LINC01915介导miR-92a影响结直肠癌裸鼠肿瘤生长的实验研究  

Experimental study on LINC01915 mediates miR-92a to affect tumor growth in nude mice with colorectal cancer

作　　者：韩炜[1] 李程[1] 李文翰 霍斌亮[1] 师文 HAN Wei;LI Cheng;LI Wenhan;HUO Binliang;SHI Wen(Department of Oncological Surgery,Shaanxi Provincial People′s Hospital,Xi′an,Shaanxi 710068,China;Department of Gastroenterology,First Affiliated Hospital of Xi′an Jiaotong University,Xi′an,Shaanxi 710061,China)

机构地区：[1]陕西省人民医院肿瘤外科,陕西西安710068 [2]西安交通大学第一附属医院消化内科,陕西西安710061

出　　处：《检验医学与临床》2025年第2期157-165,共9页Laboratory Medicine and Clinic

基　　金：陕西省自然科学基础研究计划项目(2021JQ-917);陕西省人民医院科技人才支持计划项目(2021JY-46)。

摘　　要：目的探讨基因间区长链非编码核糖核酸01915(LINC01915)过表达靶向抑制微小核糖核酸-92a(miR-92a)控制结直肠癌裸鼠肿瘤生长的作用与分子机制。方法取100只5周龄裸鼠经皮下接种人结直肠癌细胞HT29建立结直肠癌裸鼠皮下移植瘤模型,将建模成功的裸鼠随机分为LINC01915上调组(转染pcDNA-LINC01915)、LINC01915下调组(转染si-LINC01915)、LINC01915上调对照组(转染pcDNA)、LINC01915下调对照组(转染si-NC)、miR-92a上调组(转染miR-92a mimic)、miR-92a下调组(转染miR-92a inhibitor)、miR-92a上调对照组(转染miR-92a mimic NC)、miR-92a下调对照组(转染miR-92a inhibitor NC)以及空白对照组(注射生理盐水)。观察记录裸鼠肿瘤体积并绘制肿瘤生长曲线;通过苏木精-伊红(HE)染色观察肿瘤组织病理学改变;采用实时荧光定量聚合酶链反应(RT-qPCR)及蛋白质印迹法(WB)检测裸鼠肿瘤组织Kruppel样因子4(KLF4)、生存素(Survivin)、细胞增殖核抗原(Ki-67)及半胱氨酸天冬氨酸特异性蛋白酶-3(Caspase-3)信使核糖核酸(mRNA)或蛋白表达;通过荧光素酶活性实验验证LINC01915和miR-92a的靶向关系。结果重复测量方差分析结果显示,各组结直肠癌裸鼠肿瘤体积变化存在时间效应、组间效应及交互效应,差异均有统计学意义(P<0.05)。多变量方差分析结果显示,转染第9、12、15天时:与空白对照组和LINC01915上调对照组相比,LINC01915上调组结直肠癌裸鼠肿瘤体积缩小(P<0.05);与空白对照组和LINC01915下调对照组相比,LINC01915下调组结直肠癌裸鼠肿瘤体积均增大(P<0.05);与空白对照组和miR-92a上调对照组相比,miR-92a上调组结直肠癌裸鼠肿瘤体积均增大(P<0.05);与空白对照组和miR-92a下调对照组相比,miR-92a下调组结直肠癌裸鼠肿瘤体积均缩小(P<0.05)。各对照组裸鼠肿瘤组织腺体排列紊乱并伴随炎症细胞浸润,有少量杯状细胞;LINC01915上调组和miR-92a下调组裸鼠肿瘤组织腺�Objective To investigate the effect and molecular mechanism of long non-coding RNA 01915(LINC01915)overexpression on controlling on tumor growth in nude mice with colorectal cancer by targeted inhibition of microRNA-92a(miR-92a).Methods A total of 1005-week-old nude mice were subcutaneously inoculated with human colorectal cancer cells HT29 to establish a subcutaneously transplanted tumor model of colorectal cancer in nude mice.The nude mice with successful model construction were randomly divided into the LINC01915 up-regulation group(transfected pcDNA-LINC01915),LINC01915 down-regulation group(transfected si-LINC01915),LINC01915 up-regulation control group(transfected pcDNA),LINC01915 down-regulation control group(transfected si-NC),miR-92a up-regulated group(transfected miR-92a mimic),miR-92a down-regulation group(transfected miR-92a inhibitor),miR-92a up-regulated control group(transfection miR-92a mimic NC),miR-92a down-regulation control group(transfected miR-92a inhibitor NC)and blank control group(injection of normal saline).The tumor volume of nude mice was observed and recorded,and the tumor growth curve was drawn.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of the tumor tissues.The real-time quantitative polymerase chain reaction(RT-qPCR)and Western blotting(WB)were used to detect the Kruppel like factor 4(KLF4),Survivin,cell proliferating nuclear antigen(Ki-67)and cysteine aspartic protease-3(Caspase-3)mRNA or protein expression in nude mice tumor tissues;the luciferase activity experiment was used to verify the targeting relationship between LINC01915 and miR-92a.Results The repeated measures ANOVA results showed that the time effect,inter-group effect and interaction effect of colorectal cancer nude mouse tumor volumes in each group were all statistically significant(P<0.05).The multivariate ANOVA analysis revealed that on post-transfection 9,12 and 15 d:compared with the blank control group and LINC01915 up-regulation control group,the tumor volume of nude mice wi

关 键 词：基因间区长链非编码核糖核酸 结直肠癌 miR-92a Kruppel样因子4 半胱氨酸天冬氨酸特异性蛋白酶-3 

分 类 号：R574.63[医药卫生—消化系统] R446.1[医药卫生—内科学]