Adipose-derived Mesenchymal Stem Cells Enhance The Growth And Metastasis Of Ovarian Cancer By Increasing EZH2 Expression

Object: The aim of this study was to investigate the role of EZH2 in the promotion of ovarian cancer by ADSCs.Methods:1、Omentum tissues were harvested from three healthy adult female donors(age range:39-45,BMI ages:22.5-25.8),the potential capacity of ADSCS to differentiate into osteoblasts and adipocytes were assessed.The expression of stromal makers were analyzed by flow cytometry.ADSCS were cultured with DMEM/F12 cultured medium containing 10% FBS and the culture medium were collected.2、Human ovarian cells lines(SKOV3,ES2 and HO8910)were obtained from Academy of Sciences Shanghai Cell Bank and cultured with DMEM/F12 cultured medium containing 10% FBS,all cells were grown in humidified 5% CO2,37 ℃ in cubator.3、Ovarian cell lines were cultured with ADSCS-CM or unconditioned medium;PCR and western blot were used to detect the mRNA and protein expression of EZH2;scratch-wound assay and 24-well Transwell plates were used to detected the migration and invasion abilites of ovarian cells.4、The ovarian cell lines were transfected with EZH2 shRNA using Lipofectamine 2000;stable transfected cell lines were selected followed by isolation of the monoclone;we used the methods above to detected the proliferation、migration and invasion ability of transfected cell lines.5、we stablished an orthotopic mouse model of ovarian cancer and monitored tumor cell behaviour using an vivo imaging system.6、Statistical analysis All data were expressed as the mean ± standard deviation;The significance of observed differences was analysed with a two-tailed Student’s t-test or one-way analysis of variance.A value of P<0.05 was considered significant.Results:1、The stable transfected cell lines were stablished and selected followed by isolation of the monoclone.2、The mRNA and protein expression of EZH2 significantly increased in cells cultured with ADSCS-CM compared with cells cultured with unconditioned medium(p<0.01).The migration and invasion ability significantly increased in cells cultured with ADSCS-CM compared with cells cultured with unconditioned medium(p<0.001).3、The migration and invasion ability significantly decreased in cells even cultured with ADSCS-CM compared with untransfected cells(p<0.001).Conclusion:ADSCs could promote the proliferation、migration and invasion ability by increasing the expression of EZH2.