lncRNA NNT-AS1调控miR-518a-3p抑制IL-17诱导胃癌细胞增殖、迁移和侵袭的分子机制  

Long noncoding RNA NNT-AS1 inhibits IL-17-induced proliferation,migration,and invasion of gastric cancer cells by regulating miR-518a-3p

作　　者：张晶 王博[3] 王红霞[2] 孔凡铭[1] 李小江[1] 贾英杰[1] Jing Zhang;Bo Wang;Hong-Xia Wang;Fan-Ming Kong;Xiao-Jiang Li;Ying-Jie Jia(Department of Oncology,The First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300380;Department of Gastroenterology,The First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300380;Department of Emergency Medicine,Tianjin Third Central Hospital,Tianjin 300380)

机构地区：[1]天津中医药大学第一附属医院肿瘤科,天津市300380 [2]天津中医药大学第一附属医院消化科,天津市300380 [3]天津市第三中心医院急诊科,天津市300380

出　　处：《世界华人消化杂志》2022年第13期571-578,共8页World Chinese Journal of Digestology

基　　金：国家自然科学基金,No.81503392.

摘　　要：背景白细胞介素17(interleukin 17,IL-17)是多细胞分泌因子,与肿瘤细胞发展有关,lncRNA NNT-AS1胃癌组织中lncRNA NNT-AS1表达上调,lncRNA NNT-AS1可能通过降低miR-363的表达抑制胃癌细胞增殖和侵袭,并阻滞细胞周期的发展.关于lncRNA NNT-AS1与IL-17调控作用对胃癌细胞研究尚不明确.目的探究长链非编码RNA烟酰胺核苷酸转氢酶反义RNA1(lncRNA NNT-AS1)调控miR-518a-3p抑制IL-17诱导胃癌细胞增殖、迁移和侵袭的分子机制.方法使用IL-17处理胃癌细胞AGS,将细胞分为对照(Con)组、IL-17组、IL-17+si-NC组、IL-17+si-lncRNA NNT-AS1组、IL-17+si-lncRNA NNT-AS1+anti-miR-NC组、IL-17+si-lncRNA NNT-AS1+anti-miR-518a-3p组.实时荧光定量PCR(RT-qPCR)检测miR-518a-3p和lncRNA NNT-AS1表达水平;克隆实验、Transwell检测AGS细胞增殖、迁移、侵袭;蛋白印迹法(Western blot)检测增殖、转移蛋白表达;双荧光素酶实验检测miR-518a-3p和lncRNA NNT-AS1关系.结果与Con组相比,IL-17组克隆细胞数、迁移细胞数、侵袭细胞数增多,lncRNA NNT-AS1表达水平、细胞核增殖抗原标记物(Ki-67)、基质金属蛋白酶2(matrix metalloproteinase 2,MMP2)、神经钙黏蛋白(N-cadherin)蛋白表达升高,miR-518a-3p表达水平、上皮钙黏蛋白(E-cadherin)蛋白表达降低.下调lncRNA NNT-AS1降低IL-17处理胃癌细胞克隆细胞数、迁移细胞数、侵袭细胞数,下调Ki-67、N-cadherin、MMP2蛋白表达,上调E-cadherin蛋白表达增加.lncRNA NNT-AS1靶向调控miR-518a-3p的表达,抑制miR-518a-3p可以逆转下调NNT-AS1对IL-17诱导胃癌细胞增殖、迁移和侵袭的影响.结论lncRNA NNT-AS1靶向调控miR-518a-3p抑制IL-17诱导胃癌细胞增殖、迁移和侵袭.BACKGROUND Interleukin 17(IL-17)is a cytokine secreted in many cell types,which is related to the development of tumor cells.The expression of the long noncoding RNA(lncRNA)nicotinamide nucleotide transhydrogenase antisense RNA1(NNT-AS1)is up-regulated in gastric cancer tissues.NNT-AS1 may inhibit the proliferation and invasion of gastric cancer cells and arrest cell cycle progression by reducing the expression of miR-363.The regulatory mechanism of NNT-AS1 and IL-17 on gastric cancer cells is still not completely clear.AIM To explore the molecular mechanism by which NNT-AS1 inhibits IL-17-induced proliferation,migration,and invasion of gastric cancer cells.METHODS IL-17 was used to treat gastric cancer AGS cells,and the cells were divided into Con group,IL-17 group,IL-17+si-NC group,IL-17+si-lncRNA NNT-AS1 group,IL-17+si-lncRNA NNT-AS1+anti-miR-NC group,and IL-17+si-lncRNA NNT-AS1+anti-miR-518a-3p group.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect miR-518a-3p and NNT-AS1 expression.Colony forming assay and Transwell assay were performed to detect AGS cell proliferation,migration,and invasion.Western blot was used to detect relevant protein expression.Dual luciferase assay was used to detect the relationship of miR-518a-3p and NNT-AS1.RESULTS Compared with the Con group,the number of cell colonies,the number of migrating cells,and the number of invasive cells in the IL-17 group increased,the expression of Ki-67,N-cadherin,and MMP2 proteins increased,the expression of E-cadherin protein decreased,the expression of NNT-AS1 increased,and the expression of miR-518a-3p decreased.Down-regulation of NNT-AS1 decreased the number of cell colonies formed,the number of migrating cells,and the number of invasive cells in gastric cancer cells treated with IL-17,decreased the expression of Ki-67,matrix metalloproteinase 2(MMP2),and N-cadherin proteins,and increased the expression of E-cadherin protein.NNT-AS1 targets and regulates the expression of miR-518a-3p.Inhibition of miR-518a-3p can reverse the eff

关 键 词：胃癌 增殖 迁移 侵袭 

分 类 号：R735.2[医药卫生—肿瘤]