肿瘤蛋白翻译控制反义RNA1靶向调控miR-30c-5p对肝癌细胞放射敏感性和增殖迁移及侵袭的影响  

Effects of tumor protein translation control antisense RNA1 on radiosensitivity,proliferation,migration and invasion of hepatocellular carcinoma cells by targeting miR-30c-5p

作　　者：翟金俊[1] 杜贤荣[2] 李彩霞[1] Zhai Jinjun;Du Xianrong;Li Caixia(Department of Emergency,Shanxi People′s Hospital,Taiyuan 030000,China;Department of Oncology,Shanxi People′s Hospital,Taiyuan 030000,China)

机构地区：[1]山西省人民医院急诊科,太原030000 [2]山西省人民医院肿瘤科,太原030000

出　　处：《中华肿瘤杂志》2021年第10期1054-1061,共8页Chinese Journal of Oncology

摘　　要：目的探讨肿瘤蛋白翻译控制反义RNA1(TPT1-AS1)靶向miR-30c-5p对肝癌细胞放射敏感性、细胞增殖、迁移和侵袭的影响。方法34例肝癌组织及癌旁正常组织来源于2016年3月至2018年3月就诊于山西省人民医院的肝癌患者。将肝癌HepG2细胞分为si-NC组(转染si-NC)、si-TPT1-AS1组(转染si-TPT1-AS1)、pcDNA3.1组(转染pcDNA3.1)、pcDNA3.1-TPT1-AS1组(转染pcDNA3.1-TPT1-AS1)、si-TPT1-AS1+anti-miR-NC组(转染si-TPT1-AS1和anti-miR-NC)、si-TPT1-AS1+anti-miR-30c-5p组(转染si-TPT1-AS1+anti-miR-30c-5p)。采用实时荧光定量聚合酶链反应检测癌旁正常组织和肝癌组织中TPT1-AS1和miR-30c-5p的基因表达,克隆形成实验检测细胞放射敏感性,MTT实验检测细胞增殖活力,流式细胞数检测细胞周期分布,Transwell实验检测细胞迁移和侵袭能力,双荧光素酶报告基因实验验证TPT1-AS1和miR-30c-5p的靶向关系,Western blot检测增殖、迁移和侵袭相关蛋白的表达。结果肝癌组织中TPT1-AS1和miR-30c-5p的表达水平为0.84±0.08和0.13±0.01,与癌旁正常组织(分别为0.31±0.03和0.50±0.05)比较,差异均有统计学意义(均P<0.05)。采用2、4、6、8 Gy照射细胞时,si-TPT1-AS1组细胞存活分数分别为0.280±0.040、0.069±0.011、0.020±0.003和0.005±0.001,均低于si-NC组(分别为0.648±0.070、0.348±0.080、0.130±0.020和0.060±0.009,均P<0.05),si-TPT1-AS1组细胞放射增敏比为1.672。si-TPT1-AS1组细胞迁移数、侵袭数分别为(50.00±4.36)个和(44.00±4.03)个,低于si-NC组[分别为(109.00±8.68)个和(94.00±7.49)个,均P<0.05]。培养24、48和72 h,si-TPT1-AS1组细胞吸光度(A)值分别为0.28±0.03、0.43±0.04和0.68±0.07,低于si-NC组(分别为0.46±0.04、0.87±0.08和1.35±0.13,均P<0.05)。si-TPT1-AS1组细胞周期素D1(Cyclin D1)、p21、E-cadherin和基质金属蛋白酶2(MMP-2)蛋白表达水平分别为0.25±0.02、0.65±0.06、0.68±0.07和0.27±0.03,与si-NC组(分别为0.88±0.08、0.17±0.02、0.14±0.01和0.89±0.09)比较Objective To investigate the effects of tumor protein translation control antisense RNA1(TPT1-AS1)on the radiosensitivity,cell proliferation,migration and invasion of hepatocellular carcinoma cells by targeting microRNA-30c-5p(miR-30c-5p).Methods Thirty-four cases of liver cancer tissues and adjacent normal tissues were derived from liver cancer patients who were admitted to Shanxi Provincial People′s Hospital from March 2016 to March 2018.Liver cancer HepG2 cell was transfected with negative control siRNA(si-NC group),si-TPT1-AS1(si-TPT1-AS1 group),pcDNA3.1(pcDNA3.1 group),pcDNA3.1-TPT1-AS1(pcDNA3.1-TPT1-AS1 group),si-TPT1-AS1 and anti-miR-NC(si-TPT1-AS1+anti-miR-NC group),si-TPT1-AS1 and anti-miR-30c-5p(si-TPT1-AS1+anti-miR-30c-5p group),respectively.Real-time quantitative reverse transcription polymerase chain reaction(qPCR)was used to detect the transcription levels of TPT1-AS1 and miR-30c-5p in normal tissues adjacent to cancer and liver cancer tissues,the clone formation test was used to test the radiosensitivity of HepG2 cells,and the Methyl Thiazolyl Tetrazolium(MTT)test was used to test the proliferation of HepG2 cells.Cell cycle distribution was detected by flow cytometry,Transwell array was used to detect the migration and invasion ability of HepG2 cells,dual luciferase reporter array was used to verify the targeting relationship of TPT1-AS1 and miR-30c-5p,western blot was used to detect the expressions of proliferation,migration and invasion-related proteins.Results The expression levels of TPT1-AS1 and miR-30c-5p in liver cancer tissues were 0.84±0.08 and 0.13±0.01,statistically different from 0.31±0.03 and 0.50±0.05 in normal tissues adjacent to cancer(P<0.05).When the cells were treated with 2,4,6,8 Gy irradiation,the cell survival scores of the si-TPT1-AS1 group were 0.280±0.040,0.069±0.011,0.020±0.003 and 0.005±0.001,respectively,lower than 0.648±0.070,0.348±0.080,0.130±0.020 and 0.060±0.009 of the si-NC group(P<0.05),the radiosensitization ratio of the si-TPT1-AS1 group was 1.672.T

关 键 词：肝肿瘤 肿瘤蛋白翻译控制反义RNA1 miR-30c-5p 放射敏感性 细胞增殖 迁移 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]