Functional mechanism of vascular endothelial growth factor in promoting the growth of intrahepatic cholangiocarcinoma

doi:10.3877/cma.j.issn.1674-0793.2019.02.003

Abstract

Abstract:

Objective

To investigate the role of vascular endothelial growth factor (VEGF) and VEGF receptors on cell growth of intrahepatic cholangiocarcinoma (ICC).

Methods

Western blotting was performed to detect the expression of VEGF in tumour tissues and paired normal tissues from twelve patients with ICC. ICC cell line Huh28 was treated with exogenous recombinant human VEGF (rhVEGF). Cell growth was evaluated by cell counting, proliferation was detected by BrdU cell proliferation assay, and apoptosis was detected by flow cytometry. The expression of VEGF receptors VEGFR1/VEGFR2 in Huh28 cells after rhVEGF treatment were detected by Western blotting. VEGFR1 and VEGFR2 were blocked by specific antibodies, and cell apoptosis was examined by apoptosis-ELISA assay. The stably knockdown VEGFR2 cell line Huh28-shVEGFR2 (experimental group) and the control cell line Huh28-shNC (control group) were established using shRNA lentivirus. Subcutaneous tumour models in ten nude mice with Huh28-shVEGFR2 and Huh28-shNC were used to observe tumour growth.

Results

The protein expression of VEGF was up-regulated in ICC tumour tissues than in matched normal tissues (P<0.01). Exogenous rhVEFG could promote the growth of Huh28 cells, suppressed cell apoptosis, without significant effect on cell proliferation ability. The expression of phosphorylated VEGFR1 and VEGFR2 in Huh28 cells were up-regulated after rhVEGF treatment (P<0.05). VEGFR2 antibody could significantly reverse the anti-apoptosis effect of rhVEGF (P<0.05), while VEGFR1 antibody did not. Subcutaneous tumour models indicated that tumour growth in experimental group (Huh28-shVEGFR2) was significantly inhibited compared with the control group (Huh28-shNC), the difference was significant (P<0.05).

Conclusion

VEGF promotes ICC cells growth through inhibiting apoptosis of ICC cells in a VEGFR2-dependent signalling pathway.

Key words: Intrahepatic cholangiocarcinoma, VEGF, VEGFR2, Apoptosis

Yifei Wang, Ruiming Liang, Guangyan Wei, Hongpeng Chu. Functional mechanism of vascular endothelial growth factor in promoting the growth of intrahepatic cholangiocarcinoma[J]. Chinese Archives of General Surgery(Electronic Edition), 2019, 13(02): 97-102.

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References 19

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