安徽医科大学学报2024,Vol.59Issue(4) :600-604.DOI:10.19405/j.cnki.issn1000-1492.2024.04.007

lncSIL通过EZH2/P21/CDK6信号通路负向调控TGF-β1诱导的肺泡上皮细胞间质转化

The lncSIL molecule exerts a negative regulatory effect on the alveolar epithelial-mesenchymal transition induced by TGF-β1 through modulation of the EZH2/P21/CDK6 signaling pathway

张万方 王琳 潘鹏涛 李文昕 康瑞丽 朱子任 陈浩勤 方新宇 张星灿 张雨昕 姜依雯 李欣妍 袁本琪
安徽医科大学学报2024,Vol.59Issue(4) :600-604.DOI:10.19405/j.cnki.issn1000-1492.2024.04.007

lncSIL通过EZH2/P21/CDK6信号通路负向调控TGF-β1诱导的肺泡上皮细胞间质转化

The lncSIL molecule exerts a negative regulatory effect on the alveolar epithelial-mesenchymal transition induced by TGF-β1 through modulation of the EZH2/P21/CDK6 signaling pathway

张万方 1王琳 2潘鹏涛 3李文昕 2康瑞丽 1朱子任 1陈浩勤 2方新宇 2张星灿 2张雨昕 2姜依雯 2李欣妍 2袁本琪2
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作者信息

  • 1. 新乡学院生物工程学院生物技术教研室,新乡 453003
  • 2. 新乡学院国际教育学院生物技术教研室,新乡 453003
  • 3. 新乡学院医学院基础医学教研室,新乡 453003
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摘要

目的 研究在转化生长因子β1(TGF-β1)诱导肺泡上皮细胞间质转化(EMT)进程中lncSIL的作用及其相关信号通路.方法 采用 Western blot 法研究沉默 lncSIL 后对TGF-β1 诱导 EMT 进程中细胞标志蛋白 E-钙黏蛋白(E-cad)、α-平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原蛋白(Col Ⅰ)表达的影响;通过RNA pulldown分析lncSIL相互作用蛋白,并检测过表达或沉默lncSIL后对其靶基因组蛋白赖氨酸N-甲基转移酶(EZH2)以及下游因子P21 蛋白(P21)和细胞周期蛋白依赖性激酶 6(CDK6)表达的影响,并结合流式细胞术分析lncSIL对细胞周期进程的作用.结果 沉默lncSIL后,间质细胞标志蛋白α-SMA和Col I表达升高,肺泡上皮细胞标志蛋白E-cad表达下降;RNA pulldown实验结果显示EZH2 是与lncSIL 相互作用的靶蛋白,并且沉默 lncSIL 后EZH2 表达升高,其下游基因 P21 表达下调,CDK6 表达上调,同时S期细胞的数量显著升高;过表达lncSIL时,EZH2与CDK6 表达下调,P21 表达上调,同时S期细胞的数量明显降低.结论 lncSIL通过负向调控EZH2/P21/CDK6 信号通路抑制细胞周期进程进而抑制TGF-β1 诱导的肺泡上皮细胞向间质转化.

Abstract

Objective To investigate the role of lncSIL in transforming growth factor-β1(TGF-β1)-induced alveo-lar epithelial interstitial transformation(EMT)and its related signaling pathways.Methods Western blot was used to detect the effect of lncSIL silencing on the expression of E-cadherin(E-cad),alpha-smooth muscle actin(α-SMA)and Collagen I(Col I)in the process of EMT induced by TGF-β1.LncSIL interacting proteins were ana-lyzed by RNA pulldown.Western blot was used to detect the effect of overexpression or silencing of lncSIL on the expression of its target gene enhancer of zeste homolog 2(EZH2)and its downstream factors P21 and cyclin-de-pendent kinase 6(CDK6).Flow cytometry was used to analyze the effect of lncSIL on cell cycle progression.Re-sults After lncSIL silencing,the expression of α-SMA and Col I increased,the expression of E-cad decreased.RNA pulldown assay showed that EZH2 was the target protein that interacted with lncSIL,and the expression of EZH2 increased after silencing lncSIL,the expression of EZH2 downstream gene P21 decreased,CDK6 increased.Flow cytometry showed that the number of cells in S phase significantly increased.When lncSIL was overexpressed,the expression of EZH2 and CDK6 was down-regulated,the expression of P21 was up-regulated,and the number of S phase cells significantly decreased.Conclusion LncSIL inhibits TGF-β1-induced alveolar epithelial cell mesen-chymal transition by negatively regulating EZH2/P21/CDK6 signaling pathway to inhibit cell cycle progression.

关键词

lncSIL/长链非编码RNA/特发性肺纤维化/上皮细胞间质转化/转化生长因子β1/Zeste同源物增强子2/细胞标志蛋白

Key words

lncSIL/long noncoding RNA/idiopathic pulmonary fibrosis/epithelial-mesenchymal transition/transforming growth factor-β1/enhancer of zeste homolog 2/cell marker proteins

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基金项目

河南省高等学校重点科研项目(23B180011)

国家自然科学基金(81702074)

出版年

2024
安徽医科大学学报
安徽医科大学

安徽医科大学学报

CSTPCD北大核心
影响因子:1.095
ISSN:1000-1492
参考文献量14
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