摘要
目的探究长链非编码RNA转移相关肺腺癌转录本1(lnc-MALAT1)在脂多糖(LPS)诱导的大鼠肺泡巨噬细胞系(AMOs)炎性反应中的调控机制。方法LPS处理AMOs建立细胞损伤模型,用脂质体法分别将pcDNA、pcDNA-MALAT1、miR-NC、miR-217 mimics、pcDNA-MALAT1与miR-NC、pcDNA-MALAT1与miR-217 mimics转染至AMOs,LPS处理12 h。采用酶联免疫吸附法(ELISA)检测细胞分泌人白介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)的量;采用RT-qPCR检测细胞中MALAT1、miR-217的表达;采用双荧光素酶报告实验、RNA结合蛋白免疫沉淀实验(RIP)检测MALAT1与miR-217的靶向关系。结果与AMOs组相比,LPS+AMOs组细胞中IL-1β、TNFα的含量显著升高(P<0.05),MALAT1表达水平显著升高,miR-217表达水平显著降低(P<0.05)。双荧光素酶报告实验与RIP实验证实MALAT1能够靶向结合miR-217。过表达MALAT1可明显促进LPS诱导的AMOs细胞IL-1β、TNF-α的分泌,而过表达miR-217则可抑制LPS诱导的AMOs细胞IL-1β、TNF-α的分泌,并且过表达miR-217减轻MALAT1促进LPS诱导的AMOs细胞IL-1β、TNF-α的分泌。结论Lnc-MALAT1可促进LPS诱导的AMOs的炎性反应,其机制与靶向miR-217相关。
Objective To investigate the regulatory mechanism of long-chain noncoding RNA metastasis-associated lung adenocarcinoma 1(lnc-MALAT1)in the inflammatory response of lipopolysaccharide(LPS)-induced alveolar macrophages(AMOs).Methods AMOs cells were treated with LPS for 12 hours to create an inflammation model,and liposome method was used to transfect pcDNA,pcDNA-MALAT1,miR-NC,miR-217 mimics,pcDNA-MALAT1 and miR-NC,pcDNA-MALAT1 and miR-217 mimics to AMOs.The level of IL-1βand TNF-αwas detected by ELISA.RT-qPCR was used to detect the expression of MALAT1 and miR-217 in cells.The dual luciferase report experiment and RIP experiment were used to detect potential target-attacking relationship between MALAT1 and miR-217.Results Compared with the AMOs group,the expression of IL-1βand TNFα,MALAT1 in the cells of the LPS+AMOs group were significantly increased(P<0.05)and theexpression level of miR-217 was significantly reduced(P<0.05).The dual luciferase report experiment and RIP experiment confirmed that MALAT1 targeted at miR-217.Over-expression of MALAT1 could obviously promote the secretion of IL-1βand TNF-αinduced by LPS in AMOs,while over-expression of miR-217 inhibited the secretion of IL-1βand TNF-αinduced by LPS in AMOs.Furthermore,over-expression of miR-217 reduced the secretion of IL-1βand TNF-αinduced by MALAT1 in LPS-induced AMOs.Conclusions Lnc-MALAT1,a long non-coding RNA,can promote the inflammatory response of AMOs induced by LPS,and its mechanism is related to its targeting at miR-217.
作者
罗传皊
孙利平
戚成栋
LUO Chuan-ling;SUN Li-ping;QI Cheng-dong(Department of Critical Medicine,Zaozhuang Mining Group Central Hospital,Zaozhuang 277100,China;Department of Pediatric Medicine, Zaozhuang Mining Group Central Hospital, Zaozhuang 277100,China)
出处
《基础医学与临床》
2021年第3期404-408,共5页
Basic and Clinical Medicine
