肠胃清对裸小鼠大肠癌皮下移植瘤生长及STAT3、Bcl-2剪接异构体表达的影响  

Effects of Changweiqing(肠胃清)on Transplanted Tumor Growth of Colorectal Cancer Cells and Expression of STAT3 and Bcl-2 Gene Splicing Isoforms

作　　者：陈彬[1,2] 邓皖利 梁芳[1] 袁旭[1] 谢曼丽[1] 刘慧 CHEN Bin;DENG Wanli;LIANG Fang;YUAN Xu;XIE Manli;LIU Hui(Putuo Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai,200062;Shanghai University of Traditional Chinese Medicine)

机构地区：[1]上海中医药大学附属普陀医院,上海市普陀区200062 [2]上海中医药大学

出　　处：《中医杂志》2024年第8期849-857,共9页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81703887);上海市卫生健康委员会中医药科研项目(2020JP004);上海市普陀区卫生健康系统临床特色专科建设计划2021tszk01。

摘　　要：目的探讨肠胃清治疗大肠癌的可能作用机制。方法选用HCT 116肠癌细胞株,制备沉默多聚嘧啶区结合蛋白3(PTBP3)基因、过表达PTBP3基因稳转的肠癌细胞;72只裸小鼠随机分为3组,各组分别通过腋下注射沉默PTBP3的稳转细胞、过表达PTBP3的稳转细胞、未转染的肠癌细胞,构建沉默模型、过表达模型、对照模型3种大肠癌细胞皮下移植瘤模型,每种模型24只。造模成功后将对照模型裸小鼠随机分为对照模型组、对照肠胃清组、对照奥沙利铂组,沉默模型裸小鼠随机分为沉默模型组、沉默肠胃清组、沉默奥沙利铂组,过表达模型裸小鼠随机分为过表达模型组、过表达肠胃清组、过表达奥沙利铂组,每组均为8只。各肠胃清组肠胃清口服液灌胃剂量为35.9625 g/kg,腹腔注射0.2 ml生理盐水;各模型组以0.5 ml生理盐水灌胃,腹腔注射0.2 ml生理盐水;各奥沙利铂组腹腔注射奥沙利铂5 mg/kg(0.2 ml),以0.5 ml生理盐水灌胃,各组均灌胃每日1次,腹腔注射每周3次。给药31天后检测各组皮下瘤瘤重并计算抑瘤率,免疫组化法检测增殖细胞核抗原Ki67表达水平、细胞凋亡水平;实时荧光定量法、Western Blot检测各组皮下瘤组织PTBP3、信号转导及转录活化因子3(STAT3)剪接异构体α(STAT3α)、STAT3剪接异构体β(STAT3β)、B细胞淋巴瘤/白血病2(Bcl-2)剪接异构体α(Bcl-2α)、Bcl-2剪接异构体β(Bcl-2β)的mRNA及蛋白表达。结果在对照模型、沉默模型和过表达模型中,各肠胃清组和奥沙利铂组皮下瘤瘤重、皮下瘤组织Ki67表达水平均低于各相应模型组,细胞凋亡水平均高于各相应模型组(P<0.05或P<0.01);沉默肠胃清组和过表达肠胃清组皮下瘤组织PTBP3、STAT3α、Bcl-2α的mRNA和蛋白表达水平均低于相应模型组,STAT3β、Bcl-2β的mRNA和蛋白表达水平均高于相应模型组(P<0.05或P<0.01)。沉默各组皮下瘤瘤重、皮下瘤组织Ki67表达水平及皮下瘤组Objective To explore the possible mechanism of Changweiqing(肠胃清)in the treatment of colorectal cancer.Methods HCT 116 cancer cells were used to prepare intestinal cancer cells with silenced polypyrimidine region binding protein 3(PTBP3)gene and stably transfected cells with overexpressed PTBP3 gene.Stably transfected cells with silenced PTBP3,stably transfected cells with overexpressed PTBP3 and untransfected cancer cells were injected into the armpit of 72 nude mice to construct three different subcutaneous transplanted tumor models of colorectal cancer cells,including the silenced model,the overexpressed model and the control model,with 24 mice per model.Mice of each transplanted tumor modelwere randomly divided into Changweiqing(CWQ)group,oxaliplatin(OXA)group and normal saline(NS)group,with 8 mice in each group.The CWQ groups were given intragastric administra⁃tion of 35.9625 g/kg of Changweiqing oral liquid and were intraperitoneally injected with 0.2ml of normal saline;the NS groups were given 0.5ml of normal saline by gavage,and intraperitoneal injection of 0.2ml of normal saline;the OXA groups were intraperitoneally injected with 5 mg/kg(0.2 ml)of oxaliplatin and given 0.5ml of normal saline by gavage.Each group was given intragastric administration once a day and intraperitoneal injection three times a week.After 31 days,the weight of subcutaneous tumors in each group was measured,and the tumor inhibition rate of the groups in each model were measured.Immunohistochemistry and other methods were used to detect the expression level of cell proliferation cell nuclear antigen Ki67 and apoptosis index.Real-time PCR and Western Blot were used to detect mRNA and protein expressions of PTBP3,signal transducer and activator of transcription 3(STAT3)splicing isoformα(STAT3α),STAT3 splicing isoformβ(STAT3β),B-cell lymphoma/leukemia-2(Bcl-2)splicing isoformα(Bcl-2α),and Bcl-2 splicing isoformβ(Bcl-2β)in subcutaneous tumor cells in each group.Results For all three transplanted tumor models,the weight

关 键 词：大肠癌 选择性剪接 肠胃清 多聚嘧啶区结合蛋白3 信号转导及转录活化因子3 B细胞淋巴瘤/白血病2 

分 类 号：R285.5[医药卫生—中药学]