健脾消积方水提物对人肝癌SMMC7721细胞自噬的调控机制  

Regulatory effect of water extract of Jianpi Xiaoji prescription on autophagy of human liver cancer SMMC7721 cells

作　　者：韩志伟 王宗玉 鞠佳霓 李科志[1] 邬国斌[1] 林栋毅 陈闯[1] HAN Zhiwei;WANG Zongyu;JU Jiani(Guangxi Medical University Cancer Hospital,Nanning 530021,China)

机构地区：[1]广西医科大学附属肿瘤医院,南宁530021 [2]中山大学孙逸仙纪念医院,广州510030

出　　处：《临床肝胆病杂志》2020年第3期587-591,共5页Journal of Clinical Hepatology

基　　金：国家自然科学基金项目(81560726);广西科技计划项目(AB18126066);中央引导地方科技发展专项(桂科ZY1949017);南宁市科学研究与技术开发计划(20193116)。

摘　　要：目的探讨健脾消积方水提物抗肝癌机制以及对细胞自噬流和自噬相关蛋白的影响。方法提取健脾消积方水提物,作用于SMMC7721细胞,使用CCK-8法检测细胞增殖,流式细胞学检测细胞的凋亡率,使用Western Blot检测自噬相关蛋白Beclin1、LC3和P62的表达,并监测自噬体的形成及自噬体与溶酶体的融合过程。计量资料两组比较采用独立样本t检验,多组比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果CCK8检测显示,细胞加药后24 h、48 h、72 h、96 h,增殖均受到了明显抑制,与对照组相比差异均有统计学意义(t值分别为28.458、81.093、85.328、100.158,P值均<0.001)。流式细胞学凋亡检测结果显示,加药组肝癌SMMC7721细胞凋亡较对照组显著增加(t=-42.629,P<0.001)。Western Blot结果显示,与对照组相比,加药组Beclin1蛋白表达下调,自噬标志蛋白LC3Ⅱ表达上调,P62表达上调。加药组细胞的自噬体向自噬溶酶体的流动受到了阻滞,与对照组相比,差异有统计学意义(F=31.155,P<0.001)。结论健脾消积方水提物能够抑制人肝癌SMMC-7721细胞的增殖,促进细胞凋亡;同时能上调Beclin1的表达,下调LC3和P62的表达并阻滞自噬流的形成。Objective To investigate the anti-liver cancer mechanism of water extract of Jianpi Xiaoji prescription and its effect on cell autophagy flow and autophagy-related proteins. Methods SMMC7721 cells were treated with water extract of Jianpi Xiaoji prescription.CCK-8 assay was used to measure cell proliferation;flow cytometry was used to measure cell apoptosis rate;Western blot was used to measure the expression of the autophagy-related proteins Beclin1,LC3,and P62;the formation of autophagosomes and the fusion of autophagosomes and lysosomes were monitored. The independent samples t-test was used for comparison of continuous data between two groups;a one-way analysis of variance was used for comparison between multiple groups,and the least significant difference t-test was used for further comparison between two groups. Results The CCK-8 assay showed that cell proliferation was significantly inhibited at 24,48,72,and 96 hours after the drug was added,and there was a significant difference between the drug treatment group and the control group( t =28. 458,81. 093,85. 328,and 100. 158,all P < 0. 001). Flow cytometry showed that compared with the control group,the drug treatment group had a significant increase in the apoptosis of liver cancer SMMC7721 cells( t =-42. 629,P < 0. 001). Western Blot showed that compared with the control group,the drug treatment group had significantly downregulated expression of Beclin1 and significantly upregulated expression of the autophagy markers LC3-Ⅱ and P62. The flow of autophagosomes to autolysosomes was blocked in the drug treatment group,and there was a significant difference between the drug treatment group and the control group( F = 31. 155,P < 0. 001). Conclusion The water extract of Jianpi Xiaoji prescription can inhibit the proliferation of human liver cancer SMMC-7721 cells and promote apoptosis;meanwhile,it can upregulate the expression of Beclin1,downregulate the expression of LC3 and P62,and block the formation of autophagy flux.

关 键 词：肝肿瘤 自噬 健脾消积方 

分 类 号：R735.7[医药卫生—肿瘤]