随着伪士的宁浓度增加,作用时间增大,HT-29细胞密度逐渐变小,局部可见固缩细胞或死亡细胞。伪士的宁(250μmol/L)作用于HT-29细胞后,与空白组相比,G1期细胞数量明显增加,S期细胞数量不变,G2期细胞数量减少,差异有统计学意义(P<0.05),表明伪士的宁可诱使HT-29细胞在G1期发生阻滞,诱导细胞凋亡。结论伪士的宁对人结肠癌HT-29细胞增殖具有显著抑制作用,且其作用呈现时间依赖和剂量依赖关系,其抑制作用机制可能与阻滞细胞周期有关。

Objective To study the effect of pseudo-strychnine on proliferation and cycle of human colon cancer HT-29 cells.Methods MTT method was used to compare the inhibitory effect of 6 concentrations(31.25μmol/L,62.50μmol/L,125μmol/L,250μmol/L,500μmol/L and 1000μmol/L)of pseudo-strychnine on the proliferation of HT-29 cells after 24h,48h and 72h of culture.The cells growth state was observed by microscope;Flow cytometry were used to analyze the effect of 250μmol/L pseudo-strychnine on HT-29 cells’cell cycle.Results Pseudo-strychnine has some inhibitory effect on human colon cancer HT-29 cells,and its inhibitory effect has a positive relationship with concentration and time,the most significant effect concentrations was 1000μmol/L after being cultured 72h,the inhibitory rate was 97.47%.The density of HT-29 cells decreased and partial pyknosis cells or dead cells can be observed locally,gradually with the increasing pseudo-strychnine concentration and exposure time.Compared with the blank group,the number of cells in G1 phase increased significantly,the number of cells in S phase remained unchanged and the number of cells in G2 phase decreased after treated with pseudo-stigine(250μmol/L),the difference was statistically significant(P<0.05).HT-29 cells was arrested at the G 1 phase after pseudo-strychnine intervention to induce cell apoptosis.Conclusion Pseudo-strychnine has significant inhibitory effect on human colon cancer HT-29 cells,and has dose-dependent and time-dependent the effect,the mechanism of inhibitory effect may be related to the arrest of cell cycle.