基于PI3K/Akt信号通路探讨重楼皂苷Ⅶ对人结直肠癌细胞增殖的影响  

Effects of PolyphyllinⅦon Proliferation of Colorectal Cancer Cells Based on PI3K/Akt Signaling Pathway

作　　者：侯亚妮 孙中华 刘志勇 褚志杰[2] 白克运[2] HOU Yani;SUN Zhonghua;LIU Zhiyong;CHU Zhijie;BAI Keyun(Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,China)

机构地区：[1]山东中医药大学,山东济南250355 [2]山东中医药大学附属医院,山东济南250014

出　　处：《山东中医药大学学报》2023年第3期330-335,384,共7页Journal of Shandong University of Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(编号:82104916);山东省自然科学基金青年项目(编号:ZR2020QH337)。

摘　　要：目的:观察重楼皂苷(PP)Ⅶ对人结直肠癌HT29和LoVo细胞增殖的影响及可能机制。方法:使用梯度浓度的PPⅠ、PPⅡ、PPⅦ处理人结直肠癌HT29和LoVo细胞,四甲基偶氮唑盐(MTT)比色法检测药物对细胞活力的影响,统计药物对应最大半抑制浓度(IC50)值;克隆形成实验检测PPⅦ干预10 d对人结直肠癌HT29和LoVo细胞克隆形成的影响;细胞增殖检测实验分析梯度浓度PPⅦ作用48 h对人结直肠癌HT29和LoVo细胞增殖的影响;通过蛋白质印迹法(Western blotting)检测HT29和LoVo细胞中磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt)通路中内PI3K、磷酸化磷脂酰肌醇3激酶(p-PI3K)、Akt、磷酸化蛋白激酶B(p-Akt)的蛋白水平变化。结果:PPⅠ、PPⅡ、PPⅦ均能降低HT29和LoVo细胞的活力,以PPⅦ效果最为显著;PPⅦ能够有效抑制HT29和LoVo细胞的克隆形成能力;PPⅦ能够抑制HT29和LoVo细胞的增殖能力。PPⅦ各剂量组可降低对应的p-PI3K、p-Akt蛋白表达水平。结论:PPⅦ可能通过下调PI3K和Akt的磷酸化水平,抑制PI3K/Akt信号通路,从而抑制结直肠癌细胞的增殖。Objective:To investigate the effects and possible mechanism of polyphyllin(PP)Ⅶon proliferation of human colorectal cancer HT29 and LoVo cells.Methods:The effects of gradient concentrations of PPⅠ,PPⅡ,PPⅦon cell viability were detected by methyl thiazolyl tetrazolium(MTT)method,and the maximum inhibition concentration(IC50)values of drugs were statistically calculated.The number of clonal formations were assayed by clonal formation experiments after PPⅦwas added to HT29 and LoVo cells and treated for 10 days.The cell proliferation assay was used to assess the effects of PPⅦon cell proliferation after 48 h of intervention on HT29 and LoVo cells.The expression levels of proteins in phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)pathway,including PI3K,phosphorylated PI3K(p-PI3K),Akt and phosphorylated Akt(p-Akt)were evaluated by Western blot.Results:PPⅠ,PPⅡand PPⅦcould decrease the activity of HT29 and LoVo cells,and the effect of PPⅦwas the most significant.PPⅦcould effectively suppress the cloning formations of HT29 and LoVo cells.PPⅦcould effectively inhibit the proliferation ability of HT29 and LoVo cells.The expression levels of p-PI3K and p-Akt proteins in each dose group of PPⅦwere reduced.Conclusion:PPⅦmay inhibit the proliferation of colorectal cancer cells by down-regulating the phosphorylation of PI3K and Akt and inhibiting PI3K/Akt signaling pathway.

关 键 词：重楼皂苷Ⅶ 结直肠癌 细胞增殖 磷脂酰肌醇3激酶/蛋白激酶B信号通路 人结直肠癌HT29细胞 人结直肠癌LoVo细胞 

分 类 号：R285.5[医药卫生—中药学]