The Effects of Grape Seed Proanthocyanidins on Human Osteosarcoma 143B Cell Line and Its Mechanisms

Abstract: To investigate the effect of grape seed proanthocyanidins (GSP) on cell proliferation and apoptosis， and its relevant mechanisms in human osteosarcoma cells， human osteosarcoma 143B cell line was treated with different concentration of GSP. The cell proliferation activity was detected by crystal violet staining and colony forming assay. The cell cycle and apoptosis of 143B cells were determined by flow cytometry. Overexpressing β-catenin by infection of adenovirus Adβ-catenin was performed to investigate whether Wnt/ β-catenin signaling pathway was involved in the GSP-induced proliferation suppression in 143B cells. RT-PCR and Western blot assay were used to detect the mRNA level and protein level of the related proteins， respectively. The result of crystal violet staining showed that 20~60 μg/mL GSP inhibited the cell proliferation activity in a dose- and time-dependent manner. The colony forming assay showed that GSP treatment decreased the number of colony. GSP induced G0/G1 phase arrest by the flow cytometry analysis and decreased the cell cycle related protein cyclin D1. The apoptosis rate and the apoptosis related proteins cleaved PARP (poly ADP-ribose polymerase) and cleaved caspase-8 were higher than their controls. At the same time， GSP increased GSK3β protein and reduced β-catenin mRNA， p-GSK3β， total β-catenin protein， nuclear β-catenin protein， and cyclin D1 protein. β-catenin overexpression partially attenuated the inhibition effect of GSP on cell proliferation. Ourstudy demostrated that GSP could inhibit the proliferation and promote the apoptosis in human osteosarcoma 143B cells， and its mechanism of the proliferation inhibition supposed to be related to inhibition of Wnt/β-catenin signaling pathway.

CSTR: 32200.14.cjcb.2017.12.0005