CXCL1在乳癌MCF-7细胞上皮-间质转化中作用
EFFECT OF CHEMOKINE CXCL1 ON EPITHELIAL-MESENCHYMAL TRANSITION OF BREAST CANCER MCF-7 CELLS
马凯 1唐国庆 1张倩 2沈若武 3付兴芹 1张蓓1
作者信息
- 1. 青岛大学基础医学院免疫学系,山东青岛266071
- 2. 青岛阜外心血管病医院妇产科
- 3. 青岛大学基础医学院人体形态学实验室,山东青岛266071
- 折叠
摘要
目的 探讨趋化因子CXC型趋化因子配体1(CXCL1)对人乳癌细胞MCF-7迁移能力和上皮-间质转化(EMT)的影响及其机制.方法 体外培养人乳癌细胞MCF-7,分为对照组、CXCL1处理组、CXC趋化因子受体2(CXCR2)抑制剂组,应用CCK8方法检测各组细胞增殖活性,以ELISA法检测各组细胞E-cadherin、Vimentin、p-PI3K、p-AKT蛋白表达,RT-PCR方法检测E-cadherin、Vimentin mRNA表达.结果 CCK8检测结果显示,与对照组比较,CXCL1处理组细胞增殖活性升高(F=58.89,q=8.96,P＜0.01),CXCR2抑制剂组细胞增殖活性降低(q=6.31,P＜0.05).RT-PCR结果显示,与对照组相比较,CXCL1处理组E-cadherin mRNA表达下调(t=4.07,P＜0.01),Vimentin mRNA表达上调(t=3.62,P＜0.05).ELISA检测结果显示,与对照组比较,CXCL1处理组E-cadherin蛋白表达下调(F=8.95,q=2.02,P＜0.05),Vimentin蛋白表达上调(F=111.50,q=14.48,P＜0.01),CXCR2抑制剂组E-cadherin表达上调(q=2.04,P＜0.05),Vimentin蛋白表达下调(q=6.08,P＜0.05);与对照组比较,CXCL1处理组PI3K、PAKT蛋白表达明显升高(F=60.35、28.15,q=10.52、5.37,P＜0.01),CXCR2抑制剂组细胞内p-PI3K、p-AKT蛋白表达降低(q=4.65、4.97,P＜0.05).结论 乳癌细胞可能通过CXCL1/CXCR2轴活化PI3K/AKT信号通路发生EMT.
Abstract
Objective To investigate the effects of chemokine (CXC motif) ligand 1 (CXCL1) on the migration and epithelial-mesenchymal transition (EMT) of human breast cancer cells (MCF-7) and its mechanism.Methods MCF-7 cells were cultured in vitro and divided into three groups:control group,CXCL1 (40 μg/L) group,and CXC chemokine receptor 2 (CXCR2) inhibitor (SB225002 10 mmol/L) group.Cell Counting Kit-8 (CCK8) was used to evaluate the proliferative activity of MCF-7 cells in all groups.Furthermore,enzyme-linked immunosorbent assay (ELISA) was used to measure the protein expression of E-cadherin,vimentin,p-PI3K,and p-AKT.The expression of E-cadherin mRNA and Vimentin mRNA was measured by RT-PCR.Results CCK8 assay results showed that compared with the control group,the CXCL1 group had a significantly higher cell proliferative activity (F=58.89,q =8.96,P ＜0.01),while the CXCR2 inhibitor group had a significantly lower cell proliferative activity (q =6.31,P＜0.05).The RT-PCR results indicated that the CXCL1 group had significantly lower expression of E-cadherin mRNA and significantly higher expression of Vimentin mRNA than the control group (t =4.07,3.62;P ＜ 0.05).The ELISA results showed that the CXCL1 group had significantly lower protein expression of E-cadherin and significantly higher protein expression of Vimentin (F =8.95,q =2.02,P ＜0.05;F =111.50,q =14.48,P ＜0.01);while the CXCR2 inhibitor group had significantly higher protein expression of E-cadherin and significantly lower protein expression of Vimentin (q =2.04,6.08;P ＜ 0.05).The CXCL1 group had significantly higher protein expression of p-PI3K and p-AKT than the control group (F =60.35,q =10.52,P＜0.01;F =28.15,q =5.37,P＜0.01);the CXCR2 inhibitor group had significantly lower protein expression of p-PI3K and p-AKT than the control group (q=4.65,4.97;P＜0.05).Conclusion Chemokine CXCL1 plays a key role in EMT of MCF-7 cells by activating the PI3K/AKT signaling pathway through CXCL1/CXCR2 axis.
关键词
CXC型趋化因子配体1/CXC趋化因子受体2/乳房肿瘤/上皮-间质转化/迁移Key words
CXC chemokine ligand 1/CXC chemokine receptor 2/breast neoplasms/epithelial-mesenchymal transition/migration引用本文复制引用
基金项目
山东省高等学校科技计划项目(J14LK10)
山东省高等学校科技计划项目(J16LK-04)
出版年
2017
NETL
NSTL
维普
万方数据