环状RNA HN1靶向miR-766-5p/KLF3轴调控胃癌细胞生物学行为的分子机制  

Molecular mechanism of circular RNA HN1 in regulating the biological behavior of gastric cancer cells by targeting the miR⁃766⁃5p/KLF3 Axis

作　　者：王莉[1] 师红利 李峰 刘泓[1] WANG Li;SHI Hong-li;LI Feng;LIU Hong(Department of Gastroenterology,Linfen Peoples Hospital,Linfen,Shanxi Province 041000,China)

机构地区：[1]临汾市人民医院消化内科,山西临汾041000

出　　处：《解剖学研究》2025年第1期18-24,31,共8页Anatomy Research

基　　金：山西省教育厅高校科技创新计划各类项目(2020L0225)。

摘　　要：目的探讨环状RNA HN1(CircHN1)靶向微小RNA-766-5p(miR-766-5p)/Krüpple样转录因子3(KLF3)轴调控胃癌细胞生物学行为的分子机制。方法实时荧光定量PCR(qRT-PCR)、western blot分别检测人胃癌细胞系MKN-45以及人正常胃黏膜上皮细胞GES-1中CircHN1、miR-766-5p、KLF3表达;将si-NC、si-CircHN1、si-CircHN1+inhibitor NC、si-CircHN1+miR-766-5p inhibitor分别转染至MKN-45细胞并命名为si-NC组、si-CircHN1组、si-CircHN1+inhibitor NC组、si-CircHN1+miR-766-5p inhibitor组,未做任何处理的MKN-45细胞记为NC组。双荧光素酶报告基因实验检测CircHN1、miR-766-5p、KLF3的关系;qRT-PCR检测MKN-45细胞中CircHN1、miR-766-5p表达;Western blot检测MKN-45细胞KLF3、上皮钙黏附素(E-cadherin)、神经型钙黏附蛋白(N-cadherin)、波形蛋白(Vimentin)表达;MTT法检测MKN-45细胞活性;流式细胞术检测MKN-45细胞凋亡率;Transwell检测MKN-45细胞侵袭、迁移情况。结果与GES-1细胞(1.00±0.00)相比,MKN-45细胞中CircHN1水平(1.48±0.16)、KLF3水平(1.17±0.13)显著上调(P<0.05),miR-766-5p表达量(0.32±0.05)显著下调(P<0.05);与NC组、si-NC组相比,si-CircHN1组MKN-45细胞CircHN1表达量、KLF3、N-cadherin、Vimentin蛋白水平、A570值(0.35±0.03)、迁移、侵袭细胞数量显著降低(P<0.05),MKN-45细胞凋亡率(25.57%±3.26%)、miR-766-5p表达量、E-cadherin蛋白水平显著升高(P<0.05);而miR-766-5p下调减弱了沉默CircHN1抑制MKN-45细胞增殖、迁移和侵袭的有利影响;CircHN1靶向调节miR-766-5p/KLF3轴。结论沉默CircHN1可能通过上调miR-766-5p来抑制KLF3表达,进而抑制MKN-45细胞恶性生物学行为。Objective To investigate the molecular mechanism of circular RNA HN1(CircHN1)in regulat⁃ing the biological behavior of gastric cancer cells by targeting the micro RNA 766⁃5p(miR⁃766⁃5p)/Krüpple like transcription factor 3(KLF3)axis.Methods Real⁃time fluorescent quantitative PCR(qRT⁃PCR)and western blot were applied to detect the expression of CircHN1,miR⁃766⁃5p and KLF3 in human gastric cancer cell line MKN⁃45 and human normal gastric mucosa epithelial cell line GES⁃1,respectively;MKN⁃45 cells were transfected with si⁃NC,si⁃CircHN1,si⁃CircHN1+inhibitor NC,si⁃CircHN1+miR⁃766⁃5p inhibitor and named as si⁃NC group,si⁃CircHN1 group,si⁃CircHN1+inhibitor NC group,and si⁃CircHN1+miR⁃766⁃5p inhibitor group.MKN⁃45 cells with⁃out any treatment were recorded as NC group.Double luciferase reporter gene experiment was applied to detect the re⁃lationship between CircHN1,miR⁃766⁃5p and KLF3;qRT⁃PCR was applied to detect the expression of CircHN1 and miR⁃766⁃5p in MKN⁃45 cells;Western blot was applied to detect the expression of KLF3,E⁃cadherin,N⁃cad⁃herin and Vimentin in MKN⁃45 cells;MTT method was applied to detect the activity of MKN⁃45 cells;flow cytome⁃try was applied to detect the apoptosis rate of MKN⁃45 cells;Transwell was applied to detect the invasion and migra⁃tion of MKN⁃45 cells.Results Compared with GES⁃1 cells,the levels of CircHN1(1.48±0.16)and KLF3(1.17±0.13)in MKN⁃45 cells were obviously up⁃regulated(P<0.05),the expression level of miR⁃766⁃5p(0.32±0.05)was obviously down⁃regulated(P<0.05).Compared with the NC group and si⁃NC group,the expression level of CircHN1,and the protein levels of KLF3,N⁃cadherin,Vimentin in MKN⁃45 cells,A570 value(0.35±0.03),and the numbers of migration,and invasion cells in the si⁃CircHN1 group were obviously reduced(P<0.05),the apoptosis rate(25.57%±3.26%),the expression level of miR⁃766⁃5p,and the protein level of E⁃cadherin in MKN⁃45 cells were obviously increased(P<0.

关 键 词：胃癌 环状RNA HN1 微小RNA-766-5p Krüpple样转录因子3 增殖 迁移 侵袭 

分 类 号：R735.2[医药卫生—肿瘤]