摘要
目的:以体外培养SW480细胞和体内荷瘤裸鼠为研究对象,探讨大黄酚(Chr)对结肠癌SW480细胞增殖、侵袭和裸鼠体内肿瘤形成的影响。方法:分别在结肠癌细胞SW480中加入25、50和100μmol/L大黄酚,Brdu染色法检测细胞增殖情况,Western blot检测增殖相关蛋白Ki67和PCNA表达水平;划痕实验和Transwell实验检测细胞迁移及侵袭能力;Western blot检测VEGF、E-cadherin和N-cadherin等上皮间充质转化相关蛋白表达,免疫荧光法检测波形蛋白表达情况;Western blot检测信号通路AMPK、mTOR和cyclin D1表达;建立移植瘤裸鼠模型,分别腹腔注射12.5、25和50 mg/kg大黄酚,检测荷瘤小鼠存活率、肿瘤重量,采用免疫组织化学法检测肿瘤组织中Ki67和PCNA表达水平,Western blot检测信号通路AMPK、mTOR和cyclin D1活化情况。结果:体外实验表明,大黄酚能剂量依赖性地抑制人结肠癌SW480细胞增殖和Ki67、PCNA蛋白表达(P<0.05或P<0.01),降低细胞划痕闭合率和侵袭细胞数量(P<0.05或P<0.01),调节E-cadherin、VEGF、N-cadherin、Vimentin、AMPK、mTOR和cyclin D1在SW480细胞中的表达水平(P<0.05或P<0.01);体内实验表明,与对照组(0.45±0.09)g相比,大黄酚各剂量组肿瘤组织重量分别为(0.32±0.06)g、(0.24±0.07)g和(0.15±0.04)g,差异具有统计学意义(P<0.05或P<0.01),且大黄酚剂量依赖性地抑制Ki67和PCNA表达(P<0.05或P<0.01),调控AMPK、p-mTOR和cyclin D1在肿瘤组织的表达(P<0.05或P<0.01)。结论:大黄酚通过AMPK信号通路抑制结肠癌SW480细胞的增殖、侵袭及荷瘤小鼠肿瘤形成。
Objective:Using SW480 cells cultured in vitro and tumor-bearing nude mice as research objects,effects of chrysophanol(Chr)on proliferation and invasion of colon cancer SW480 cells and tumor formation in nude mice were studied.Methods:Adding 25,50 and 100μmol/L Chr to SW480 cells,proliferation of cells was detected by Brdu staining,expressions of Ki67 and PCNA were detected by Western blot,migration and invasion of cells were detected by scratch and Transwell experiments,and expressions of VEGF,E-cadherin and N-cadherin were detected by Western blot,expression of Vimentin was detected by immunofluorescence,expression of AMPK,mTOR and cyclin D1 were detected by Western blot.Nude mice models of transplanted tumors were intraperitoneally injected with 12.5,25 and 50 mg/kg Chr respectively,survival rate of mice and tumor weight of tumor tissues were measured,expressions of Ki67 and PCNA in tumor tissues were detected by immunohistochemistry,and activation of signal pathways AMPK,mTOR and cyclin D1 were detected by Western blot.Results:In vitro experiments showed that Chr could inhibit proliferationand and expressions of Ki67 and PCNA in human colon cancer SW480 cells in a dose-dependent manner(P<0.05 or P<0.01),reduce scratch closure rate and number of invasive cells(P<0.05 or P<0.01)and regulate protein expressions of E-cadherin,VEGF,N-cadherin,Vimentin,AMPK,mTOR and cyclin D1 in SW480 cells(P<0.05 or P<0.01).In vivo experiments,compared with control group(0.45±0.09)g,weight of tumor tissue in chrysophanol dose group were(0.32±0.06)g,(0.24±0.07)g and(0.15±0.04)g,respectively(P<0.05 or P<0.01),and chry inhibited expressions of Ki67 and PCNA(P<0.05 or P<0.01),and regulated expression levels of AMPK,p-mTOR and cyclin D1 in tumor tissues in dose-dependent manner(P<0.05 or P<0.01).Conclusion:Chr inhibit proliferation,invasion and tumorigenesis of colon cancer SW480 cells by AMPK signaling pathway.
作者
代继源
刘文杰
王春辉
DAI Ji-Yuan;LIU Wen-Jie;WANG Chun-Hui(Department of Gastrointestinal and Anorectal Surgery,Shandong Changyi People′s Hospital,Changyi 261300,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2020年第14期1688-1694,共7页
Chinese Journal of Immunology
基金
山东省科技计划项目(2016YD14069)资助。
