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VEGF/Nrp1/HIF-1α promotes proliferation of hepatocellular carcinoma through a positive feedback loop

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Abstract

To investigate the role of neuropilin1 (Nrp1) in glucose metabolism and proliferation of hepatocellular carcinoma (HCC) cells and to analyze its mechanism of action. The CRISPR gene knockout technique was used to knock out the Nrp1 gene in two HCC cell lines. The effect of Nrp1 on the proliferation of HCC cells was assessed in the CCK8 assay and plate cloning assay. The expression levels of glucose consumption, lactate production, and essential proteins of the glycolytic pathway were detected to explore the effect of Nrp1 on glucose metabolism in HCC cells. Using CoCl2 to revert the expression of hypoxia inducible factor-1α (HIF-1α), the role of HIF-1α in the pro-HCC cell metabolism of Nrp1 were demonstrated. The protein synthesis inhibitor CHX and proteasome inhibitor MG-132 was used to analyze the molecular mechanism of action of Nrp1 on HIF-1α. The Kaplan–Meier method was used to calculate survival rates and plot survival curves. Based on the CCK8 assay and plate cloning assay, we found that Nrp1 knockout significantly inhibited the proliferation of HCC cells. Nrp1 inhibitor suppressed lactate production and glucose consumption in HCC cells. Knockout of Nrp1 decreased the expression of glycolytic pathway-related proteins and HIF-1α protein. Furthermore, by joint use of CoCl2 and NRP1 knockout, we confirmed that reverting HIF-1α expression could reverse the effect of Nrp1 knockout on HCC cell metabolism in vitro. Mechanistically, Nrp1 showed a close correlation with the stability of HIF-1α protein in protein stability assay. Finally, we revealed that high expression of Nrp1 in HCC tissues was associated with poor overall survival and disease-free survival of the patients. Nrp1 accelerates glycolysis and promotes proliferation of HCC by regulating HIF-1α protein stability and through the VEGF/Nrp1/HIF-1α positive feedback loop.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

The study was funded by Chongqing medical scientific research project (Joint project of Chongqing Health Commission and Science and Technology Bureau: No. 2021MSXM139), Postdoctoral Science Foundation of Chongqing Natural Science Foundation (No. cstc2020jcyj-bshX0033), and General program of Chongqing Natural Science Foundation (No. cstc2021jcyj-msxmX0294).

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Authors and Affiliations

  1. Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Chongqing Medical University, No. 288, Chayuan Tianwen Avenue, Nan’an District, Chongqing, People’s Republic of China

    Yun-Bing Wang, Kai-Wen Zheng, Haitham Salameen, Zhe-Yu Zhu, Fei-Fan Wu & **ong Ding

  2. Department of Hepatobiliary Surgery, The People’s Hospital of Rongchang District, Chongqing, People’s Republic of China

    Kai-Wen Zheng

  3. The Second Clinical College, Chongqing Medical University, Chongqing, People’s Republic of China

    Yi-Yu Hu

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Contributions

Conceptualization and design: YBW, KWZ, ZYZ, and XD; Methodology: YBW, KWZ, YYH, SH, and XD; In vitro and in vivo experiment: YBW, KWZ, FFW, and SH; Writing-review and editing: YBW, KWZ, YYH, SH, ZYZ, and XD; Funding acquisition: YBW and XD; Supervision: XD. All authors have read and approved the final manuscript.

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Correspondence to **ong Ding.

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The authors have no relevant financial or nonfinancial interests to disclose.

Ethical approval

This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of the second affiliated hospital of Chongqing Medical University.

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Wang, YB., Zheng, KW., Hu, YY. et al. VEGF/Nrp1/HIF-1α promotes proliferation of hepatocellular carcinoma through a positive feedback loop. Med Oncol 40, 339 (2023). https://doi.org/10.1007/s12032-023-02202-7

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