摘要: |
目的:探讨健脾消癌方(JPXA)含药血清对人结肠癌HCT116细胞丝氨酸/苏氨酸蛋白激酶(AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)表达以及对细胞增殖、细胞周期及凋亡的影响。方法:取不同浓度的JPXA含药血清作用于HCT116细胞,筛选JPXA含药血清处理HCT116细胞的半抑制浓度(IC50),并将HCT116细胞分为实验组和对照组,予以相应处理后,采用CCK-8法检测细胞增殖能力,PI-FACS法检测细胞周期变化,Annexin V-APC 单染法流式细胞仪检测凋亡水平,并采用Rt-PCR检测AKT mRNA、mTOR mRNA水平,采用Western Blot检测AKT、p-AKT、mTOR及磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)水平。结果:与对照组相比,实验组HCT116细胞增殖能力显著下降,能使细胞周期停留在G1期及S期,且细胞凋亡水平上升,差异均有统计学意义(P<0.05)。与对照组相比,实验组AKT mRNA、mTOR mRNA表达水平及AKT、p-AKT、mTOR、p-mTOR蛋白含量明显下降(P<0.05或P<0.01)。结论:JPXA能抑制AKT、mTOR表达,对结肠癌细胞的增殖、克隆能力有一定抑制作用,并可在一定程度上促进结肠癌细胞的凋亡。 |
关键词: 结肠癌 健脾消癌方 HCT116 AKT mTOR 实验研究 |
DOI: |
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Effect of Jianpi Xiaoai prescription on the expression of protein kinase B and mammalian target of rapamycin in human colon cancer HCT116 cells |
REN Lizhi,SONG Cheng,TANG Wei |
(Hunan Cancer Hospital & The Affiliated Tumor Hospital,Xiangya School of Medicine,Central South University,Changsha 410013,Hunan,China;The Affiliated Hospital of Hunan Academy of Chinese Medicine,Changsha 410006,Hunan,China) |
Abstract: |
Objective:To investigate the effect of serum containing Jianpi Xiaoai prescription (JPXA) on the expression of serine/threonine protein kinase B (AKT) and mammalian target of rapamycin (mTOR) in human colon cancer HCT116 cells and the proliferation,cell cycle,and apoptosis of HCT116 cells.Methods:HCT116 cells were treated with different concentrations of serum containing JPXA to screen out the half-maximal inhibitory concentration of serum containing JPXA in the treatment of HCT116 cells.HCT116 cells were divided into experimental group and control group;after corresponding treatment,CCK-8 assay was used to measure cell proliferative capacity,PI-FACS was used to measure the change in cell cycle,Annexin V-APC single-staining flow cytometry was used to measure the level of cell apoptosis,RT-PCR was used to measure the mRNA expression levels of AKT and mTOR,and Western blot was used to measure the protein expression levels of AKT,phosphorylated AKT (p-AKT),mTOR,and phosphorylated mTOR (p-mTOR).Results:Compared with the control group,the experimental group had a significant reduction in the proliferative capacity of HCT116 cells,with most cells arrested in G1/S phase,and a significant increase in the level of cell apoptosis (P<0.05).Compared with the control group,the experimental group had significant reductions in the mRNA expression levels of AKT and mTOR and the protein expression levels of AKT,p-AKT,mTOR,and p-mTOR (P<0.05 or P<0.01).Conclusion:JPXA can inhibit the expression of AKT and mTOR and the proliferation and clone abilities of colon cancer cells,and it can also promote the apoptosis of colon cancer cells to a certain extent. |
Key words: colon cancer Jianpi Xiaoai prescription HCT116 protein kinase B mammalian target of rapamycin experimental study |