LncRNA PXN-AS1对胃癌细胞增殖、凋亡及侵袭的影响  

Effects of LncRNA PXN-AS1 on proliferation,apoptosis and invasion of gastric cancer cells

作　　者：刘刚[1] 刘东涛[1] 赵伟[1] 何涛[1] 张媛 Liu Gang;Liu Dongtao;Zhao Wei;He Tao;Zhang Yuan(Dept of Gastroenterology,General Hospital of Ningxia Medical University,Yinchuan 750004)

机构地区：[1]宁夏医科大学总医院胃肠外科,银川750004

出　　处：《安徽医科大学学报》2025年第3期430-439,共10页Acta Universitatis Medicinalis Anhui

基　　金：宁夏回族自治区科技厅重点研发计划项目(编号:2022BEG03154)。

摘　　要：目的 探讨LncRNA PXN-AS1(简称PXN-AS1)在胃癌组织中的表达水平及其对胃癌细胞增殖、凋亡及侵袭的影响,并阐述其可能的调控机制。方法 收集43例胃癌患者的胃癌组织及癌旁组织,采用qRT-PCR法检测胃癌组织中PXN-AS1和miR-125a-5p表达水平,并分析PXN-AS1表达水平与胃癌患者临床病理参数的关系;采用Pearson法分析胃癌组织PXN-AS1和miR-125a-5p的相关性。将PXN-AS1 siRNA质粒(si-PXN-AS1)及阴性对照质粒(si-NC)与miR-125a-5p inhibitor(inhibitor)及阴性对照miRNA抑制剂(inhibitor NC)分别或同时共转染至HGC-27细胞中,分为si-NC组、si-PXN-AS1组、si-PXN-AS1+inhibitor NC组、si-PXN-AS1+inhibitor组,另设立空白对照组(blank组)。采用qRT-PCR法检测各组细胞中PXN-AS1和miR-125a-5p表达水平;CCK-8法检测各组细胞的增殖水平;EdU实验检测各组细胞增殖情况;Transwell实验检测各组细胞迁移及侵袭能力;流式细胞术检测各组细胞的凋亡水平;双荧光素酶报告基因系统验证PXN-AS1海绵吸附miR-125a-5p并调控其表达水平。应用生物信息学筛选miR-125a-5p下游靶基因,并进行GO和KEGG富集分析。结果 与癌旁组织比较,胃癌组织中PXN-AS1表达水平明显升高,而miR-125a-5p表达水平明显降低(均P<0.05),PXN-AS1和miR-125a-5p呈负相关(r=-0.452,P=0.002)。此外,PXN-AS1高表达的胃癌患者淋巴结转移阳性和低分化比例明显增高(均P<0.05)。与blank组和si-NC组比较,si-PXN-AS1组HGC-27细胞中PXN-AS1表达水平、细胞增殖能力、细胞迁移和侵袭数量明显降低(均P<0.05),而细胞凋亡水平明显升高(P<0.05)。双荧光素酶报告系统结果显示,在HGC-27细胞中PXN-AS1特异性吸附miR-125a-5p。敲低miR-125a-5p可逆转沉默PXN-AS1对胃癌细胞增殖、迁移和侵袭的抑制作用,对细胞凋亡的促进作用。生信分析结果显示,miR-125a-5p的下游靶基因可以参与多种生物过程,包括DNA损伤、T细胞凋亡和分化、RNA代谢合成过程及信号转导Objective To investigate the expression of LncRNA PXN-AS1(abbreviated as PXN-AS1)in gastric cancer tissues and its impact on the proliferation,apoptosis,and invasion of gastric cancer cells,and to explore its potential regulatory mechanisms.Methods Gastric cancer tissues and adjacent tissues from 43 patients with gastric cancer were collected.The expression levels of PXN-AS1 and miR-125a-5p in gastric cancer tissues were detected using qRT-PCR,and the relationship between the expression level of PXN-AS1 and the clinical pathological parameters of patients with gastric cancer was analyzed.Pearson correlation analysis was conducted to evaluate the correlation between PXN-AS1 and miR-125a-5p in gastric cancer tissues.PXN-AS1 siRNA plasmid(si-PXN-AS1)and negative control plasmid(si-NC),as well as miR-125a-5p inhibitor(inhibitor)and negative control miRNA inhibitor(inhibitor NC),were transfected alone or in combination into HGC-27 cells.The cells were divided into si-NC group,si-PXN-AS1 group,si-PXN-AS1+inhibitor NC group,si-PXN-AS1+inhibitor group,and a blank control group was also established.qRT-PCR was utilized to assess the expression levels of PXN-AS1 and miR-125a-5p in each cell group.CCK-8 assay was employed to evaluate the proliferation levels of cells in each group.EdU experiment was conducted to assess the proliferation status of cells in each group.Transwell assay was performed to examine the migration and invasion capabilities of cells in each group;flow cytometry was employed to measure the apoptosis levels of cells in each group;the dual-luciferase reporter gene system was used to validate the sponge absorption of PXN-AS1 on miR-125a-5p and regulate its expression levels.Bioinformatics analysis was employed to screen downstream target genes of miR-125a-5p and perform GO and KEGG enrichment analysis.Results Compared with adjacent cancerous tissues,the expression level of the PXN-AS1 in gastric cancer tissues significantly increased,while the expression level of miR-125a-5p markedly decreased(both P<0.05)

关 键 词：LncRNA PXN-AS1 胃癌 增殖 迁移和侵袭 PXN-AS1/miR-125a-5p轴 

分 类 号：R735.2[医药卫生—肿瘤]