Wnt5a modulates vincristine resistance through PI3K/Akt/GSK3β signaling pathway in human ovarian carcinoma SKOV3/VCR cells.
- Author:
Feng-Lan WU
1
;
Hong-Lian CHEN
1
;
Xiao-Wei HU
1
;
Li-Ying LIANG
1
;
Wan-Ling XU
2
Author Information
1. Department of Medicine, Luohe Medical College, Luohe 462002, China.
2. Department of Medicine, Luohe Medical College, Luohe 462002, China. 872189991@qq.com.
- Publication Type:Journal Article
- MeSH:
ATP Binding Cassette Transporter, Subfamily B;
metabolism;
Cell Line, Tumor;
Drug Resistance, Neoplasm;
Female;
Gene Silencing;
Glycogen Synthase Kinase 3 beta;
metabolism;
Humans;
Ovarian Neoplasms;
pathology;
Phosphatidylinositol 3-Kinases;
metabolism;
Proto-Oncogene Proteins c-akt;
metabolism;
Signal Transduction;
Survivin;
metabolism;
Vincristine;
pharmacology;
Wnt-5a Protein;
metabolism
- From:
Acta Physiologica Sinica
2019;71(3):415-423
- CountryChina
- Language:Chinese
-
Abstract:
The aim of this study was to investigate the effect of Wnt5a on the vincristine (VCR) resistance in human ovarian carcinoma SKOV3 cells and its possible mechanism. The drug-resistant SKOV3/VCR cells were established by stepwise exposure to VCR, and then the SKOV3/VCR cells were stably transfected with specific shRNA interference plasmid vector targeting for Wnt5a. The mRNA expression level of Wnt5a was measured by RT-PCR. CCK-8 assay was used to detect the cell viability of SKOV3/VCR cells. The apoptosis was analyzed by flow cytometry. The protein expression levels of Wnt5a, MDR1, Survivin, β-catenin, Akt, p-Akt(S473), GSK3β and p-GSK3β(Ser9) were detected by Western blot. The result showed that SKOV3/VCR cells had significantly higher protein expression levels of Wnt5a, MDR1, Survivin and β-catenin, phosphorylation levels of Akt and GSK3β, and mRNA expression level of Wnt5a, compared with SKOV3 cells (P < 0.05). WNT5A gene silencing significantly increased the sensitivity of SKOV3/VCR cells to VCR, the IC of VCR being decreased from 38.412 to 9.283 mg/L (P < 0.05), synergistically enhanced VCR-induced apoptosis of SKOV3/VCR cells (P < 0.05), down-regulated the protein expression levels of MDR1, β-catenin and Survivin (P < 0.05), and inhibited phosphorylation of Akt and GSK3β (P < 0.05). Meanwhile, LY294002 (PI3K inhibitor) decreased the protein expression levels of MDR1, β-catenin and Survivin, as well as the phosphorylation levels of Akt and GSK3β in SKOV3/VCR cells (P < 0.05). These results suggest that WNT5A gene silencing reverses VCR resistance in SKOV3/VCR cells possibly through blocking the PI3K/Akt/GSK3β/β-catenin signaling pathway, and thus down-regulating the protein expression levels of MDR1 and Survivin.
