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. 2022 Sep 23:12:998295.
doi: 10.3389/fonc.2022.998295. eCollection 2022.

HIF-2α regulates proliferation, invasion, and metastasis of hepatocellular carcinoma cells via VEGF/Notch1 signaling axis after insufficient radiofrequency ablation

Yongguang Yang  1   2 , Weifeng Chen  2 , Weiheng Mai  2 , Yi Gao  1   3
Affiliations

HIF-2α regulates proliferation, invasion, and metastasis of hepatocellular carcinoma cells via VEGF/Notch1 signaling axis after insufficient radiofrequency ablation

Yongguang Yang et al. Front Oncol. .

Abstract

Background and aims: Although insufficient radiofrequency ablation (RFA) promotes the recurrence and metastasis of liver cancer, the underlying mechanism remains unclear. This study aimed to investigate the role and mechanism of HIF-2α in hepatocellular carcinoma cells (HCCs) after Insufficient RFA.

Methods: We established a model of insufficient RFA in MHCC97H hepatoma cells and screened for stable sublines. We inhibited HIF-2α expression in the Insufficient RFA group using PT2385 and assessed the resulting changes in proliferation and biological function of HCCs. Cell viability and proliferation were detected by the MTT method, and scratch and Transwell chamber invasion tests detected migration and invasion abilities of HCCs. The mRNA and protein expression levels of VEGF, HIF-2α, and Notch1 were detected using qPCR, immunofluorescence, and western blotting.

Results: Compared with normal HCCs without RFA treatment, insufficient RFA enhanced the proliferation and invasion abilities of hepatocellular carcinoma subline MHCC97H (P < 0.001), as well as their migration ability (P = 0.046). The HIF-2α-specific inhibitor PT2385 downregulated the migration (P = 0.009) and invasion (P < 0.001) of MHCC97H cells but did not affect cell proliferation (P > 0.05). Insufficient ablation increased the mRNA and protein expression of VEGF, HIF-2α, and Notch1 in HCCs, whereas inhibition of HIF-2α reversed these changes.

Conclusions: Insufficient RFA increases the proliferation, migration, and invasion of HCCs via the HIF-2α/VEGF/Notch1 signaling axis; HIF-2α is a potential target for novel treatments of HCC after insufficient RFA.

Keywords: hepatocellular carcinoma; hypoxia-inducible factor-2α; metastasis; radiofrequency ablation; residual carcinoma.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Expression of HIF-2α, Notch1, and VEGF in HC and their paracancerous tissues of patients after insufficient RFA as revealed by immunohistochemistry. Brown color indicates positive staining. Magnification 400×.
Figure 2
Figure 2
HIF-2α, Notch1, and VEGF were involved in the increased invasion and proliferation of HCC induced by insufficient RFA. (A) Representative photos of HCC after insufficient RFA were detected by a phase contrast microscope and crystal violet staining. Magnification 400×. (B) Detection of cell proliferation by MTT. (C) Quantitation analysis of Transwell assay. (D–F) Detection of the mRNA expression of HIF-2α, Notch1, and VEGF. **P < 0.01, ***P < 0.001.
Figure 3
Figure 3
Inhibition of HIF-2α by PT2385 suppressed the invasion and migration of HCCs after insufficient RFA. (A) Representative photos of HCC after insufficient RFA were detected by wound healing assay. Magnification 200×. (B) Quantitation analysis of wound healing assay. (C, D) Representative photos and quantitation analysis of Transwell assay. Magnification 400×. *P < 0.05; **P < 0.01. ***P < 0.001.
Figure 4
Figure 4
Inhibition of HIF-2α by PT2385 suppressed the VEGF and Notch1 signaling pathway in HCC after insufficient RFA. (A–C) Detection of mRNA expression by RT-qPCR. (D, E) Detection of protein levels by western blotting. *P < 0.05, **P < 0.01, ***P < 0.001.
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References

    1. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. . Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin (2021) 71(3):209–49. doi: 10.3322/caac.21660 - DOI - PubMed
    1. Yang WS, Zeng XF, Liu ZN, Zhao QH, Tan YT, Gao J, et al. . Diet and liver cancer risk: a narrative review of epidemiological evidence. Br J Nutr (2020) 124(3):330–40. doi: 10.1017/S0007114520001208 - DOI - PubMed
    1. Lee SK, Lee SW, Jang JW, Bae SH, Choi JY, Yoon SK. Immunological markers, prognostic factors and challenges following curative treatments for hepatocellular carcinoma. Int J Mol Sci (2021) 22(19):10271. doi: 10.3390/ijms221910271 - DOI - PMC - PubMed
    1. Rossi S, Fornari F, Pathies C, Buscarini L. Thermal lesions induced by 480 KHz localized current field in guinea pig and pig liver. Tumori (1990) 76(1):54–7. doi: 10.1089/sct.1990.6.51 - DOI - PubMed
    1. Zervas NT, Kuwayama A. Pathological characteristics of experimental thermal lesions. comparison of induction heating and radiofrequency electrocoagulation. J Neurosurg (1972) 37(4):418–22. doi: 10.3171/jns.1972.37.4.0418 - DOI - PubMed

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