长链非编码RNA 1010001N08Rik和GATA6在高氧暴露小鼠肺癌细胞中表达和意义  

Expression and Significance of Long non-coding RNA 1010001N08Rik and GATA 6 in Hyperoxia-exposed LLC Cells

作　　者：罗好 刘怡波 颜崇兵 翁博雯 蔡成 LUO Hao;LIU Yi-bo;YAN Chong-bing;WENG Bo-wen;CAI Cheng(Department of Neonatology,School of medicine,Shanghai JiaoTong University,Shanghai Children's Hospital,Shanghai,200062,China)

机构地区：[1]上海交通大学医学院附属儿童医院上海市儿童医院新生儿科,上海200062

出　　处：《现代生物医学进展》2024年第15期2805-2816,共12页Progress in Modern Biomedicine

基　　金：2023年"科技兴蒙"上海交通大学行动计划专项项目;2023年上海市儿童医院院级临床研究培育专项(2023YLY02)。

摘　　要：目的:探讨长链非编码RNA(long noncoding RNA,lncRNA)1010001N08Rik和GATA6在高氧暴露小鼠肺癌细胞(Lewis lung carcinoma cell,LLC细胞)中表达量的变化与相关性,及其介导Wnt/β-catenin信号通路参与高氧肺损伤的机制。方法:(1)将LLC细胞分为空气组、高氧暴露24 h组和高氧暴露48h组,应用RT-PCR和Western blot法分别检测lncRNA1010001N08Rik、GATA6、Wnt及β-catenin的m RNA及相关蛋白表达水平。(2)将LLC细胞分为无干扰空气组、无干扰高氧组、NC-si RNA干扰高氧组、si RNA干扰高氧组。应用RT-PCR和Western blot法分别检测lncRNA1010001N08Rik、GATA6、Wnt及β-catenin的m RNA及相关蛋白表达水平,流式细胞术检测各组细胞凋亡。(3)利用GraphPad Prism 9.4.1进行数据可视化。(4)采用SPSS 26.0统计软件进行数据处理,两组间数据比较采用两独立样本t检验,多组间比较采用单因素方差分析。(5)将Wnt、Fzd2、GATA6等相关基因输入string11.0数据库构建PPI网络并导出PPI网络图。结果:(1)与空气组相比较,高氧暴露24 h组和48h组GATA6的m RNA表达量下降,lncRNA1010001N08Rik、Wnt、β-catenin的m RNA表达量增加,差异具有统计学意义(P<0.05)。(2)与空气组相比,高氧暴露24 h组和48 h组GATA6蛋白表达量下降,Wnt、β-catenin蛋白表达量增加,差异具有统计学意义(P<0.05)。(3)与无干扰空气组相比,无干扰高氧组、NC-si RNA干扰高氧组的GATA6的m RNA表达量和蛋白表达量下降,1010001N08Rik的m RNA表达量、Wnt、β-catenin的m RNA表达量和蛋白表达量增加;无干扰高氧组与NC-si RNA干扰高氧组的比较差异无统计学意义;与干扰高氧组与NC-si RNA干扰高氧组相比,si RNA干扰高氧组GATA6的m RNA表达量和蛋白表达量增加,1010001N08Rik的m RNA表达量、Wnt、β-catenin的m RNA表达量和蛋白表达量下降,差异均具有统计学意义(P<0.05)。(4)与无干扰空气组相比,无干扰高氧组、NC-si RNA干扰高氧组细胞凋亡率增加;无干扰高氧组Objective:To investigate the expression changes and correlation of long non-coding RNA(lncRNA)1010001N08Rik and GATA6 in hyperoxic LLC cells,and the mechanism of Wnt/β-catenin signaling pathway involved in hyperoxic lung injury.Method:(1)LLC cells were divided into air group.The expression levels of GATA6,Wnt,β-catenin,1010001N08Rik in each group were detected by RT-PCR and the expression levels of related proteins in each group were detected by Western blot.(2)LLC cells were divided into non-interference air group,non-interference hyperoxia group,NC-siRNA interference hyperoxia group and siRNA interference hyperoxia group.The expression levels of GATA6,Wnt,β-catenin,1010001N08Rik in each group were detected by RT-PCR and the expression levels of related proteins in each group were detected by Western blot.The apoptosis was detected by flow cytometry.(3)Data visualization using GraphPad Prism 9.4.1.(4)SPSS 26.0 statistical software was used for data processing.The data of two groups were compared by t test.One-way analysis of variance was used for comparison among multiple groups.(5)Wnt,Fzd2,GATA6 and other related genes were input into string11.0 database to construct the PPI network and derive the PPI network diagram.Results:(1) Compared with the air group, the expression of GATA6 mRNA decreased in the hyperoxia exposure group for 24 h and hyperoxiaexposure group for 48h, the expression of lncRNA1010001N08Rik mRNA, Wnt mRNA, β-catenin mRNA expression increased. Thedifference was statistically significant (P<0.05). (2) Compared with the air group, the expression of GATA6 protein decreased in thehyperoxia exposure group for 24 h and hyperoxia exposure group for 48 h, whlie the expression of Wnt protein, β-catenin proteinincreased. The difference was statistically significant (P<0.05). (3) Compared with the non-interference air group, the expression ofGATA6 mRNA and protein in the non-interference hyperoxia group and the NC-siRNA interference hyperoxia group decreased, and theexpression of 1010001N08Rik mRNA,

关 键 词：高氧暴露 lncRNA1010001N08Rik GATA6 高氧肺损伤 WNT/Β-CATENIN信号通路 

分 类 号：R-33[医药卫生] R722[生物学—生物工程] Q813