Xiang Jin, Zhang Yuhong, Tuo Lin, Liu Rui, Gou Dongmei, Liang Li, Chen Chang, Xia Jie, Tang Ni, Wang Kai
Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), Institute for Viral Hepatitis, Department of Infectious Diseases, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, 400010, China.
The Center for Clinical Molecular Medical Detection, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, China.
Genes Dis. 2019 Apr 16;7(1):150-159. doi: 10.1016/j.gendis.2019.04.004. eCollection 2020 Mar.
Phosphoenolpyruvate carboxykinase 1 (PCK1), a step limiting enzyme of gluconeogenesis, is downregulated in hepatocellular carcinoma (HCC). Overexpression of PCK1 has been shown to suppress hepatoma cell growth, but the underlying mechanism remains unclear. We used recombinant adenovirus overexpressing PCK1 or GFP in Huh7 cells, and the differentially expressed genes (DEGs) were identified by RNA-Seq. 180 were upregulated by PCK1 overexpression, whereas 316 were downregulated. Pathway analysis illustrated that PCK1 was closely correlated with Wnt signaling pathway and TGF-beta signaling pathway. Hence, Wnt signaling pathway and its downstream component, FZD2, FZD6, FZD7 and β-catenin were confirmed by qRT-PCR and Western blot. In vivo we also observed that PCK1 had restrained tumor growth as a result of decreasing expression of β-catenin. Whole-transcriptomic profile analysis discovered that overexpression of PCK1 downregulates several oncogenic signaling pathways in HCC, providing potential therapeutic targets for improving HCC therapy.
磷酸烯醇式丙酮酸羧激酶1（PCK1）是糖异生的限速酶，在肝细胞癌（HCC）中表达下调。PCK1过表达已被证明可抑制肝癌细胞生长，但其潜在机制仍不清楚。我们在Huh7细胞中使用过表达PCK1或绿色荧光蛋白（GFP）的重组腺病毒，并通过RNA测序鉴定差异表达基因（DEG）。PCK1过表达使180个基因上调，而316个基因下调。通路分析表明，PCK1与Wnt信号通路和转化生长因子-β（TGF-β）信号通路密切相关。因此，通过实时定量逆转录-聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法（Western blot）证实了Wnt信号通路及其下游成分卷曲蛋白2（FZD2）、卷曲蛋白6（FZD6）、卷曲蛋白7（FZD7）和β-连环蛋白。在体内，我们还观察到PCK1由于β-连环蛋白表达降低而抑制了肿瘤生长。全转录组图谱分析发现，PCK1过表达下调了HCC中的几种致癌信号通路，为改善HCC治疗提供了潜在的治疗靶点。
