ERO1A 通过HIF1A调控EGFR的转录表达进而促进肺腺癌的进展

肺癌是世界范围内的主要公共卫生问题，也是癌症患者死亡的主要原因。而肺癌最常见的亚型是非小细胞肺癌，约占肺癌总数的85%，其中以肺腺癌最为突出，约占所有肺癌的40%。相比其他组织学类型的肺癌，肺腺癌的预防和治疗已成为一个更大的难题。因此，阐明肺腺癌发生发展和转移相关的分子机制并确定新的靶点对于肺腺癌的诊断和治疗具有重要意义。本项目针对这一问题，利用大数据分析发现内质网氧化酶1A（ERO1A）在肺癌组织中高表达且与病人预后不良相关，并且ERO1A在肺腺癌方面的研究鲜有报道。因此，本项目致力于明确该基因在肺腺癌发展过程中的功能和机制，为肺腺癌的治疗和预防提供新的思路和方法。通过细胞增殖、克隆形成、Transwell侵袭和迁移实验，我们发现敲低ERO1A可抑制肺癌细胞增殖、克隆形成、迁移及侵袭能力。我们用siRNAs敲降ERO1A后，进行RNA-seq测序，测序结果揭示ERO1A可能调控多个癌症相关信号通路。我们对测序结果进行分析发现ERO1A调控缺氧诱导转录因子1A（HIF1A）和表皮生长因子受体（EGFR）等肺癌发生、发展及转移相关的重要通路和基因。通过WB实验我们发现敲降ERO1A后，在多个肺癌细胞系中HIF1A和EGFR的表达都会显著下降，而敲降HIF1A后，EGFR的表达显著降低。此外，我们还发现HIF1A蛋白在EGFR基因的启动子区和其它部位均有结合位点。最后，我们通过肿瘤异种移植实验发现敲低ERO1A可抑制小鼠皮下移植瘤的生长。以上结果提示ERO1A在肺腺癌中可能通过HIF1A-EGFR信号通路促进肺腺癌的增殖和转移，并可能作为肺腺癌的预后标记物和治疗靶点。

Lung cancer is a major public health problem worldwide and the leading cause of death among cancer patients. The most common subtype of lung cancer is non-small cell lung cancer, which accounts for about 85% of all lung cancers, with lung adenocarcinoma being the most prominent, accounting for about 40% of all lung cancers. Compared to other histologic kinds of lung cancer, lung adenocarcinoma has a more difficult time being prevented and treated. The diagnosis and treatment of lung adenocarcinoma therefore depend on unraveling the molecular pathways involved in the growth and spread of the disease and locating new targets. In this project, we address this issue and use big data analysis to discover that endoplasmic reticular oxidoreductase 1A (ERO1A) is highly expressed in lung cancer tissues and correlates with poor patient prognosis, and that ERO1A is hardly ever reported in lung adenocarcinoma. Therefore, this project is dedicated to clarifying the function and mechanism of this gene in the development of lung adenocarcinoma, so as to provide new ideas and methods for the treatment and prevention of lung adenocarcinoma. We discovered that knockdown of ERO1A decreased the proliferation, clone formation, migration, and invasion capabilities of lung cancer cells using Transwell invasion and migration experiments. We performed RNA-seq sequencing after knockdown of ERO1A with siRNAs, and the sequencing results revealed that ERO1A may regulate several cancer-related signaling pathways. Our examination of the sequencing data showed that ERO1A controls crucial pathways and genes involved in the initiation, growth, and metastasis of lung cancer, including EGFR and HIF1A. Using Western Blot experiment, we discovered that in several lung cancer cell lines, knocking down ERO1A led to a large decrease in the expression of hypoxia-inducible factor 1A (HIF1A) and epidermal growth factor receptor (EGFR), while knocking down HIF1A resulted in a considerable decrease in the expression of EGFR. We also found that the EGFR gene's promoter region and other regions contain binding sites for the HIF1A protein. Finally, using tumor xenograft experiments, we discovered that knockdown of ERO1A prevented the development of subcutaneous graft tumors in mice. The above results suggest that ERO1A in lung adenocarcinoma may promote the proliferation and metastasis of lung adenocarcinoma through the HIF1A-EGFR signaling pathway and may serve as a prognostic marker and therapeutic target for lung adenocarcinoma.

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