巨噬细胞移动抑制因子与趋化因子受体7对结肠癌细胞侵袭及奥沙利铂耐药的影响  

Effect of MIF and CXCR7 on Invasion of Colon Cancer Cells and Oxaliplatin Resistance

作　　者：付伟杰 陈振海 杨钱 胡淞 FU Weijie;CHEN Zhenhai;YANG Qian;HU Song(Chongqing University Central Hospital·Chongqing Emergency Medical Center·The Fourth People′s Hospital of Chongqing·Chongqing Emergency Key Laboratory)

机构地区：[1]重庆大学附属中心医院·重庆市急救医疗中心·重庆市第四人民医院·重庆市急诊重点实验室,重庆400016

出　　处：《中国药业》2024年第21期48-52,共5页China Pharmaceuticals

基　　金：重庆市渝中区基础研究与前沿探索项目[20210177]。

摘　　要：目的探讨巨噬细胞移动抑制因子(MIF)与趋化因子受体7(CXCR7)对结肠癌细胞侵袭能力及奥沙利铂(L-OPH)耐药的影响。方法体外培养L-OPH结肠癌耐药细胞株HCT116/L-OPH和结肠癌敏感细胞株HCT116,设置对照组(A组,HCT116细胞株)和实验组[B组(HCT116/L-OPH细胞株),C组(shRNA转染技术沉默HCT116/L-OPH细胞株中CXCR7的表达,CXCR7-shRNA),D组(HCT116/L-OPH细胞株阴性对照,CXCR7-shCtrl),E组(HCT116/L-OPH加入MIF特异性小分子拮抗剂ISO-1),F组(CXCR7-shRNA加入ISO-1),G组(CXCR7-shCtrl加入ISO-1)],采用噻唑蓝(MTT)法检测各细胞组的耐药性及在不同L-OPH浓度下的细胞增殖活性,采用Transwell法检测细胞的侵袭能力,采用免疫印迹(Western blot)法检测CXCRT mRNA的表达水平和磷酸化丝氨酸-苏氨酸蛋白激酶(p-Akt)的表达水平,采用逆转录实时定量聚合酶链式反应(RT-qPCR)法检测CXCR7蛋白的表达水平。结果A组、B组、C组、D组细胞对L-OHP的半数抑制浓度分别为6.36,74.72,7.01,7.49μmol/L,B组细胞的耐药指数为11.74。10,20,50,100μmol/L L-OHP浓度下,B组细胞的增殖抑制率显著低于A组(P<0.05),C组细胞的增殖抑制率显著高于B组和D组(P<0.05),E组细胞的增殖抑制率显著高于B组(P<0.05)。C组细胞CXCR7的mRNA和蛋白表达水平均显著低于B组(P<0.05)。C组和E组细胞的侵袭能力显著弱于B组(P<0.05)。C组和E组细胞的p-Akt表达水平显著弱于B组(P<0.05)。结论MIF与CXCR7均可调控结肠癌细胞侵袭及L-OPH耐药,其作用机制可能与MIF通过CXCR7激活Akt信号通路相关。Objective To investigate the effect of macrophage migration inhibitory factor (MIF) and chemokine receptor 7 (CXCR7)on the invasive ability of colon cancer cells and their resistance to oxaliplatin (L-OPH).Methods Colon cancer cell line HCT116/L-OPH resistant to L-OPH and colon cancer sensitive cell line HCT116 in vitro were cultivated.The experiment set up the control group (group A,HCT116 cell line) and the experimental groups[group B (HCT116/L-OPPH cell line),group C (shRNA transfection technology silenced CXCR7 expression in HCT116/L-OPPH cell line,CXCR7-shRNA),group D (HCT116/L-OPPH cell line negative control,CXCR7-shCtrl),group E (HCT116/L-OPPH+MIF specific small molecule antagonist ISO-1),group F (CXCR7-shRNA+ISO-1),and group G (CXCR7-shCtrl+ISO-1)].MTT assay was used to detect the drug resistance and cell proliferation activity of each cell group at different L-OPPH concentrations.Transwell method was used to detect the invasive ability of cells.Western blot was used to detect the expression levels of CXCRT mRNA and phosphorylated serine-threonine (p-Akt).Reverse transcription real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the expression levels of CXCR7 protein.Results The half-maximal inhibitory concentrations (IC50) of cells against L-OHP in groups A,B,C,and D were 6.36,74.72,7.01,and 7.49μmol/L,respectively.The resistance index of cells in group B was 11.74.At concentrations of 10,20,50,and 100μmol/L of L-OHP,the cell proliferation inhibition rate in group B was significantly lower than that in group A (P<0.05),the proliferation inhibition rate of cells in group C was significantly higher than that in group B and group D (P<0.05),and the proliferation inhibition rate of cells in group E was significantly higher than that in group B (P<0.05).The expression levels of CXCR7 mRNA and protein in group C were significantly lower than those in group B (P<0.05).The invasion ability of cells in group E was significantly weaker than that in group B (P<0.05).The expression levels o

关 键 词：巨噬细胞移动抑制因子 趋化因子受体7 结肠癌 奥沙利铂 耐药 细胞侵袭 

分 类 号：R965[医药卫生—药理学] R979.1[医药卫生—药学]