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RNA-seq Analysis of Transcriptomes in Thrombin-treated and Control Human Pulmonary Microvascular Endothelial Cells
Published on: February 13, 2013
Transcriptomic changes associated with PCK1 overexpression in hepatocellular carcinoma cells detected by RNA-seq
Jin Xiang1, Yuhong Zhang2, Lin Tuo1
1Key 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.
Abstract:
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.
Insights
Overexpressing Phosphoenolpyruvate carboxykinase 1 (PCK1) in liver cancer cells suppresses growth by downregulating oncogenic pathways like Wnt signaling. This reveals PCK1 as a potential therapeutic target for hepatocellular carcinoma (HCC).
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Phosphoenolpyruvate carboxykinase 1 (PCK1) is crucial for gluconeogenesis and is downregulated in hepatocellular carcinoma (HCC).
- PCK1 overexpression inhibits hepatoma cell growth, but the mechanism is not fully understood.
Purpose of the Study:
- To elucidate the molecular mechanisms by which PCK1 suppresses HCC cell growth.
- To identify signaling pathways affected by PCK1 overexpression in HCC.
Main Methods:
- Recombinant adenovirus vectors were used to overexpress PCK1 in Huh7 HCC cells.
- RNA-sequencing (RNA-Seq) identified differentially expressed genes (DEGs).
- Quantitative reverse transcription PCR (qRT-PCR) and Western blot validated gene and protein expression changes.
Main Results:
- PCK1 overexpression altered the expression of 180 upregulated and 316 downregulated genes.
- Pathway analysis revealed correlations between PCK1 and Wnt/TGF-beta signaling pathways.
- PCK1 overexpression decreased the expression of beta-catenin, a key Wnt pathway component, and inhibited tumor growth in vivo.
Conclusions:
- PCK1 overexpression downregulates oncogenic signaling pathways, including Wnt signaling, in HCC.
- PCK1 acts as a tumor suppressor in HCC by inhibiting beta-catenin expression.
- Targeting PCK1 may offer a novel therapeutic strategy for HCC treatment.
