Abstract
Objective:To investigate the effects of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) on apoptosis and proliferation of human ovarian cancer SKOV3 cells and to explore mechanism.Methods:hUC-MSCs were isolated and cultured by tissue block adherent culture method. The hUC-MSCs phenotype were identified by flow cytometry. The hUC-MSCs lysate and conditioned medium,directly combine were used to treat SKOV3 cells.The effects on the proliferation，apoptosis，mechanism of SKOV3 cells were examined by cell counting kit-8(CCK-8)，Annexin V-FITC/PI，quantitative real time polymerase chain reaction(RT-qPCR) and spheroid formation assays.Establish ovarian cancer xenograft models，1X 106 hUC-MSCs and 2 X 106 hUC-MSCs were administrated into the mice t rear back tumor tissue. After three injections of hUC-MSCs, the nude mice were sacrificed after 1 week of observation.Remove tumor tissue. Observed tumor volume changes every 3 days after the start of the experiment. The expression of CD34 and VEGF were detected by immunohistochemistry.Results:Human umbilical cord mesenchymal stem cells were cultured and isolated from tissue block. Flow cytometry results revealed that the hUC-MSCs marks CD44 and CD29, but not CD45 and CD34 were expressed on obtained cells. The apoptosis of SKOV3 cells was induced by hUC-MSCs lysate, conditioned medium and Transwell co-culture method in SKOV3 cells, and the apoptosis rate was higher with increasing concentration. hUC-MSCs conditioned medium and Transwell co-culture method can inhibit cell proliferation. After adding experimental factors, the conditioned medium and Transwell co-culture method can down-regulate the transcription of PI3KCA, AKT and BCL-2 genes in SKOV3 cells, and up-regulate the Caspase-3 gene.The tumor volume of the experimental group was smaller than that of the control group during the observation period. The expression levels of CD34 and VEGF in the experimental group were significantly lower than those in the control group(P<0.05).Conclusion: The conditioned medium of hUC-MSCs and the co-culture method of hUC-MSCs and SKOV3 can significantly inhibit the proliferation of SKOV3 cells, which is mainly achieved by inhibiting PI3K/AKT signaling pathway. hUC-MSCs can inhibit the growth of subcutaneous subcutaneous transplantation and the expression of CD34 and VEGF in ovarian cancer. It provides a new idea for the treatment of ovarian cancer.
Gene therapy of ovarian cancer using IL-21-secreting human umbilical cord mesenchymal stem cells in nude mice