LIMCH1 通过与 HUWE1 相互作用抑制肺癌的生长，从而维持 p53 的稳定性。
LIMCH1 suppress the growth of lung cancer by interacting with HUWE1 to sustain p53 stability.
机构信息
Department of Laboratory Medicine, The Affiliated Huaian No.1 People's Hospital of Nanjing, Medical University, Huai'an, Jiangsu, 223300, China.
Department of Laboratory Medicine, The Affiliated Huaian No.1 People's Hospital of Nanjing, Medical University, Huai'an, Jiangsu, 223300, China.
出版信息
Gene. 2019 Sep 5;712:143963. doi: 10.1016/j.gene.2019.143963. Epub 2019 Jul 5.
BACKGROUND
The aim of this study was to identify the expression of LIM and calponin-homology domains 1 (LIMCH1) in lung cancer and normal tissues, to determine the interaction between LIMCH1 and HUWE1 in regulating p53 stability.
METHODS
The expression of LIMCH1 was detected by the Oncomine and Cancer Genome Atlas databases. Expression of LIMCH1 mRNA was identified using qRT-PCR. In transfected human lung cancer cells, co-immunoprecipitation experiments were performed. The mechanism that HUWE1 sustained lung cancer malignancy was verified by western blotting. The proliferation of tranfected cells was assessed by CCK-8 assay and colony formation.
RESULTS
Bioinformatic data and e TCGA database suggested LIMCH1 mRNA levels in tumor tissues were down-regulated compared to tumor adjacent tissues. We found low expression of LIMCH1 mRNA in tumor sites and tumor cell line. Exogenous expression of LIMCH1 interacts with HUWE1 promotes expression of p53. Use of siRNA or shRNA against LIMCH1 resulted in decreased p53 protein levels. LIMCH1 deletion lead to enhance of p53 ubiquitination and protein expression of p53 and substrate p21, puma. Growth curve showed that LIMCH1 deletion significantly promoted the proliferation of A549 cells.
CONCLUSIONS
LIMCH1 was a negative regulator and indicated a new molecular mechanism for the pathogenesis of lung cancer via modulating HUWE1 and p53.
背景
本研究旨在鉴定 LIM 和钙调蛋白同源结构域 1（LIMCH1）在肺癌和正常组织中的表达，确定 LIMCH1 与 HUWE1 之间在调节 p53 稳定性方面的相互作用。
方法
使用 Oncomine 和癌症基因组图谱数据库检测 LIMCH1 的表达。使用 qRT-PCR 鉴定 LIMCH1 mRNA 的表达。在转染的人肺癌细胞中进行共免疫沉淀实验。通过 Western blot 验证 HUWE1 维持肺癌恶性的机制。通过 CCK-8 测定和集落形成评估转染细胞的增殖。
结果
生物信息学数据和 eTCGA 数据库表明肿瘤组织中的 LIMCH1 mRNA 水平与肿瘤相邻组织相比下调。我们发现肿瘤部位和肿瘤细胞系中 LIMCH1 mRNA 表达水平较低。外源性表达的 LIMCH1 与 HUWE1 相互作用可促进 p53 的表达。使用针对 LIMCH1 的 siRNA 或 shRNA 可导致 p53 蛋白水平降低。LIMCH1 缺失导致 p53 泛素化和 p53 及其底物 p21、puma 的蛋白表达增强。生长曲线表明 LIMCH1 缺失可显著促进 A549 细胞的增殖。
结论
LIMCH1 是一种负调节剂，通过调节 HUWE1 和 p53 为肺癌的发病机制提供了新的分子机制。
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