过表达lncRNA ATG16L2-211通过促进lncRNA GAS6-AS1表达抑制肺腺癌细胞的生长和迁移  

LncRNA ATG16L2-211 overexpression inhibits growth and migration of lung adenocarcinoma cells by promoting lncRNA GAS6-AS1 expression

作　　者：王敏[1] 李刚[1] 程小彬 李晶 汪六林 龙琦 Wang Min;Li Gang;Cheng Xiaobin;Li Jing;Wang Liulin;Long Qi(Department of Critical Care Medicine,Hubei Provincial Hospital of Traditional Chinese Medicine,Wuhan 430061,China)

机构地区：[1]湖北省中医院重症医学科,武汉430061

出　　处：《国际呼吸杂志》2022年第1期50-55,共6页International Journal of Respiration

基　　金：湖北省卫生健康委联合基金项目(WJ2019H172)。

摘　　要：目的分析长链非编码RNA(lncRNA)ATG16L2-211在肺腺癌组织和细胞株中的表达, 研究其对肺腺癌细胞生长和迁移的影响及其分子机制。方法本研究为实验室研究。GEPIA在线数据库分析ATG16L2-211在肺腺癌组织中的表达情况。采用实时定量聚合酶链式反应检测ATG16L2-211在肺腺癌细胞株(H1650、A549、H1975、H1299)中的表达情况。将ATG16L2-211序列和阴性对照序列转入H1650细胞, 分别标记为ATG16L2-211组和阴性对照组。CCK-8法检测H1650细胞活力, 细胞划痕实验检测H1650细胞迁移。GEPIA在线数据库分析ATG16L2-211相关性较高的基因。实时定量聚合酶链式反应和Western blot检测ATG16L2-211相关基因的表达。结果 ATG16L2-211在肺腺癌组织中表达低于正常组织(t=48.12, P<0.001)。ATG16L2-211在肺腺癌细胞株中表达均低于正常肺泡上皮细胞(P值均<0.05), H1650细胞中ATG16L2-211表达最低(F=13.79, P<0.001)。与阴性对照组比较, 从2 d开始至5 d ATG16L2-211组H1650细胞增殖活力均降低(P值均<0.05)。阴性对照组和ATG16L2-211组划痕愈合率为(72.15±6.23)%和(21.54±4.08)%, ATG16L2-211组H1650细胞迁移能力降低(t=6.79, P=0.001)。肺腺癌组织中ATG16L2-211和GAS6-AS1表达呈显著正相关(r=0.60, P<0.001)。与阴性对照组比较, ATG16L2-211组H1650细胞中GAS6-AS1表达增加(t=3.37, P=0.015), 葡萄糖转运蛋白1基因表达降低(t=4.33, P=0.005), 转化生长因子β1信号通路蛋白表达降低。结论 ATG16L2-211在肺腺癌组织及细胞株中低表达, 上调ATG16L2-211能通过促进GAS6-AS1表达发挥抑制肺腺癌细胞生长和迁移的作用。Objective To analyze the expression of autophagy related 16 like 2(ATG16L2)-211 of long non-coding RNA(lncRNA)in lung adenocarcinoma(LUAD)tissue and cell lines to study effect on growth and migration of LUAD cells and its molecular mechanism.Methods This is a laboratory study.The gene expression profiling interactive analysis(GEPIA)database was used to analyze the expression of ATG16L2-211 in LUAD tissues.The expression of ATG16L2-211 in LUAD cell lines(H1650,A549,H1975,H1299)was detected by quantitative real-time polymerase chain reaction(PCR).ATG16L2-211 sequence and the negative control(NC)sequence were transferred into H1650 cells and labeled as ATG16L2-211 group and NC group.Human lung cancer cells(H1650 cells)viability and H1650 cells migration were detected with cholecystokinin-octapeptide(CCK-8)method and cell wound scratch assay,respectively.Meanwhile,analyzing genes in closely related ATG16L2-211with GEPIA database,predicting ATG16L2-211-related gene with quantitative real-time PCR and Western blot.Results The expression of ATG16L2-211 in LUAD tissues was lower than that in normal tissues(t=48.12,P<0.001),which was lower in LUAD cell lines than in Human Pulmonary Alveolar Epithelial Cells(HPAEpiC)(all P<0.05),and it was the lowest in H1650 cells(F=13.79,P<0.001).H1650 cells viability from day 2 to day 5 in ATG16L2-211 group were decreased in comparison with NC group(all P<0.05).The wound healing rate was(72.15±6.23)% and(21.54±4.08)%in NC group and ATG16L2-211 group,respectively,and H1650 cells migration in ATG16L2-211 group was decreased(t=6.79,P=0.001).The expression of ATG16L2-211 in LUAD tissue was positively correlated with GAS6 antisense RNA 1(GAS6-AS1)(r=0.60,P<0.001).Increased expression of GAS6-AS1 in H1650 cells(t=3.37,P=0.015),decreased expression of glucose transporter-1(GLUT-1)(t=4.33,P=0.005),decreased protein expression of transforming growth factorβ1(TGF-β1)signaling pathway were detected in ATG16L2-211 group as compared to NC group.Conclusions The expression of ATG16L2-211 is low in

关 键 词：肺肿瘤 腺癌 细胞增殖 细胞转移 ATG16L2-211 GAS6-AS1 

分 类 号：R734.2[医药卫生—肿瘤]