LPS对乳腺癌MDA-MB-231细胞上皮间质转化的诱导作用及其对β-catenin表达的影响  

Induction of LPS on epithelial mesenchymal transition in breast cancer MDA-MB-231 cells and its effect on β-catenin expression

作　　者：陈州华 龚辉 冯磊 肖玉洁[3] 黄立中[3] CHEN Zhouhua;GONG Hui;FENG Lei;XIAO Yujie;HUANG Lizhong(Department of Oncology,Second People’s Hospital of Xiangtan City,Hunan Province,Xiangtan 411100,China;Department of Oncology,Affiliated Hospital,Hunan Provincial Institute of Tranditional Chinese Medicine,Changsha 410006,China;Department of Internal Medicine,College of Integrated Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南省湘潭市第二人民医院肿瘤科,湖南湘潭411100 [2]湖南省中医药研究院附属医院肿瘤科,湖南长沙410006 [3]湖南中医药大学中西医结合学院内科教研室,湖南长沙410208

出　　处：《吉林大学学报（医学版）》2020年第2期309-315,共7页Journal of Jilin University：Medicine Edition

基　　金：湖南省教育厅重点项目资助课题(15A139);湖南省教育厅一般项目资助课题(17C1205)。

摘　　要：目的:探讨脂多糖(LPS)对人乳腺癌MDA-MB-231细胞中上皮间质转化(EMT)标志物及β-连环素(β-catenin)表达的影响,阐明其可能机制。方法:人乳腺癌MDA-MB-231细胞分为对照组和不同浓度(5、10、20和40 mg·L^-1)LPS组,倒置显微镜观察各组MDA-MB-231细胞的形态表现,免疫荧光实验检测各组MDA-MB-231细胞中β-catenin表达及定位,实时荧光定量PCR(RT-qPCR)法和Western blotting法检测各组MDA-MB-231细胞中EMT标志物E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)和β-catenin mRNA及蛋白表达水平。结果:对照组MDA-MB-231细胞形态表现为上皮细胞表型,不同浓度LPS组MDA-MB-231细胞形态表现为间质细胞表型。免疫荧光实验,β-catenin表达定位主要在细胞核。与对照组比较,不同浓度LPS组细胞中Vimentin和β-catenin mRNA及蛋白表达水平明显升高(P<0.05或P<0.01),以20 mg·L^-1 LPS组升高最为明显;与对照组比较,不同浓度LPS组细胞中E-cadherin mRNA和蛋白表达水平明显降低(P<0.05或P<0.01),以20 mg·L^-1 LPS组降低最为明显。结论:LPS通过下调E-cadherin表达、上调Vimentin表达促进乳腺癌MDA-MB-231细胞发生EMT、侵袭和转移,其作用机制可能与Wnt/β-catenin信号通路有关。Objective:To investigate the effect of lipopolysaccharide(LPS)on the expressions of epithelial-mesenchymal transition(EMT)markers andβ-catenin in the breast cancer MDA-MB-231 cells,and to clarify its possible mechanism.Methods:The breast cancer MDA-MB-231 cells were divided into control group and different concentrations(5,10,20,and 40 mg·L^-1)of LPS groups.Inverted microscope was used to observe the morphology of MDA-MB-231 cells in various groups.Immunofluorescence test was used to detect theβ-catenin expression and location in the MDA-MB-231 cells in various groups.Real-time quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the expression levels of the EMT markers E-cadherin,Vimentin andβ-catenin mRNA and proteins in the MDA-MB-231 cells in various groups.Results:The morphology of MDA-MB-231 cells in control group was epithelial phenotype,and the morphology of MDA-MB-231 cells in different concentrations of LPS groups were the phenotype of mesenchymal cells.The results of immunofluorescence staining showed that the expression ofβ-catenin was mainly located in the nucleus.Compared with control group,the expression levels of Vimentin andβ-catenin mRNA and proteins in the MDA-MB-231 cells in different concentrations of LPS groups were increased(P<0.05 or P<0.01),especially in 20 mg·L^-1 LPS group.Compared with control group,the expression levels of E-cadherin mRNA and proteins in the MDA-MB-231 cells in different concentrations of LPS groups were decreased(P<0.05 or P<0.01),especially in 20 mg·L^-1 LPS group.Conclusion:LPS could promote the EMT,invasion and metastasis of the breast cancer MDA-MB-231 cells by down-regulating the E-cadherin expression and up-regulating the Vimentin expression,and its mechanism may be related to Wnt/β-catenin signaling pathway.

关 键 词：乳腺肿瘤 脂多糖 上皮间质转化 WNT/Β-CATENIN信号通路 

分 类 号：R737.9[医药卫生—肿瘤]