The Interaction Of HA/CD44,nanog And STAT3 In Ovarian Cancer And Their Influence On The Expression Of MDR1 Protein

Objective To investigate the effects of HA/CD44 and Nanog on STAT3 expression in ovarian cancer SKOV-3 cells,and their influence on MDR1 protein.Methods(1)Detected the expression of STAT3 protein in ovarian cancer SKOV-3cells.Cultured the ovarian cancer SKOV-3 cells,then added HA in the culture medium and cultured for 24 hours;added anti-CD44 antibody in the culture medium and cultured for 1hours,then added HA and cultured for 24 hours.Extracted the nuclear protein and detected the expression of STAT3 protein by western blot.(2)Detected the effects of Nanog on the expression of STAT3 m RNA in ovarian cancer SKOV-3 cells.Added HA in the culture medium and cultured for 24 hours;added anti-CD44 antibody in the culture medium and cultured for 1 hour,then added HA and cultured for 24 hours;added the si RNA transfection mixture of Nanog in the culture medium and cultured for 48 hours,then added HA and cultured for 24 hours;added the c DNA transfection mixture of Nanog in the culture medium and cultured for 48 hours,then added HA and cultured for24 hours.Collected cells and isolated total RNA,detected the level of STAT3 m RNA by quantitative PCR.(3)Detected the regulation of MDR1 protein in ovarian cancer SKOV-3cells.Added HA in the culture medium and cultured for 24 hours;added anti-CD44 antibody in the culture medium and cultured for 1 hour,then added HA and cultured for 24hours;added the si RNA transfection mixture of Nanog in the culture medium and cultured for 48 hours,then added HA and cultured for 24 hours;added the c DNA transfection mixture of Nanog in the culture medium and cultured for 48 hours,then added HA and cultured for 24 hours;added the si RNA transfection mixture of STAT3 in the culture medium and cultured for 48 hours,then added HA and cultured for 24 hours.Extracted total protein and detected the expression of MDR1 protein by western blot.Results The relative level of STAT3 protein of the anti-CD44 antibody group and the group only added HA were 0.8271±0.0129?1.2984±0.0274,and the control group was1.0659±0.0085.The expression of STAT3 protein in the nuclear of SKOV-3 only treated by HA was about 1.5 times that of the group treated by anti-CD44 antibody,and the differences had statistical significance(p<0.05).The relative expression level of STAT-3m RNA in the control group;the addition of HA group;added anti-CD44 antibody and thenadded HA group;added transfection of Nanog si RNA and then added HA group;added transfection of Nanog c DNA and then added HA group was 2.0183±0.0849 ?2.4111±0.3856?1.0938±0.2045?1.6624±0.1938?6.5091±0.7458.The difference was statistically significant(P<0.05).The relative level of MDR1 protein in the control group;the addition of HA group;added anti-CD44 antibody and then added HA group;added transfection of Nanog si RNA and then added HA group;added transfection of Nanog c DNA and then added HA group;added transfection of STAT3 si RNA and then added HA group was 0.6334±0.0074?0.8701±0.0241?0.4695±0.0331?0.2982±0.0151?1.1149±0.0498?0.1388±0.0224.And the differences between each group had statistically significant(P<0.05).Conclusions Upregulation of HA/CD44 and Nanog can enhance the expression and activation of STAT3 in ovarian cancer SKOV-3 cells,and they can regulate the expression of MDR1 protein in ovarian cancer cells.