miR-30a-5p调控ECM-受体作用信号通路相关蛋白抑制肺腺癌细胞的侵袭和迁移  

miR-30a-5p inhibits invasion and migration of lung adenocarcinoma cells via regulating ECM-receptor signaling pathway

作　　者：夏建洪 周立庆 严研 马婷婷 苏欣宇 XIA Jianhong;ZHOU Liqing;YAN Yan;MA Tingting;SU Xinyu(Department of Radiotherapy,the Affiliated Huai'an Hospital of Xuzhou Medical University(The Second People's Hospital of Huai'an),Huai'an 223002,China;不详)

机构地区：[1]徐州医科大学附属淮安医院(淮安市第二人民医院)放疗科,223002

出　　处：《浙江医学》2023年第1期21-27,共7页Zhejiang Medical Journal

摘　　要：目的探讨miR-30a-5p和细胞外基质(ECM)-受体作用信号通路相关蛋白在肺腺癌细胞进展中发挥的调控作用。方法将miR-30a-5p模拟物及其阴性对照分别转染到肺腺癌Calu-3细胞内,设为miR-30a-5p模拟物组、模拟物对照组。通过基因集富集分析(GSEA)软件对miR-30a-5p进行通路富集分析。将miR-30a-5p抑制剂及其阴性对照转染到细胞内,设为miR-30a-5p抑制剂组、抑制剂对照组,在miR-30a-5p抑制剂组的细胞中加入ECM-受体相互作用通路蛋白抑制剂,设为miR-30a-5p抑制剂+ECM-受体相互作用抑制剂组。采用qRT-PCR法检测不同处理组细胞中miR-30a-5p mRNA表达水平,采用Western blot法检测肺腺癌细胞系中ECM-受体作用信号通路相关蛋白磷酸化水平,采用噻唑蓝(MTT)实验、细胞划痕愈合实验和Transwell实验分别检测各处理组细胞的增殖、迁移与侵袭能力。结果与人正常支气管上皮细胞BEAS-2B比较,肺腺癌细胞系SPC-A1、Calu-3、HCC827、PC14中miR-30a-5p表达水平下调(均P<0.05)。与模拟物对照组相比,miR-30a-5p模拟物抑制了细胞的增殖、迁移与侵袭(均P<0.05)。在Calu-3细胞中,miR-30a-5p显著富集在ECM-受体作用信号通路上。与抑制剂对照组相比,miR-30a-5p抑制剂组中ECM-受体作用信号通路相关蛋白磷酸化水平上调(均P<0.05),而miR-30a-5p抑制剂+ECM-受体相互作用抑制剂组相关蛋白磷酸化水平得到恢复。与抑制剂对照组相比,miR-30a-5p抑制剂组Calu-3细胞的增殖、迁移与侵袭上调(均P<0.05),miR-30a-5p抑制剂+ECM-受体相互作用抑制剂组Calu-3细胞的增殖、迁移与侵袭能力得到恢复。结论miR-30a-5p可能在肺腺癌细胞的发生、发展中发挥重要的作用,部分作用是通过调控ECM-受体作用信号通路相关蛋白实现的,miR-30a-5p可能是治疗肺腺癌的新靶点。Objective To explore the regulatory role of miR-30a-5p and extracellular matrix(ECM)-receptor signaling pathway related proteins in the progression of lung adenocarcinoma cells.Methods The miR-30a-5p mimic or its negative control was transfected into lung adenocarcinoma Calu-3 cells respectively,named miR-30a-5p mimic group and mimic control group.The pathway enrichment analysis of miR-30a-5p was analyzed by gene set enrichment analysis(GSEA)software.The miR-30a-5p inhibitor or its negative control were transfected into Calu-3 cells,respectively,named miR-30a-5p inhibitor group and inhibitor control group.ECM-receptor interaction pathway protein inhibitor was added in miR-30a-5p inhibitor group(miR-30a-5p inhibitor+ECM-receptor interaction inhibitor group).The mRNA expression level of miR-30a-5p in Calu-3 cells was detected by qRT-PCR.The phosphorylation of ECM-receptor signaling pathway related proteins in Calu-3 cells was determined by Western blot assay.MTT assay,cell scratch healing assay and Transwell assay were used to detect the proliferation,migration and invasion abilities of cells in each treatment group.Results Compared with human normal bronchial epithelial BEAS-2B cells,the expression of miR-30a-5p in lung adenocarcinoma cell lines SPC-A1,Calu-3,HCC827 and PC14 were significantly downregulated(all P<0.05).Compared with the mimic control group,miR-30a-5p mimic treatment significantly inhibited cell proliferation,migration and invasion(all P<0.05).In Calu-3 cells,miR-30a-5p was significantly enriched in the ECM-receptor signaling pathway.Compared with the inhibitor control group,the phosphorylation levels of ECM-receptor action signal pathway related proteins were significantly increased in the miR-30a-5p inhibitor group(all P<0.05),while in the miR-30a-5p inhibitor+ECM-receptor interaction inhibitor group,the phosphorylation levels of related proteins were restored.Compared with the inhibitor control group,the proliferation,migration and invasion of Calu-3 cells in miR-30a-5p inhibitor group were sign

关 键 词：miR-30a-5p 细胞外基质-受体作用信号通路 增殖 侵袭 迁移 

分 类 号：R734.2[医药卫生—肿瘤]