Zhuang Wenyuan, Shi Xiaokai, Gao Shenglin, Qin Xihu
Department of Urology, The Affiliated Changzhou Second People's Hospital of Nanjing Medical University, Changzhou, 213003, China.
Gonghe County Hospital of Traditional Chinese Medicine, Hainan Prefecture, Qinghai Province, China.
Cancer Metab. 2023 Aug 8;11(1):11. doi: 10.1186/s40170-023-00312-4.
Kidney renal clear cell carcinoma (KIRC) is the major histological subtype of kidney tumor which covers approximately 80% of the cases. Although various therapies have been developed, the clinical outcome remains unsatisfactory. Metabolic dysregulation is a key feature of KIRC, which impacts progression and prognosis of the disease. Therefore, understanding of the metabolic changes in KIRC is of great significance in improving the treatment outcomes.
The glycolysis/gluconeogenesis genes were analyzed in the KIRC transcriptome from the Cancer Genome Atlas (TCGA) by the different expression genes (DEGs) test and survival analysis. The gluconeogenesis-related miRNAs were identified by ImmuLncRNA. The expression levels of indicated genes and miRNAs were validated in KIRC tumor and adjunct tissues by QPCR. The effects of miR-4477b and PCK1 on cell proliferation and apoptosis were examined using the cell viability assay, cell apoptosis assay, and clone information. The interaction of miR-4477b with TEF was tested by the luciferase report gene assay. The different gluconeogenesis statuses of tumor cells and related signatures were investigated by single-cell RNA sequencing (scRNA-seq) analysis.
The 11 gluconeogenesis genes were found to be suppressed in KIRC (referring as PGNGs), and the less suppression of PGNGs indicated better survival outcomes. Among the 11 PGNGs, we validated four rate-limiting enzyme genes in clinical tumor patients. Moreover, restoring gluconeogenesis by overexpressing PCK1 or TEF through miR-4477b inhibition significantly inhibited tumor cell proliferation, colony formation, and induced cell apoptosis in vitro. Independent single-cell RNA sequencing (scRNA-seq) data analysis revealed that the tumor cells had high levels of PGNG expression (PGNG + tumor cells) represented a phenotype of early stage of neoplasia and prompted immune surveillance.
Our study suggests that the deficiency of gluconeogenesis is a key metabolic feature of KIRC, and restoring gluconeogenesis could effectively inhibit the proliferation and progression of KIRC cells.
肾透明细胞癌（KIRC）是肾肿瘤的主要组织学亚型，约占病例的80%。尽管已开发出多种治疗方法，但临床结果仍不尽人意。代谢失调是KIRC的一个关键特征，它影响疾病的进展和预后。因此，了解KIRC中的代谢变化对改善治疗结果具有重要意义。
通过差异表达基因（DEG）检测和生存分析，对癌症基因组图谱（TCGA）中KIRC转录组的糖酵解/糖异生基因进行分析。通过ImmuLncRNA鉴定糖异生相关的miRNA。通过QPCR验证KIRC肿瘤组织和癌旁组织中指定基因和miRNA的表达水平。使用细胞活力测定、细胞凋亡测定和克隆形成实验检测miR-4477b和PCK1对细胞增殖和凋亡的影响。通过荧光素酶报告基因实验检测miR-4477b与TEF的相互作用。通过单细胞RNA测序（scRNA-seq）分析研究肿瘤细胞的不同糖异生状态和相关特征。
发现11个糖异生基因在KIRC中受到抑制（称为PGNGs），PGNGs抑制程度越低，生存结果越好。在这11个PGNGs中，我们在临床肿瘤患者中验证了4个限速酶基因。此外，通过抑制miR-4477b过表达PCK1或TEF来恢复糖异生，可显著抑制体外肿瘤细胞增殖、集落形成并诱导细胞凋亡。独立的单细胞RNA测序（scRNA-seq）数据分析显示，具有高水平PGNG表达的肿瘤细胞（PGNG+肿瘤细胞）代表肿瘤形成早期的一种表型，并促进免疫监视。
我们的研究表明，糖异生缺陷是KIRC的关键代谢特征，恢复糖异生可有效抑制KIRC细胞的增殖和进展。
