雷帕霉素、RAD001抑制胃癌细胞株MKN-28、MKN-45细胞生长的实验研究

An experimental study of the inhibitory effect of rapamycin and RAD001 on the growth of gastric carcinoma cell lines MKN-28 and MKN-45

目的探讨雷帕霉素(rapamycin)、RAD001(everolimus)对胃癌细胞株MKN-28、MKN-45细胞生长的影响,并对其机制进行初步探讨。方法体外培养人胃癌MKN-28、MKN-45细胞,给予雷帕霉素、RAD001干预后通过细胞计数、流式细胞术检测肿瘤细胞生长、细胞周期分布情况。结果雷帕霉素、RAD001均可抑制MKN-28、MKN-45细胞生长,抑制作用具有时间依赖性,雷帕霉素作用于MKN-28、MKN-45两种细胞株72 h抑制率分别为23.40%、36.26%;RAD001作用于MKN-28、MKN-45两种细胞株72 h抑制率分别为25.12%、40.11%。72 h时RAD001组与阴性对照组、DMSO对照组相比,MKN-28、MKN-45细胞计数差异有显著性(P<0.05)。RAD001作用24 h后MKN-28、MKN-45细胞周期发生改变,停滞在G0-G1期细胞比例增加。RAD001组与阴性对照组、DMSO对照组相比,MKN-45细胞周期差异有显著性(P<0.05),MKN-28细胞周期差异无显著性(P>0.05)。结论两种药物相比较,RAD001相对敏感;两株细胞相比较,MKN-45相对敏感。RAD001通过调节细胞周期进而抑制胃癌细胞的增殖。

Objective To investigate the effect of rapamycin and RAD001 on the growth of gastric carcinoma cell lines MKN -28 and MKN- 45 and to investigate its possible mechanism. Methods Gastric carcinoma cell lines MKN -28 and MKN -45 cultured in vitro were treated by rapamycin and RADO01 （20 nmol/L）, and then the cell Coulter Counter and flow cytometry were used to detect cell growth and cell cycle. Results Both rapamycin and RAD001 could inhibit the growth of gastric carcinoma cell lines MKN - 28 and MKN - 45 in a time - dependence manner. The inhibition percentage of rapamyein at G2 phase （72 h） was 23.40% and 36.26% , and that of RAD001 at G2 phase was 25.12% and 40.11%, respectively. The cell counts of MKN -28 and MKN -45 in RAD001 group had significant differences compared with those in the negative control group and DMSO control group （ P 〈 0.05 ）. At Go phase or 24 h after RADO01 treatment, the cell cycles of MKN - 28 and MKN - 45 changed and the proportion of cells stagnating at G0 - G1 increased. In RADO01 group, the difference in MKN - 45 cell cycle was significant （ P 〈 0. 05 ） , while that in MKN - 28 cell cycle was insignificant （ P 〉 0.05 ） , as compared to the negative control and DMSO control groups. Conclusion Of the two drugs, RAD001 is more sensitive to MKN -28 and MKN -45, while of the two cell lines, MKN-45 is more sensitive to rapamycin and RAD001. The result suggests that RAD001 can inhibit the growth of gastric carcinoma cells by its adiustment to cell cycles.