LncRNA ASB16-AS1调节miR-185-5p/TRAM2轴对口腔鳞癌细胞增殖、迁移和侵袭的影响  

Effects of LncRNA ASB16-AS1 on the Proliferation,Migration,and Invasion of Oral Squamous Cell Carcinoma Cells by Regulating the miR-185-5p/TRAM2 Axis

作　　者：靳瑞 焦合主 黄颖[2] 张敏[2] JIN Rui;JIAO Hezhu;HUANG Ying;ZHANG Min(Department of Stomatology,Chinese People’s Liberation Army Airborne Troops Hospital,Wuhan 430000,China;Department of Outpatient,Chinese People’s Liberation Army Airborne Troops Hospital,Wuhan 430000,China)

机构地区：[1]中国人民解放军空降兵部队医院口腔科,武汉430000 [2]中国人民解放军空降兵部队医院机关门诊部,武汉430000

出　　处：《中国细胞生物学学报》2024年第7期1432-1441,共10页Chinese Journal of Cell Biology

摘　　要：该文旨在探讨长链非编码RNA ASB16-AS1(LncRNA ASB16-AS1)靶向微小RNA-185-5p(miR-185-5p)/易位相关膜蛋白2(TRAM2)轴对口腔鳞状细胞癌(又称口腔鳞癌)细胞增殖、迁移和侵袭的影响。体外培养人口腔鳞癌细胞SCC-9,将其分为对照组、sh-NC组、sh-ASB16-AS1组、sh-ASB16-AS1+inhibitor NC组、sh-ASB16-AS1+miR-185-5p inhibitor组。用qRT-PCR法检测各组细胞LncRNA ASB16-AS1、miR-185-5p、TRAM2 mRNA表达水平;用CCK-8试剂盒检测SCC-9细胞增殖情况;用流式细胞术检测SCC-9细胞凋亡情况;用划痕实验检测SCC-9细胞迁移情况;用Transwell法检测SCC-9细胞侵袭情况;Western blot检测TRAM2蛋白表达水平。双荧光素酶报告实验验证LncRNA ASB16-AS1与miR-185-5p及miR-185-5p与TRAM2之间的靶向关系。与对照组和sh-NC组相比,sh-ASB16-AS1组中Lnc RNA ASB16-AS1、TRAM2 mRNA、TRAM2蛋白表达水平以及D值、划痕愈合率、细胞侵袭数降低,mi R-185-5p水平、细胞凋亡率升高(P<0.05);与sh-ASB16-AS1+inhibitor NC组相比,sh-ASB16-AS1+miR-185-5p inhibitor组中mi R-185-5p水平、细胞凋亡率降低,TRAM2 m RNA表达水平、TRAM2蛋白表达水平、D值、划痕愈合率、细胞侵袭数升高(P<0.05),而Lnc RNA ASB16-AS1水平无显著变化(P>0.05);生物学信息网站预测显示mi R-185-5p与Lnc RNA ASB16-AS1和TRAM2均存在靶向结合位点,且双荧光素酶报告基因实验证实LncRNA ASB16-AS1与miR-185-5p,以及miR-185-5p与TRAM2之间均存在靶向关系(P<0.05)。在口腔鳞癌细胞中Lnc RNA ASB16-AS1表达上调,抑制Lnc RNA ASB16-AS1的表达可靶向上调mi R-185-5p的表达,抑制TRAM2的表达,进而促进口腔鳞癌细胞凋亡,抑制口腔鳞癌细胞增殖、迁移和侵袭。This study aims to investigate the effects of LncRNA ASB16-AS1(long non-coding RNA ASB16-AS1)on the proliferation,migration,and invasion of oral squamous cell carcinoma cells by targeting the miR-185-5p/TRAM2 axis.Human oral squamous cell carcinoma cells SCC-9 were cultured in vitro and grouped into control group,sh-NC group,sh-ASB16-AS1 group,sh-ASB16-AS1+inhibitor NC group,and sh-ASB16-AS1+miR-185-5p inhibitor group.qRT-PCR method was applied to detect the expression of LncRNA ASB16-AS1,miR-185-5p,and TRAM2 mRNA of cells in each group.SCC-9 cell proliferation was detected using CCK-8 assay kit.SCC-9 cell apoptosis was detected by flow cytometry.SCC-9 cell migration was detected using scratch assay,and SCC-9 cell invasion was detected using Transwell method.Western blot was applied to detect the expression of TRAM2 protein.Dual luciferase reporter assay was applied to verify the targeting relationship between LncRNA ASB16-AS1 and miR-185-5p,and miR-185-5p and TRAM2.Compared with the control group and sh-NC group,the expression levels of LncRNA ASB16-AS1,TRAM2 mRNA,and TRAM2 protein,as well as D value,scratch healing rate,and number of invasive cells were reduced in the sh-ASB16-AS1 group,while the level of miR-185-5p and cell apoptosis rate were increased(P<0.05).Compared with the sh-ASB16-AS1+inhibitor NC group,the miR-185-5p level and cell apoptosis rate in the sh-ASB16-AS1+miR-185-5p inhibitor group were reduced,while the ex-pression levels of TRAM2 mRNA and TRAM2 protein,D values,scratch healing rate,and number of invasive cells were elevated(P<0.05),however,there were no significant changes in the levels of LncRNA ASB16-AS1(P>0.05).Biological information websites predicted that miR-185-5p had targeted binding sites with LncRNA ASB16-AS1 and TRAM2,and dual luciferase reporter gene experiments confirmed the existence of targeted relationships be-tween LncRNA ASB16-AS1 and miR-185-5p,and miR-185-5p and TRAM2(P<0.05).The expression of LncRNA ASB16-AS1 is upregulated in oral squamous cell carcinoma cells.Inhibit

关 键 词：长链非编码RNA ASB16-AS1 微小RNA-185-5p/易位相关膜蛋白2 口腔鳞癌 增殖 迁移 侵袭 

分 类 号：R739.8[医药卫生—肿瘤]