RNAi沉默N-cadherin通过MEK-ERK信号通路抑制卵巢癌细胞增殖、侵袭、迁移及上皮-间质转化过程  

RNAi silencing N-cadherin inhibits the proliferation,invasion,migration and epithelial-mesenchymal transition of ovarian cancer cells through the MEK-ERK signaling pathway

作　　者：陈虹 潘琦文[1] 李建湘[1] 蓝燕[1] 潘彩萍[1] CHEN Hong;PAN Qiwen;LI Jianxiang;LAN Yan;PAN Caiping(Hechi People's Hospital,Hechi 547000,Guangxi,China)

机构地区：[1]河池市人民医院,广西河池547000

出　　处：《西部医学》2022年第8期1140-1146,共7页Medical Journal of West China

摘　　要：目的探讨沉默神经性钙黏附蛋白(N-cadherin)对卵巢癌细胞增殖、侵袭、迁移与上皮-间质转化的影响及分子机制。方法收集2021年3月~2021年12月我院卵巢癌患者手术切除的卵巢癌组织与邻近癌旁正常组织,采用免疫组织化学染色检测N-cadherin蛋白表达。通过RNAi技术沉默卵巢癌细胞SKOV3细胞中N-cadherin的表达后分为空白对照组、空载体组、siRNA-N-cadherin组。CCK-8法、细胞克隆形成实验、流式细胞术分别检测各组细胞增殖活性、克隆形成能力以及细胞周期分布,Transwell实验和细胞划痕实验分别检测各组细胞侵袭与迁移能力,实时荧光定量PCR检测各组细胞中N-cadherin、上皮性钙粘附分子(E-cadherin)mRNA表达,免疫细胞化学染色检测各组细胞中N-cadherin、E-cadherin、基质金属蛋白酶2(MMP2)和基质金属蛋白酶9(MMP9)蛋白表达,Western blot检测各组细胞中MEK、ERK蛋白表达。结果与癌旁正常组织比较,卵巢癌组织中N-cadherin呈高表达(P<0.05)。与空白对照组和空载体组比较,沉默N-cadherin能够抑制SKOV3细胞增殖活性(P<0.05),细胞克隆形成数目下降(P<0.01),并使细胞发生S期阻滞(P<0.01),细胞侵袭数目和迁移数目均减少(P<0.01);此外,沉默N-cadherin后SKOV3细胞中N-cadherin mRNA相对表达量下降,E-cadherin mRNA相对表达量升高(P<0.05),细胞内N-cadherin、MMP2和MMP9蛋白阳性染色减弱,E-cadherin蛋白阳性染色增强,细胞中MEK、ERK的蛋白表达水平均下降(P<0.05)。结论沉默N-cadherin可抑制卵巢癌细胞活性、细胞克隆形成、侵袭、迁移与上皮-间质转化的能力,使细胞阻滞于S期,其机制可能与阻碍MEK、ERK信号通路有关。Objective To explore the effect of silencing neuronal cadherin(N-cadherin)on the proliferation,invasion,migration and epithelial-mesenchymal transition of ovarian cancer cells and its molecular mechanism.Methods Immunohistochemical staining was used to detect the expression of N-cadherin protein in ovarian cancer tissues and adjacent normal tissues adjacent to the cancer.The expression of N-cadherin in SKOV3 cells was silenced by RNAi technology.CCK-8 method,cell clone formation experiment and flow cytometry respectively detected the proliferation activity,colony forming ability and cell cycle distribution of each group of cells.Transwell experiment and cell scratch experiment respectively detected the invasion and migration ability of each group of cells.The real-time fluorescent quantitative PCR detected the mRNA expression levels of N-cadherin and E-cadherin in each group of cells.The immunocytochemical staining detected the protein expression levels of N-cadherin,E-cadherin,matrix metalloproteinase 2(MMP2)and matrix metalloproteinase 9(MMP9)in each group of cells.Western blot detected the protein expression levels of MEK and ERK in each group of cells.Results Compared with normal tissues adjacent to cancer,N-cadherin was highly expressed in ovarian cancer tissues(P<0.05).Compared with the blank control group and the empty vector group,after SKOV3 cells silenced the expression of N-cadherin,the proliferation activity of SKOV3 cells was inhibited(P<0.05),the number of cell clone formation decreased(P<0.01),and the cells developed S Phase block(P<0.01),the number of cell invasion and migration were reduced(P<0.01).In addition,the relative mRNA expression of N-cadherin and E-cadherin in SKOV3 cells decreased after silencing N-cadherin,while the relative mRNA expression of E-cadherin increased(P<0.05),the positive staining of N-cadherin,MMP2 and MMP9 in the cells was weakened,the positive staining of E-cadherin increased,and the protein expression levels of MEK and ERK in the cells decreased(P<0.05).Conclusion Sil

关 键 词：N-CADHERIN 卵巢癌 增殖 侵袭 迁移 上皮-间质转化 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R737.31[医药卫生—基础医学]