YANG Ying, LIU Ji-cheng, CONG Huan, et al. Effect and Related Machanism of 12-deoxyphorbol-13-palmitate on Expression of VEGF in Breast Cancer MCF-7 Cells[J]. Chinese journal of experimental traditional medical formulae, 2015, 21(2): 120-124.
YANG Ying, LIU Ji-cheng, CONG Huan, et al. Effect and Related Machanism of 12-deoxyphorbol-13-palmitate on Expression of VEGF in Breast Cancer MCF-7 Cells[J]. Chinese journal of experimental traditional medical formulae, 2015, 21(2): 120-124. DOI: 10.13422/j.cnki.syfjx.2015020120.
Objective: To observe the effect of 12-deoxyphorbol-13-palmitate (DP) which isolated from the roots of Euphorbia fischeriana on the vascular endothelial growth factor (VEGF) expression
and whether it shows anti-VEGF effects via Von Hippel-Lindau(VHL)/hypoxia induced 1 alphal(HIF-1α) signaling pathway. Method: The cells were divided into normoxia control group
normoxia experimental groups (10
20
40 μmol · L-1 DP)
hypoxia control group
hypoxia experimental groups (10
20
40 μmol · L-1 DP). The hypoxia control group and experimental groups were pretreated with CoCl2(150 μmol · L-1). The viability of MCF-7 cells was measured by MTT method at a cell density of 70%-80% and at 6
12
24 h. The secretion of VEGF was investigated by enzyme-linked irnmuno sorbent assay(ELISA).The mRNA level of VEGF was detected by RT-qPCR. The protein expressions of VEGF
HIF-1α and VHL were determined by Western blot. The localization and expression of HIF-1α were detected by immunofluorescence. Result: MTT assay showed that DP significantly decreased the cell viability at 12
24 h (P<0.05) and at DP concentration of 20 μmol · L-1 and 40 μmol · L-1. Under hypoxia condition
the expression of VEGF and HIF-1α was increased compared with that under normoxia condition. But the up-regulation was reversed (P<0.05) by the addition of DP. The VHL level was down-regulated under hypoxia condition. However after treatment of DP
the expression of VHL was increasing significantly (P<0.05). Conclusion: DP could down-regulate VEGF by affecting VHL/HIF-1α signaling pathway.