LINC02532靶向miR-145影响胰腺癌干细胞的增殖、迁移、侵袭和凋亡  

Effects of LINC02532 targeting miR-145 on proliferation,migration,invasion and apoptosis of pancreatic cancer stem cells

作　　者：孟德峰 李长仔 吴春涛 Meng Defeng;Li Changzai;Wu Chuntao(Department of Oncology Surgery,North China University of Science and Technology Affiliated Hospital,Tangshan 063000,Hebei Province,China)

机构地区：[1]华北理工大学附属医院肿瘤外科,河北省唐山市063000

出　　处：《中国组织工程研究》2022年第1期52-58,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:长链非编码RNA(lncRNA)和微小RNA(miRNA)是两类参与调控细胞增殖、凋亡和迁移等生命活动的小分子非编码RNA,且lncRNA还可作为竞争性内源性RNA与miRNA靶向结合,调控miRNA靶基因的表达,在肿瘤的发生发展中起重要作用。LINC02532可靶向结合miR-129-5p和miR-490-5p促进胃癌细胞增殖、迁移和侵袭,但对胰腺癌干细胞生物学行为的影响及作用机制还未知。目的:探讨LINC02532对胰腺癌干细胞增殖、迁移、侵袭和凋亡的影响及作用机制。方法:①选取2017年1月至2018年6月于华北理工大学附属医院经病理检测证实为胰腺癌且经手术切除的56例患者胰腺癌组织及癌旁组织,实时荧光定量PCR(RT-qPCR)检测LINC02532和miR-145的表达水平;②以CD24+CD44+ESA+为表面标记,流式细胞术在人胰腺癌细胞PANC-1中分选胰腺癌干细胞,分为正常组、si-LINC02532组(转染LINC02532小干扰RNA)、si-NC组(转染乱序无意义阴性序列)、si-LINC02532+anti-miR-145组(共转染LINC02532小干扰RNA和miR-145抑制剂)和si-LINC02532+anti-miR-NC组(共转染LINC02532小干扰RNA和miR-145抑制剂阴性对照序列)。转染12 h后,RT-qPCR检测各组细胞中LINC02532和miR-145表达水平,CCK-8法检测细胞存活率,克隆形成实验检测细胞克隆形成数,Transwell小室检测细胞迁移和侵袭能力,流式细胞术检测细胞凋亡率,Western blot法检测细胞中P21、Bax、Caspases-3、E-钙黏附素和基质金属蛋白酶2蛋白表达水平。双荧光素酶报告基因实验验证miR-145与LINC02532调控关系。结果与结论:①与癌旁组织比较,胰腺癌组织中LINC02532表达水平升高(P<0.05),miR-145表达水平降低(P<0.05);②与si-NC组比较,si-LINC02532组胰腺癌干细胞存活率、克隆形成数、细胞迁移和侵袭数及基质金属蛋白酶2蛋白表达水平降低(P<0.05),细胞凋亡率及P21、Bax、Caspase-3和E-钙黏附素蛋白表达水平升高(P<0.05),而si-NC组与正常组各检测指标比较差异�BACKGROUND:Long non coding RNA(lncRNA)and microRNA(miRNA)are two types of small non-coding RNA that are involved in regulating cell proliferation,apoptosis and migration and other life activities,and lncRNA can also be used as a competitive endogenous RNA and miRNA target binding to regulate the expression of miRNA target genes and play an important role in the occurrence and development of tumors.LINC02532 is an lncRNA that can target miR-129-5p and miR-490-5p to promote the proliferation,migration and invasion of gastric cancer cells,but its effect on the biological behavior of pancreatic cancer stem cells and the action mechanism are still unknown.OBJECTIVE:To investigate the effect and action mechanism of LINC02532 on the proliferation,migration,invasion and apoptosis of pancreatic cancer stem cells.METHODS:(1)Fifty-six patients with pancreatic cancer confirmed by pathological examination in North China University of Science and Technology Affiliated Hospital from January 2017 to June 2018 were selected as the research subjects.Real time fluorescent quantitative PCR was used to detect the expression levels of LINC02532 and miR-145 in pancreatic cancer tissues and adjacent tissues.(2)Using CD24+CD44+ESA+as surface markers,pancreatic cancer stem cells were sorted from human pancreatic cancer cells PANC-1 by flow cytometry.Pancreatic cancer stem cells were divided into normal group,si-LINC02532 group(transfected with LINC02532 small interfering RNA),si-NC group(transfected with disorderly negative sequence),si-LINC02532+anti-miR-145 group(co-transfected with LINC02532 small interfering RNA and miR-145 inhibitor)and si-LINC02532+anti-miR-NC group(co-transfected with LINC02532 small interfering RNA and miR-145 inhibitor negative control sequence).At 12 hours after transfection,real time fluorescent quantitative PCR was used to detect the expression levels of LINC02532 and miR-145 in each group of cells.Cell counting kit-8(CCK-8)assay was utilized to detect the cell survival rate.The colony formation test was appli

关 键 词：LINC02532 MIR-145 胰腺癌干细胞 生物学行为 因子 蛋白 通路 靶向 凋亡 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]