基于PI3K-Akt信号通路的清带汤对大肠癌细胞增殖的影响  

Effects of Qingdai Decoction on Colorectal Cancer Proliferation Based on PI3K-Akt Signaling Pathway

作　　者：陈彦华 陈佳旻[3] 刘蕾 王连洁 吴范晨 冯媛媛 周利红[1] CHEN Yanhua;CHEN Jiamin;LIU Lei;WANG Lianjie;WU Fanchen;FENG Yuanyuan;ZHOU Lihong(Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Academy of Integrated Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Punan Hospital,Shanghai 200125,China)

机构地区：[1]上海中医药大学附属曙光医院,上海201203 [2]上海中医药大学中西医结合研究院,上海201203 [3]上海浦南医院,上海200125

出　　处：《中国中医药信息杂志》2023年第9期88-95,共8页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81973651、82174145);上海市中医药三年行动计划[ZY(2021-2023)-0208];上海市炎癌转化病症生物学前沿研究基地(2021KJ03-12)。

摘　　要：目的通过体内外实验观察清带汤对大肠癌细胞增殖的抑制作用,并通过网络药理学、分子对接技术和实验探讨其可能的机制。方法体外培养人大肠癌HCT116、LoVo、HT29细胞和正常人肠上皮NCM460细胞,加入不同浓度(10、20、40、80、160μg/mL)清带汤冻干粉干预48 h,CCK-8法检测细胞增殖;建立人大肠癌HCT116细胞荷瘤裸鼠模型,将裸鼠随机分为对照组和清带汤组(11.6 g/kg),给药21 d后观察裸鼠瘤体生长情况;采用网络药理学预测清带汤治疗大肠癌有效成分、核心靶点及相关通路,利用分子对接技术分析清带汤有效成分与核心靶点的结合能力;Western blot检测清带汤干预后HCT116和LoVo细胞PI3K-Akt信号通路相关蛋白表达,q PCR检测HCT116、LoVo细胞及裸鼠肿瘤组织PI3K-Akt信号通路相关基因表达。结果清带汤(20、40、80、160μg/mL)对HCT116、LoVo、HT29细胞增殖有显著抑制作用(P<0.05,P<0.01),而对NCM460细胞无明显影响。与对照组比较,清带汤组裸鼠皮下移植瘤体积和质量显著降低(P<0.05)。网络药理学和分子对接结果显示,清带汤抑制大肠癌可能与PI3K-Akt信号通路有关。Western blot结果显示,与对照组比较,清带汤组HCT116、LoVo细胞p-AKT和p-mTOR蛋白表达均明显降低(P<0.05,P<0.01);qPCR结果显示,与对照组比较,清带汤组HCT116、LoVo细胞及裸鼠肿瘤组织PI3K-Akt信号通路下游基因CDK2、BCL-2和MCL-1表达均显著降低(P<0.05,P<0.01)。结论清带汤能有效抑制人大肠癌细胞增殖及荷瘤裸鼠瘤体生长,其机制与调控PI3K-Akt信号通路、抑制癌细胞增殖有关。Objective To observe the inhibitory effects of Qingdai Decoction on colorectal cancer cells through in vivo and in vitro experiments;To explore its possible mechanism by network pharmacology,molecular docking,and experiment.Methods HCT116,LoVo,HT29 and NCM460 cells were cultured in vitro,and different concentrations of Qingdai Decoction(10,20,40,80,160μg/mL)were added for 48 h respectively.The cell proliferation was detected by CCK-8 assay.The nude mouse model of HCT116 cells was established,tumor-forming nude mice were randomly divided into model group and Qingdai Decoction group(11.6 g/kg),tumor growth in nude mice after 21 days was observed.The network pharmacology method was used to predict the effective components,core targets and related pathways of Qingdai Decoction in the treatment of colorectal cancer.The binding ability of the main components of Qingdai Decoction to the core targets was analyzed by molecular docking technique.Western blot was used to detect the expression of PI3K-Akt signaling pathway related proteins in HCT116 and LoVo cells after intervention with Qingdai Decoction;qPCR was used to detect the expression of PI3K-Akt signaling pathway related genes in HCT116,LoVo cells and nude mouse tumor tissue.Results Qingdai Decoction(20,40,80,160μg/mL)had a significant inhibitory effect on the proliferation of HCT116,LoVo and HT29 cells(P<0.05,P<0.01),while NCM460 cells were not affected.Compared with the control group,the volume and mass of subcutaneous transplanted tumors in nude mice in the Qingdai Decoction group were significantly reduced(P<0.05).The results of network pharmacology and molecular docking indicate that the inhibition of colorectal cancer by Qingdai Decoction may be related to the PI3K-Akt signaling pathway.Western blot results showed that compared with the control group,the expressions of p-AKT and p-mTOR proteins in HCT116,LoVo cells of the Qingdai Decoction group significantly reduced(P<0.05,P<0.01);the qPCR results showed that compared with the control group,the expressions

关 键 词：大肠癌 清带汤 网络药理学 分子对接 PI3K-AKT信号通路 

分 类 号：R285.5[医药卫生—中药学]