LncRNA FEZF1-AS1通过调控EZH2对肺间质细胞增殖、迁移及侵袭的作用

doi:10.16352/j.issn.1001-6325.2024.01.0043

基础医学与临床 ›› 2024, Vol. 44 ›› Issue (1): 43-50.doi: 10.16352/j.issn.1001-6325.2024.01.0043

LncRNA FEZF1-AS1通过调控EZH2对肺间质细胞增殖、迁移及侵袭的作用

王春燕¹, 王萍², 宋龙飞³, 刘永全^2*, 满君^2*

1.潍坊医学院临床医学院,山东潍坊 261035;
2.潍坊医学院附属医院呼吸内科,山东潍坊 261035;
3.潍坊医学院附属医院康复医学科,山东潍坊 261035

收稿日期:2023-07-14 修回日期:2023-11-07 出版日期:2024-01-05 发布日期:2023-12-25
通讯作者: ^*:manjun0229@126.com; xuanyuanlyq@163.com
基金资助:
国家自然科学基金青年科学基金(82205079)

Effects of lncRNA FEZF1-AS1 on proliferation, migration and invasion through regulating EZH2 of lung interstitial cells

WANG Chunyan¹, WANG Ping², SONG Longfei³, LIU Yongquan^2*, MAN Jun^2*

1. Clinical Medical College, Weifang Medical University, Weifang 261035;
2. Department of Respiratory, the Affiliated Hospital of Weifang Medical University,Weifang 261035, China;
3. Department of Rehabilitation Medicine, the Affiliated Hospital of Weifang Medical University,Weifang 261035, China

Received:2023-07-14 Revised:2023-11-07 Online:2024-01-05 Published:2023-12-25
Contact: ^*:manjun0229@126.com; xuanyuanlyq@163.com

摘要/Abstract

摘要： 目的研究长链非编码RNA FEZ家族锌指1-反义RNA 1(lncRNA FEZF1-AS1)调控zeste同源物增强子2(EZH2)对肺间质细胞增殖、迁移、侵袭能力及上皮细胞-间质转化(EMT)的影响及其作用机制。方法将人肺腺癌细胞系A549分为对照组(control)和模型组[model,用转化生长因子β1(TGF-β1) 20 ng/mL作用48 h,诱导成为肺间质细胞]。用Western blot检测细胞中E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)及波形蛋白(vimentin)的蛋白表达。RT-qPCR检测细胞中lncRNA FEZF1-AS1和EZH2基因表达。转染组细胞分为转染si NC组、si lncRNA FEZF1-AS1+OE vector组和si lncRNA FEZF1-AS1+OE EZH2组。CCK-8法检测细胞增殖、细胞划痕检测细胞迁移、Transwell小室法检测细胞侵袭;用Western blot检测细胞中E-cadherin、N-cadherin、vimentin及EZH2的蛋白表达,用RNA免疫沉淀(RIP)测定FEZF1-AS1与EZH2的直接结合作用。结果与对照组比较,模型组E-cadherin的蛋白表达水平减少(P＜0.05);N-cadherin及vimentin的蛋白表达水平升高(P＜0.05);与对照组比较,模型组lncRNA FEZF1-AS1与EZH2基因的表达水平明显升高(P＜0.05);与si NC组相比,si lncRNA FEZF1-AS1+OE vector组细胞增殖、迁移、侵袭能力降低,E-cadherin蛋白表达升高,N-cadherin、vimentin、EZH2蛋白表达降低(P＜0.05);与si lncRNA FEZF1-AS1+OE vector组比较,si lncRNA FEZF1-AS1+OE EZHZ组细胞增殖、侵袭、迁移能力升高,E-cadherin蛋白表达降低,N-cadherin、vimentin、EZH2蛋白表达升高(P＜0.05);RIP实验进一步证实了lncRNA FEZF1-AS1与EZH2具有结合作用。结论 LncRNA FEZF1-AS1通过调控EZH2促进肺间质细胞增殖、侵袭、转移和EMT过程。

关键词: 特发性肺间质纤维化, FEZ家族锌指1-反义RNA 1(FEZF1-AS1), 上皮细胞-间充质转化(EMT), zeste基因增强子同源物2(EZH2), 人非小细胞肺癌细胞系A549

Abstract: Objective To investigate the effects of long non-coding RNA FEZ family zinc finger 1 antisense RNA 1(lncRNA FEZF1-AS1) on enhancer of zeste homolog 2(EZH2) in regulation of proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of pulmonary interstitial cells and its mechanism. Methods The A549 cells human lung adenocarcinoma cell line were divided into control group and model group [model cells were induced into lung interstitial cells after being treated with transforming growth factor β1(TGF-β1)20 ng/mL for 48 h]. The protein expression of E-cadherin, N-cadherin and vimentin in each group was detected by Western blot. The expression of lncRNA FEZF1-AS1 and EZH2 in the two groups was detected by RT-qPCR. Cells in the transfection group were divided into si NC group,lncRNA FEZF1-AS1+OE vector group and si lncRNA FEZF1-AS1+OE EZH2 group. Cell proliferation was examined by CCK-8 method, cell migration was detected by cell scratch, and cell invasion was detected by Transwell assays. The protein expression of E-cadherin, N-cadherin, vimentin and EZH2 in each group was detected by Western blot. The direct binding effect of FEZF1-AS1 and EZH2 was determined by RNA immuno-precipitation (RIP). Results Compared with the control group, the protein expression level of E-cadherin in the model group was significantly decreased (P＜0.05),and the protein expression of N-cadherin and vimentin was significantly increased(P＜0.05). Compared with the control group, the expression level of lncRNA FEZF1-AS1 and EZH2 genes was significantly increased in the model group (P＜0.05). Compared with si NC group, the proliferation, migration and invasion ability of si lncRNA FEZF1-AS1+OE vector group were decreased, the expression of E-cadherin protein was increased while the expression of N-cadherin, vimentin and EZH2 was decreased(P＜0.05). Compared with si lncRNA FEZF1-AS1+OE vector group, the proliferation, invasion and migration of si lncRNA FEZF1-AS1+OE EZH2 group were increased (P＜0.05). E-cadherin expression was decreased, while N-cadherin, vimentin and EZH2 expressions were increased (P＜0.05). RIP experiment further confirmed that lncRNA FEZF1-AS1 had direct binding effect with EZH2. Conclusions LncRNA FEZF1-AS1 can promote the proliferation, invasion, metastasis and EMT process of pulmonary fibrosis cells by regulating EZH2.

Key words: idiopathic pulmonary interstitial fibrosis, FEZ family zinc finger 1 antisense RNA 1(FEZF1-AS1), epithelial-mesenchymal transition(EMT), enhancer of zeste homolog 2(EZH2), human lung adenocarcinoma cell line A549

中图分类号:

R563.9

王春燕, 王萍, 宋龙飞, 刘永全, 满君. LncRNA FEZF1-AS1通过调控EZH2对肺间质细胞增殖、迁移及侵袭的作用[J]. 基础医学与临床, 2024, 44(1): 43-50.

WANG Chunyan, WANG Ping, SONG Longfei, LIU Yongquan, MAN Jun. Effects of lncRNA FEZF1-AS1 on proliferation, migration and invasion through regulating EZH2 of lung interstitial cells[J]. Basic & Clinical Medicine, 2024, 44(1): 43-50.

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LncRNA FEZF1-AS1通过调控EZH2对肺间质细胞增殖、迁移及侵袭的作用

Effects of lncRNA FEZF1-AS1 on proliferation, migration and invasion through regulating EZH2 of lung interstitial cells

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