微小RNA-6768-5p在肺癌组织中的表达及对肺癌细胞恶性生物学行为的影响  

Expression of microRNA-6768-5p in lung cancer tissue and its effect on malignant biological behavior of lung cancer cells

作　　者：毛万里 胡攀攀 邹纪忠 周耀东[3] 刘良文 MAO Wanli;HU Panpan;ZOU Jizhong;ZHOU Yaodong;LIU Liangwen(Department of Oncology,People′s Hospital of Yongchuan District,Chongqing 402160,China;Department of Clinical Laboratory,Center for Disease Control and Prevention,Yongchuan District,Chongqing 402160,China;Department of Oncology,Ruijin Hospital Affiliated to Shanghai Jiaotong University,Shanghai 200030,China)

机构地区：[1]重庆市永川区人民医院肿瘤科,重庆402160 [2]重庆市永川区疾病预防与控制中心检验科,重庆402160 [3]上海交通大学附属瑞金医院肿瘤科,上海200030

出　　处：《国际检验医学杂志》2024年第4期392-396,403,共6页International Journal of Laboratory Medicine

基　　金：国家自然科学基金项目(81902310)。

摘　　要：目的 探讨miR-6768-5p在肺癌组织中的表达及通过靶向调控羧肽酶A4(CPA4)对肺癌细胞增殖及侵袭的影响。方法 使用TCGA数据库分析肺癌组织和癌旁组织中miR-6768-5p的表达,使用实时定量PCR(qPCR)检测人肺癌细胞系(HCC1588、H1650、H1299、A549、HCC827)和正常肺泡上皮细胞(HPAEpiC细胞)中miR-6768-5p的表达。分别转染NC mimics、miR-6768-5p mimics至肺癌细胞,分别作为NC组和miR-6768-5p组。使用MTS实验、Matrigel侵袭实验分别检测各组细胞增殖和侵袭能力。使用RNAhybrid软件和双荧光素酶报告基因实验验证miR-6768-5p和CPA4的假定结合位点。qPCR检测各组细胞中CPA4 mRNA的表达。通过Western blot分析各组细胞中AKT/c-MYC信号通路蛋白的表达情况。结果 与癌旁组织比较,miR-6768-5p相对表达水平在肺癌组织中明显降低,差异有统计学意义(P<0.05)。与HPAEpiC细胞比较,miR-6768-5p相对表达水平在肺癌细胞系中明显降低,差异有统计学意义(P<0.05)。与NC组比较,miR-6768-5p组细胞增殖率明显降低(P<0.05)。NC组和miR-6768-5p组侵袭细胞数分别为(131.30±12.55)个和(37.45±7.77)个,miR-6768-5p组侵袭细胞数明显低于NC组(P<0.05)。miR-6768-5p组H1299细胞内CPA4 mRNA相对表达水平明显低于NC组(t=4.93,P<0.05)。与NC组比较,miR-6768-5p组AKT/c-MYC信号通路蛋白p-AKT、p-mTORC1、XIAP、MDM2、c-MYC蛋白表达明显降低。结论 肺癌组织中miR-6768-5p表达降低,miR-6768-5p可能通过靶向CPA4,抑制AKT/c-MYC信号通路激活,降低肺癌H1299细胞的增殖和侵袭能力。Objective To investigate the expression of miR-6768-5p in lung cancer tissue and its effect on the proliferation and invasion of lung cancer cells through targeted regulation of carboxypeptidase A4(CPA4).Methods The expression of miR-6768-5p in lung cancer tissues and adjacent tissues was analyzed using the TCGA database.Quantitative real-time PCR(qPCR)was used to detect the expression of miR-6768-5p in human lung cancer cell lines(HCC1588,H1650,H1299,A549,HCC827)and normal alveolar epithelial cells(HPAEpiC cells).Lung cancer cells were transfected with NC mimics and miR-6768-5p mimics,respectively,and divided into NC group and miR-6768-5p group.The MTS assay and Matrigel invasion assay were used to detect the cell proliferation and invasion ability of each group,respectively.The putative binding sites of miR-6768-5p and CPA4 were verified using RNAhybrid software and dual-luciferase reporter gene experiment.The expression of CPA4 mRNA in each group of cells was detected by qPCR.The expression of AKT/c-MYC signaling pathway proteins in the cells of each group was analyzed by Western blot.Results Compared with the adjacent tissues,the relative expression level of miR-6768-5p in lung cancer tissues was significantly decreased,and the difference was statistically significant(P<0.05).Compared with HPAEpiC cells,the relative expression level of miR-6768-5p was significantly decreased in lung cancer cell lines,and the difference was statistically significant(P<0.05).Compared with the NC group,the cell proliferation rate of miR-6768-5p group was significantly decreased(P<0.05).The number of invasive cells in NC group and miR-6768-5p group was(131.30±12.55)and(37.45±7.77),respectively,and the number of invasive cells in miR-6768-5p group was significantly lower than that in NC group(P<0.05).The relative expression level of CPA4 mRNA in H1299 cells of miR-6768-5p group was significantly lower than that in NC group(t=4.93,P<0.05).Compared with the NC group,the expressions of AKT/c-myC signaling pathway proteins p-AKT,p-m

关 键 词：miR-6768-5p 肺癌 羧肽酶A4 AKT/c-MYC信号通路 

分 类 号：R734.2[医药卫生—肿瘤]