Lu Jun, Chen Zhan, Zhao Hu, Dong Huiyue, Zhu Ling, Zhang Yi, Wang Jie, Zhu Hehuan, Cui Qiang, Qi Chuang, Wang Shuiliang, Chen Shushang, Shao Jichun
Fujian Provincial Key Laboratory of Transplant Biology, Fuzhou General Clinical College, Fujian Medical University, Fuzhou, 350025, China.
Fujian Provincial Key Laboratory of Transplant Biology, Dongfang Hospital (900 Hospital of the Joint Logistics Team), Xiamen University, Fuzhou, 350025, China.
J Transl Med. 2020 Feb 24;18(1):101. doi: 10.1186/s12967-020-02268-1.
Clear cell renal cell carcinoma (ccRCC) is a malignancy characterized by metabolic reprogramming. ABAT and ALDH6A1 are metabolic enzymes. In this study, we aim to investigate the associations of ABAT and ALDH6A1 with the malignancy of ccRCC cells.
The gene expression levels of ABAT and ALDH6A1 in ccRCC were analyzed from gene expression microarray datasets and RNA sequencing data. Clinical information was analyzed from The Cancer Genome Atlas (TCGA) data. The distributions of ABAT and ALDH6A1 in ccRCC clinical tissues were screened by reverse transcription-quantitative polymerase chain reaction (RT-QPCR) and immunohistochemical assays. The effect of overexpression of ABAT or ALDH6A1 was measured by detecting the cell viability, migration ability, and the ratio of lactate and nicotinamide adenine dinucleotide phosphate (NADPH). Chromatin immunoprecipitation (ChIP) and luciferase reporter assays were carried out to investigate the transcript regulation of HNF4A in ABAT and ALDH6A1.
Remarkable downregulated ABAT and ALDH6A1 expression levels were observed in ccRCC patients and low expression of ABAT and ALDH6A1 was correlated with poor survival. Overexpression of ABAT or ALDH6A1 significantly attenuated cell proliferation and migration, and impaired lactate production. In ABAT increased ccRCC cells, the ratio of NADPH/NADP+ was reduced. Finally, we demonstrated that ABAT and ALDH6A1 were directly regulated by a tumor suppressor, HNF4A.
These observations identified HNF4A-regulated low-expressed ABAT and ALDH6A1 as promising diagnostic and prognostic biomarkers for ccRCC.
透明细胞肾细胞癌（ccRCC）是一种以代谢重编程为特征的恶性肿瘤。ABAT和ALDH6A1是代谢酶。在本研究中，我们旨在探讨ABAT和ALDH6A1与ccRCC细胞恶性程度的相关性。
从基因表达微阵列数据集和RNA测序数据中分析ccRCC中ABAT和ALDH6A1的基因表达水平。从癌症基因组图谱（TCGA）数据中分析临床信息。通过逆转录定量聚合酶链反应（RT-QPCR）和免疫组织化学分析筛选ABAT和ALDH6A1在ccRCC临床组织中的分布。通过检测细胞活力、迁移能力以及乳酸与烟酰胺腺嘌呤二核苷酸磷酸（NADPH）的比率来测定ABAT或ALDH6A1过表达的影响。进行染色质免疫沉淀（ChIP）和荧光素酶报告基因分析以研究HNF4A对ABAT和ALDH6A1的转录调控。
在ccRCC患者中观察到ABAT和ALDH6A1表达水平显著下调，且ABAT和ALDH6A1低表达与较差的生存率相关。ABAT或ALDH6A1过表达显著减弱细胞增殖和迁移，并损害乳酸生成。在ABAT过表达的ccRCC细胞中，NADPH/NADP +的比率降低。最后，我们证明ABAT和ALDH6A1直接受肿瘤抑制因子HNF4A调控。
这些观察结果表明，HNF4A调控的低表达ABAT和ALDH6A1有望成为ccRCC的诊断和预后生物标志物。
