Study on platelet enhancement of angiogenesis in hepatocellular car-cinoma and intervention effect of Cinobufagin
10.12092/j.issn.1009-2501.2024.08.003
- VernacularTitle:血小板增强肝癌血管生成及华蟾素的干预作用研究
- Author:
Xiaohong WU
1
;
Shiyu CHEN
;
Min CHEN
;
Mingjun YE
;
Shujun CHEN
;
Yangpei WU
;
Baikun LI
;
Qinglin LI
Author Information
1. 安徽中医药大学中医学院,合肥 230012,安徽
- Keywords:
liver cancer;
angiogenesis;
platelets;
human umbilical vein endothelial cells;
cinobufagin
- From:
Chinese Journal of Clinical Pharmacology and Therapeutics
2024;29(8):861-869
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To investigate the effect of plate-let(PLT)on angiogenesis in hepatocellular carcino-ma and the intervention effect of Cinobufagin(CBG).METHODS:Firstly,we screened the suitable co-incubation ratio of PLT and hepatocellular carci-noma cells,prepared conditioned medium,and de-termined the half inhibitory concentration of Cinob-ufagin;then,we set up a control group(human um-bilical vein endothelial cells(EC)+conventional me-dium),a crosstalk group(EC+CM_HP(strip culture prepared by crosstalk of HUH7 and PLT)),and an in-tervention group(EC+CM_HP+CBG).The migration,tube-formation and sprouting capacity of EC and the level of vascular endothelial growth factor(VEGF)in co-cultured supernatant were evaluated by scratch assay,tube-formation assay,budding as-say and ELISA assay.Western blot was used to de-tect the expression of VEGFR2 and p-VEGFR2,and reverse verification was performed with inhibitors.A subcutaneous transplantation tumour model of hepatocellular carcinoma in nude mice was estab-lished,with Model group,Model+CBG group and Model+Apa group.The collagen expression of the transplantation tumour was observed by Masson staining,and the expression levels of vascular endo-thelial markers CD31 and CD34 were detected by immunofluorescence.RESULTS:When PLT:HUH7=200,the activity of HUH7 was the strongest,and the crosstalk between HUH7 and PLT significantly promoted the proliferation of EC(P＜0.01).Com-pared with Control group,the migration,tube-for-mation and budding ability of Crosstalk group were enhanced,and those of Intervention group were lower than those of Crosstalk group(P＜0.01).The expression level of VEGF in the supernatant of Crosstalk group was higher than that of Control group,while that of Intervention group was lower than that of Crosstalk group(P＜0.01).The expres-sion level of p-VEGFR2 protein in Crosstalk group was significantly higher than that of Control group,but the expression level of Intervention group was lower than that of Crosstalk group(P＜0.01).Large collagen fibre deposition was seen in the Model group,and CBG intervention significantly reduced collagen fibre deposition in the transplanted tu-mour tissues.CD31 and CD34 expression was pres-ent in the hepatocellular carcinoma transplanted tumour tissues in the Model group,and CBG inter-vention significantly reduced the expression of CD31 and CD34 in the liver cancer transplanted tu-mour tissues(P＜0.01).CONCLUSION:PLT enhances angiogenesis in hepatocellular carcinoma,and CBG may inhibit its tube-forming ability via the VEGF/VEGFR2 pathway.
