circ_0026134调控miR-1270/GRB2途径影响肝癌细胞的放射敏感性  

Circ_0026134 regulates the miR-1270/GRB2 pathway to affect the radiosensitivity of hepatoma cells

作　　者：赵晓阳 刘玉岩 李陆鹏 刘建文 Zhao Xiaoyang;Liu Yuyan;Li Lupeng;Liu Jianwen(Department of Comprehensive Intervention,Henan Provincial People's Hospital People's Hospital of Zhengzhou University Zhengzhou 450000,China)

机构地区：[1]河南省人民医院、郑州大学人民医院综合介入科,郑州450000

出　　处：《中华肝脏病杂志》2024年第1期40-48,共9页Chinese Journal of Hepatology

摘　　要：目的探讨环状RNA 0026134(circ_0026134)是否通过调控miR-1270/生长因子受体结合蛋白2(GRB2)途径影响肝癌细胞的放射敏感性。方法实时定量PCR(RT-qPCR)检测肝癌组织、肝癌细胞中circ_0026134、miR-1270和GRB2表达量。生物信息学分析、双荧光素酶报告基因实验、RT-qPCR、免疫印迹法分析circ_0026134与miR-1270、miR-1270与GRB2的靶向关系。细胞计数试剂盒、Transwell实验、划痕实验、流式细胞术检测circ_0026134、miR-1270、GRB2表达联合6 Gy对Huh7和SK-HEP-1细胞增殖、侵袭、迁移、凋亡的影响。裸鼠成瘤实验检测沉默circ_0026134对肿瘤形成的影响。计量资料以均数±标准差表示。两组间比较使用独立样本t检验,多组间比较采用单因素方差分析和SNK-q检验。P<0.05为差异有统计学意义。结果肝癌组织、Huh7和SK-HEP-1细胞中circ_0026134和GRB2表达量显著增加,而miR-1270表达量显著降低(P<0.05)。放疗后Huh7、SK-HEP-1中circ_0026134表达量显著降低(P<0.05)。circ_0026134与miR-1270直接结合并负调控miR-1270表达(P<0.05)。miR-1270与GRB2直接结合并负调控GRB2表达(P<0.05)。6 Gy辐射显著抑制Huh7和SK-HEP-1细胞增殖、迁移和侵袭,并诱导细胞凋亡(P<0.05)。沉默circ_0026134或过表达miR-1270显著增强6 Gy处理对肝癌细胞抗增殖、抗迁移侵袭和促凋亡作用(P<0.05)。抑制miR-1270显著减弱沉默circ_0026134联合6 Gy辐射对肝癌细胞增殖、迁移侵袭、凋亡的影响(P<0.05)。过表达GRB2显著减弱miR-1270过表达联合6 Gy辐射对肝癌细胞增殖、迁移侵袭、凋亡的影响(P<0.05)。敲减circ_0026134显著延缓体内肿瘤生长(P<0.05)。结论沉默circ_0026134通过负调控miR-1270/GRB2途径增强辐射处理对肝癌细胞的抗增殖、抗迁移侵袭以及促凋亡作用,增强肝癌细胞放射敏感性。Objective To investigate whether circular RNA 0026134(circ_0026134)affects the radiosensitivity of hepatoma cells by regulating the miR-1270/growth factor receptor-bound protein 2(GRB2)pathway.Methods Real-time quantitative PCR(RT-qPCR)was used to detect the expression levels of circ_0026134,miR-1270,and GRB2 in liver cancer tissues and cells.Bioinformatics analysis,a dual-luciferase gene reporter assay,RT-qPCR,and western blot were used to analyze the targeting relationships between circ_0026134 and miR-1270 and miR-1270 and GRB2.The effects of circ_0026134,miR-1270,and GRB2 expression combined with 6 Gy on the proliferation,invasion,migration,and apoptosis of Huh7 and SK-HEP-1 cells were detected by a cell counting kit,a transwell assay,a scratch assay,and flow cytometry.The tumorigenesis experiment was used to detect the effect of silencing circ_0026134 in nude mice.Measurement data are expressed as the mean±standard deviation.The independent sample t-test was used for comparison between two groups,and the one-way analysis of variance and SNK-q test were used for comparison between multiple groups.P<0.05 was considered statistically significant.Results The expression levels of circ_0026134 and GRB2,Huh7,and SK-HEP-1 cells in liver cancer tissues were significantly increased,while the expression levels of miR-1270 were significantly decreased(P<0.05).The expression of circ_0026134 in Huh7 and SK-HEP-1 decreased significantly after radiotherapy(P<0.05).circ_0026134 binds directly to miR-1270 and negatively regulates miR-1270 expression(P<0.05).miR-1270 binds directly to GRB2 and negatively regulates GRB2 expression(P<0.05).6 Gy radiation significantly inhibited the proliferation,migration,and invasion of Huh7 and SK-HEP-1 cells and induced apoptosis(P<0.05).Silencing circ_0026134 or overexpression of miR-1270 significantly enhanced the anti-proliferation,anti-migration,invasion,and pro-apoptosis effects of 6 Gy treatment on hepatoma cells(P<0.05).Inhibition of miR-1270 significantly weakened the effects of sile

关 键 词：肝癌 细胞增殖 凋亡 迁移侵袭 放射敏感性 环状RNA 0026134 微小RNA 1270 生长因子受体结合蛋白2 

分 类 号：R735.7[医药卫生—肿瘤]