摘要
目的:探讨长链非编码RNA LINC00472在肾纤维化细胞模型中的表达及其在TGF-β1诱导的肾小管上皮细胞纤维化和上皮间充质转化(EMT)中的功能作用。方法:使用重组人TGF-β1诱导人肾小管上皮细胞(HK-2)纤维化和EMT,采用实时荧光定量PCR法检测细胞中纤维化相关基因和LINC00472的mRNA表达水平,通过Western blot免疫印迹法检测细胞中纤维化相关基因的蛋白表达水平,通过划痕实验评估LINC00472表达对HK-2细胞迁移能力的影响。结果:TGF-β1能成功诱导HK-2细胞发生纤维化和EMT,并剂量和时间依赖性地抑制LINC00472的表达(P<0.05)。抑制LINC00472进一步促进TGF-β1诱导的Fibronectin和Vimentin上调,以及E-cadherin下调(P<0.05);而过表达LINC00472则能逆转TGF-β1对纤维化相关基因的诱导作用(P<0.05)。此外,抑制LINC00472能进一步增强TGF-β1诱导的HK-2细胞迁移(P<0.05),而上调LINC00472则使细胞迁移受到抑制(P<0.05)。结论:LINC00472在TGF-β1诱导的肾纤维化细胞模型中呈低表达,其表达水平的降低能促进细胞纤维化和EMT过程。
Objective: To investigate the expression of long non-coding RNA LINC00472 in an in vitro model of renal fibrosis and functional roles in renal fibrosis and tubular epithelial-mesenchymal transition(EMT). Methods: Recombinant human TGF-β1 were used to induce fibrosis and EMT of human renal tubular epithelial cells(HK-2). The m RNA expression levels of fibrosis-related genes and LINC00472 were measured by quantitative real-time PCR analysis, and the protein expression levels of fibrosis-related genes were detected by Western blot analysis. Wound healing assay was carried out to evaluate the impact of LINC00472 expression on HK-2 cells migration. Results: TGF-β1 successfully induced fibrosis and EMT of HK-2 cells, and inhibited LINC00472 expression in a dose-and time-dependent manner(P<0.05). LINC00472 knockdown further enhanced TGF-β1-induced upregulation of Fibronectin and Vimentin,and downregulation of E-cadherin(P<0.05), whereas LINC00472 overexpression significantly reversed the induction of fibrosis-related genes by TGF-β1(P<0.05). Furthermore, LINC00472 inhibition further promoted TGF-β1-induced cell migration(P<0.05), while LINC00472 upregulation hindered cell migration(P<0.05). Conclusions: LINC00472 is low-expressed in TGF-β1-induced in vitro model of renal fibrosis, and its downregulation aggravates fibrosis and EMT of renal tubular cells.
作者
徐超亮
陈磊
李登
陈飞腾
徐子杰
邵怡
沙明磊
XU Chao-liang;CHEN Lei;LI Deng;CHEN Fei-teng;XU Zi-jie;SHAO Yi;SHA Ming-lei(Department of Urology,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai,200080,China;Department of Geriatric,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai,200080,China)
出处
《现代生物医学进展》
CAS
2021年第11期2007-2011,共5页
Progress in Modern Biomedicine
基金
上海市浦江人才计划项目(2020PJD046)
上海市卫生健康委员会科研项目(202040071)。
关键词
肾纤维化
上皮间充质转化
慢性肾脏病
长链非编码RNA
Renal fibrosis
Epithelial-mesenchymal transition
Chronic kidney disease
Long non-coding RNAs
