LncRNA SNHG15/miR-123/PAK5轴通过自噬对胃癌细胞活性及血管生成的影响  

Mechanism of LncRNA SNHG15/miR⁃123/PAK5 axis regulating autophagy on gastric cancer cell activity and angiogenesis

作　　者：刘杰 刘树青 梁凤[2] 王蕾 刘永健 LIU Jie;LIU Shu-qing;LIANG Feng;WANG Lei;LIU Yong-jian(Huai'an Hospital Affiliated to Xuzhou Medical University,Huai'an,Jiangsu Province 223002,China;Department of Gastroenterology,Second People's Hospital of Huai'an City,Jiangsu Province 223002,China;Department of On-cology,Huai'an Fifth People's Hospital,Jiangsu Province 223300,China;Department of Gastrointestinal Surgery,Huai'an Fifth People's Hospital,Jiangsu Province 223002,China)

机构地区：[1]徐州医科大学附属淮安医院,江苏淮安223002 [2]淮安市第二人民医院消化科,江苏淮安223002 [3]淮安市第五人民医院肿瘤科,江苏淮安223300 [4]淮安市第五人民医院胃肠外科,江苏淮安223002

出　　处：《解剖学研究》2023年第3期242-250,共9页Anatomy Research

基　　金：淮安市卫生健康科研项目(HAWJ201916)。

摘　　要：目的探讨lncRNA SNHG15/miR-123/PAK5轴通过调节自噬对胃癌细胞活性及血管生成的机制研究。方法将胃癌SCG-823细胞分为Control组、si-NC组、si-SNHG15组、miR-NC组、miR-123组、pcDNAPAK5组。RT-PCR检测细胞中SNHG15和miR-123表达;MTT法检测细胞增殖能力;Transwell小室法检测细胞侵袭能力;流式细胞仪检测细胞凋亡能力;Matrigel体外成管实验检测细胞血管生成能力;蛋白质印迹检测细胞中PAK5、VEGF、LC3、Beclin1、p62蛋白表达;双荧光素酶报告基因检验SNHG15和miR-123、miR-123和PAK5的靶向关系。结果与人胃黏膜细胞系GES-1相比,人胃癌细胞系中ASG、MKN-45、SCG-823细胞中SNHG15表达明显升高,miR-123表达明显降低(P<0.05);且SCG-823细胞中SNHG15表达明显高于ASG、MKN-45细胞,miR-123表达明显低于ASG、MKN-45细胞(P<0.05)。si-SNHG15组细胞增殖、侵袭及形成小管数量均明显低于Control组,细胞凋亡率高于Control组(P<0.05);si-SNHG15组细胞中VEGF、PAK5、p62蛋白表达明显低于Control组,LC3Ⅱ/LC3Ι比值及Beclin1蛋白表达明显高于Control组(P<0.05)。与miR-NC组相比,miR-123组细胞增殖、侵袭及形成小管数量均明显降低,细胞凋亡率明显增加(P<0.05);miR-123组细胞中VEGF、PAK5、p62蛋白表达明显低于miR-NC组组,LC3Ⅱ/LC3Ι比值及Beclin1蛋白表达明显高于miRNC组(P<0.05)。通过向细胞中分别转染野生型SNHG15(SNHG15-WT)、PAK5(SNHG15-WT)时,miR-123组荧光素酶活性均明显低于miR-NC组(P<0.05)。与si-SNHG15组相比,pcDNA-PAK5组细胞细胞增殖、侵袭及形成小管数量均明显升高,细胞凋亡率明显降低(P<0.05);pcDNA-PAK5组细胞中VEGF、PAK5、p62蛋白表达明显高于si-SNHG15组,LC3Ⅱ/LC3Ι比值及Beclin1蛋白表达明显低于si-SNHG15组(P<0.05)。结论lncRNA SNHG15可靶向miR-123/PAK5轴抑制胃癌细胞增殖、侵袭和血管生成,促进胃癌细胞凋亡和自噬,进而为调控自噬途径治疗胃癌提供新的思路。Objective To investigate the mechanism of lncRNA SNHG15/miR⁃123/PAK5 axis regulating autophagy on gastric cancer cell activity and angiogenesis.Methods SCG⁃823 cells were divided into control group,si NC group,si SNHG15 group,miR⁃NC group,miR⁃123 group,pcDNA⁃PAK5 group.The expression of SNHG15 and miR⁃123 was detected by RT⁃PCR;MTT assay was used to detect cell proliferation;Transwell chamber method was used to detect the invasive ability of cells;Apoptosis was detected by flow cytometry;The angiogenesis ability of cells was detected by Matrigel tube formation experiment in vitro;Western blot was used to detect the expression of PAK5,VEGF,LC3,Beclin1 and p62 proteins in cells;Double luciferase reporter gene was used to test the targeting relationship between SNHG15 and miR⁃123,miR⁃123 and PAK5.Results Compared with human gastric mucosal cell line GES⁃1,the expression of SNHG15 in human gastric cancer cell line ASG,MKN⁃45,SCG⁃823 was signifi⁃cantly increased,and the expression of miR⁃123 was significantly decreased(P<0.05);The expression of SNHG15 in SCG⁃823 cells was significantly higher than that in ASG and MKN⁃45 cells,and the expression of miR⁃123 was significantly lower than that in ASG and MKN⁃45 cells(P<0.05).Compared with control group,the number of cell proliferation,invasion and tubule formation in si⁃SNHG15 group decreased significantly,and the apoptosis rate in⁃creased significantly(P<0.05);The expression of VEGF,PAK5,p62 protein in si⁃SNHG15 group was significantly lower than that in control group,LC3Ⅱ/LC3ΙThe ratio and the expression of Beclin1 protein in the control group were significantly higher than those in the control group(P<0.05).Compared with miR⁃NC group,the number of cell proliferation,invasion and tubule formation in miR⁃123 group decreased significantly,and the apoptosis rate increased significantly(P<0.05);The expression of VEGF,PAK5,p62 protein in miR⁃123 group was significantly lower than that in miR⁃NC group,LC3Ⅱ/LC3ΙThe ratio a

关 键 词：人胃癌细胞株 血管生成 自噬 lncRNA SNHG15/miR-123/PAK5轴 小核仁RNA宿主基因15 增殖 

分 类 号：R735.2[医药卫生—肿瘤]