健脾消癌方对结肠癌细胞中Akt/mTOR信号通路的影响

Effect of Jianpi Xiao’ ai Prescription on Akt/mTOR Signaling Pathways in Colorectal Cells

目的:探讨健脾消癌方含药血清对人结肠癌HCT116细胞蛋白激酶B/哺乳动物雷帕霉素靶蛋白(Akt/mTOR)信号通路相关蛋白表达的影响。方法:以15%健脾消癌方含药血清处理结肠癌HCT116细胞,采用Transwell侵袭实验检测HCT116细胞迁移、侵袭作用,采用蛋白质印迹法(Western blot)检测HCT116细胞中Akt,磷酸化蛋白激酶B(p-Akt),mTOR,磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR),核糖体S6蛋白激酶(S6K1),磷酸化核糖体S6蛋白激酶(p-S6K1),真核细胞始动因子4E结合蛋白(4EBP1),磷酸化真核细胞始动因子4E结合蛋白(p-4EBP1)等蛋白的表达,正常血清组设立相同浓度正常血清组(15%),10%胎牛血清组(FBS)。结果:与正常血清组比较,健脾消癌方含药血清组迁移细胞数与侵袭细胞数均显著降低(P<0.01);与正常血清组比较,Akt蛋白表达无明显下调,p-Akt蛋白表达显著下调(P<0.01);与正常血清组比较,mTOR蛋白表达无明显下调,p-mTOR蛋白表达显著下调(P<0.01);与正常血清组比较,S6K1蛋白表达无明显下调,p-S6K1蛋白表达显著下调(P<0.01);与正常血清组比较,4EBP1蛋白表达无明显下调,p-4EBP1蛋白表达显著下调(P<0.01)。结论:健脾消癌方抗结肠癌复发转移的机制可能与抑制Akt/mTOR信号通路的激活有关。

Objective:To investigate the effect of drug-containing serum of Jianpi Xiaoai prescription on protein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathways in colorectal cells HCT116.Method:The HTC116 cells were treated by 15%concentration of drug-contained serum,and then the cell migration and invasion were detected by Transwell assay,the protein expression levels of Akt,phosphorylated protein kinase B(p-Akt),mTOR,phosphorylated mammalian target of rapamycin(p-mTOR),ribosomal protein S6 kinase,polypeptide1(S6 K1),phosphorylated ribosomal protein S6 kinase,polypeptide1(p-S6 K1),4 E-binding protein1(4 EBP1),and phosphorylated 4 E-binding protein1(p-4 EBP1)in HCT116 cells were detected by Western blot.The control group was treated by untreated serum(15%),and 10%fetal bovine serum(FBS).Result:As compared with the control group,the number of migration and invasion cells was significantly reduced in drug-contained serum group(P<0.01),the expression of Akt had no obvious decrease,p-Akt protein expression was significantly lowered in the drug-contained serum group(P<0.01),the expression of mTOR had no obvious decrease,but p-mTOR protein expression was significantly lowered in drug-contained serum group(P<0.01),the expression of S6 K1 had no obvious decrease,but p-S6 K1 protein expression was significantly lowered in the drug-contained serum group(P<0.01),the protein expression of 4 EBP1 had no obvious decrease,but p-4 EBP1 protein expression was significantly lowered in the drug-contained serum group(P<0.01).Conclusion:The anti-tumor mechanism and transfer of Jianpi Xiaoai prescription may be related to inhibiting the activation of Akt/mTOR signaling pathways in colorectal cancer.