Chu Changbin, Liu Shangjing, He Zhiting, Wu Mingjun, Xia Jing, Zeng Hongxiang, Xie Wenhua, Cheng Rui, Zhao Xueya, Li Xi
Institute of Life Sciences, Chongqing Medical University, Chongqing, 400016, China; Department of Urology, Chongqing Red Cross Hospital (People's Hospital of Jiangbei District), Chongqing, 400020, China.
School of Basic Medicine, Chongqing Medical University, Chongqing, 400016, China.
Transl Oncol. 2024 Nov;49:102112. doi: 10.1016/j.tranon.2024.102112. Epub 2024 Sep 2.
Clear cell renal cell carcinoma (ccRCC) is a serious threat to human life. It is very important to clarify the pathogenesis of ccRCC. In this study we evaluated the clinical value of HADH and explored its role and mechanism in the malignant progression of ccRCC.
HADH expression and its relationship with prognosis were analyzed using bioinformatics database. RT-PCR, Western blot and immunohistochemistry were used to examine the expression of HADH in ccRCC tissues and tissue microarrays. To examine the cell proliferation, apoptosis, migration and invasion ability, ccRCC cells with HADH overexpressed were constructed. Xenograft experiments were performed to determine the role of HADH. Non-target metabolomics was applied to explore the potential metabolic pathway by which HADH inhibited ccRCC progression. Plasmid pcDNA3.1-NRF2 was used to confirm whether HADH inhibited the process of ccRCC cells through NRF2-related glutathione (GSH) synthesis.
Bioinformatics database analysis showed that HADH expression was significantly decreased in ccRCC tissues, and its low expression predicted a poor prognosis. Both ccRCC tissues and tissue microarrays exhibited a significantly decreased HADH level compared with adjacent normal renal tissues. HADH overexpression inhibited the malignant behaviors of ccRCC cells. Furthermore, HADH overexpression attenuated GSH synthesis and induced oxidative stress damage. Exogenously increased NRF2 effectively attenuated the inhibitive effect of HADH overexpression on ccRCC cells.
Our data revealed that HADH suppressed the malignant behaviors of ccRCC cells by attenuating GSH synthesis through inhibition of NRF2 nuclear translocation, and HADH might be a novel therapeutic target for ccRCC treatment.
透明细胞肾细胞癌（ccRCC）对人类生命构成严重威胁。阐明ccRCC的发病机制非常重要。在本研究中，我们评估了HADH的临床价值，并探讨了其在ccRCC恶性进展中的作用及机制。
使用生物信息学数据库分析HADH表达及其与预后的关系。采用RT-PCR、蛋白质免疫印迹法和免疫组织化学检测ccRCC组织及组织芯片中HADH的表达。构建过表达HADH的ccRCC细胞，检测细胞增殖、凋亡、迁移和侵袭能力。进行异种移植实验以确定HADH的作用。应用非靶向代谢组学探索HADH抑制ccRCC进展的潜在代谢途径。使用质粒pcDNA3.1-NRF2确认HADH是否通过NRF2相关的谷胱甘肽（GSH）合成抑制ccRCC细胞的进程。
生物信息学数据库分析显示，ccRCC组织中HADH表达显著降低，其低表达预示预后不良。与相邻正常肾组织相比，ccRCC组织及组织芯片中HADH水平均显著降低。HADH过表达抑制ccRCC细胞的恶性行为。此外，HADH过表达减弱GSH合成并诱导氧化应激损伤。外源性增加NRF2可有效减弱HADH过表达对ccRCC细胞的抑制作用。
我们的数据表明，HADH通过抑制NRF2核转位减弱GSH合成，从而抑制ccRCC细胞的恶性行为，HADH可能是ccRCC治疗的新靶点。
