健脾解毒方逆转大肠癌细胞多药耐药的作用机制  

Mechanism of multidrug resistance of colon cancer cell reversed by Jianpijiedu Herb

作　　者：李先茜[1] 吴嘉熙[1] 范忠泽[2] 孙燕妮[2] 高虹[2] 

机构地区：[1]上海市徐汇区中心医院,上海200031 [2]上海中医药大学附属普陀医院

出　　处：《山东医药》2014年第33期13-16,共4页Shandong Medical Journal

基　　金：上海市卫生局科研课题(2006116)

摘　　要：目的探讨中医健脾解毒方逆转大肠癌多药耐药(MDR)的机制。方法采用细胞生长抑制实验观察长春新碱在健脾解毒方治疗前后对大肠癌耐药细胞株(HCT-8/V)敏感性的改变,实时荧光定量PCR及Western blot方法观察健脾解毒方对大肠癌细胞Akt磷酸化程度及MDR1基因表达的影响。结果健脾解毒方药物血清与大肠癌细胞培养24、48、72 h后,长春新碱对细胞的生长抑制率分别为5.9%、11.0%、15.0%,明显高于空白对照组(P均<0.05),其抑制作用与健脾解毒方的药物血清浓度及作用时间呈正相关。健脾解毒方血清能有效降低HCT-8/V中MDR-1的蛋白及mRNA表达(P<0.05);健脾解毒方逆转MDR-1的表达与其所致的细胞内Akt磷酸化水平下降呈正相关,与长春新碱敏感性提高呈线性负相关。健脾解毒方血清与PI3K抑制剂LY294002联用能协同降低HCT-8/V中MDR-1的表达水平和细胞内Akt磷酸化程度,较单用更能增加长春新碱的敏感性。结论健脾解毒方能有效减低耐药肿瘤细胞内Akt磷酸化,从而降低PI3K/Akt信号通路活性,后者进一步导致MDR1表达下降,从而增加长春新碱对大肠癌细胞的敏感性。Objective To investigate the mechanism of which Chinese traditional medicine , Jianpijiedu （ JBJD） herb protocol reverses multidrug resistance （ MDR） of colon cancer .Methods Sensitivity of Vincristine to colon cancer cells （ HCT-8/V） were detected by cell growth inhibition metods befor or after the treatment of Jianpijiedu Herb .Expression of multidrug gene 1 （ MDR-1） and phosphorylation of Akt in cancer cell were studied by either real -time PCR or Western blot.Results After HCT-8/V was co-cultured with serum containing 20% of JBJD for 24 h, 48 h and 72 h, inhibition rates on proliferation of HCT-8/V resulted by Vincristine were 5.9%, 11.0% and 15.0%, respectively, and all higher than that of the blank control group （all P〈0.05）.Inhibiting cell growth of Vincristine was positively correlated to the co-culture time and the serum concentration of JBJD .JBJD down-regulated the expression of MDR-1 protein and mRNA in HCT-8/V（P〈0.05）.Expression of MDR-1 was positively related with the decreased degree of Akt phosphorylation induced by JBJD, and negatively correlated with sensitivity of Vincristine .JBJD combined with PI3K inhibitor（LY294002） resulted in significant deduction of MDR-1 expression at mRNA and protein levels （P-gp） and Akt phosphorylation （p-Akt）, and enhanced Vincristine sensitivity significantly than that by single use of JBJD or LY 294002 .Conclusion JBJD effectively turn down activity of PI 3K/Akt signaling pathways by deduction of cellular Akt phosphorylation , which down-regulate the expression of MDR1, increase the sensitivity of Vincristine .

关 键 词：大肠肿瘤 健脾解毒方 多药耐药基因 PI3 K Akt信号因子 

分 类 号：R735.3[医药卫生—肿瘤]