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Published on: September 3, 2021
Characterization of hepatocarcinoma cells: lymphatic metastasis potential through enhanced production, expression,
Jingwen Wang1, Xuejiao Qian2, Zhiqi Yin1
1Department of Pathology, Tianjin First Central Hospital, School of Medicine, Nankai University, No.2 Baoshan Road, Xiqing District, Tianjin, 300192, China.
Objectives:
In vitro and in vivo analyses were conducted to examine the correlations between the production, expression, and secretion of the VEGF-C/D-VEGFR-3/NRP-2 axis and various levels of lymphatic metastatic potential in murine hepatocellular carcinoma cells.
Methods:
Quantitative real-time PCR, western blotting, cytoimmunofluorescence, immunohistochemistry, and enzyme-linked immunosorbent assay were employed to assess the expression and secretion of the VEGF-C/D-VEGFR-3/NRP-2 axis at the gene, protein, cytological, and histological levels.
Results:
In both in vitro and in vivo experiments, the ligands VEGF-C/D and receptors VEGFR-3/NRP-2 were primarily localized in the cytoplasm or cell membrane of hepatocarcinoma F/P cells with high/low lymphatic metastatic potentials, and in normal hepatocytes, respectively. The production and expression levels of the VEGF-C/D-VEGFR-3/NRP-2 axis in F/P cells were significantly higher than those in normal liver cells. Additionally, the production and expression of this axis were higher in F cells compared to P cells (P < 0.01). Notably, within both F/P cells, the production and expression levels of VEGF-C and VEGFR-3 were lower than those of VEGF-D and NRP-2 (P < 0.05). However, the secretion of VEGF-C exceeded that of VEGF-D in the supernatant of cultured cells and in the serum of tumor-bearing mice (P < 0.01). Interestingly, the ratio of VEGF-C/D in F cells was significantly higher than that in P cells and normal hepatocytes. Moreover, the ratio of VEGF-C/D was notably elevated in the supernatant of tumor cells and serum from tumor-bearing mice, as determined by ELISA, compared to that in tumor cells and tissues assessed using other technologies (P < 0.01).
Conclusion:
The significant co-production, co-expression, and co-secretion of the VEGF-C/D-VEGFR-3/NRP-2 axis are distinctive features of hepatocarcinoma cells, particularly those with a heightened potential for lymphatic metastasis.
