Effects Of Nanog Gene And STAT3 On The Growth Of Ovarian Cancer Cells In HA And CD44 Signaling Pathway

Objective:To detect the expression and interaction of Nanog and STAT3 in epithelial ovarian cancer,to investigate the interaction in HA?CD44 signal pathway,synergistic effect of Nanog gene and STAT3 and their influence on the growth of ovarian cancer cells,and provide scientific evidence for the molecular target of ovarian cancer treatment.Methods:(1)Ovarian cancer cells SKOV-3 were divided into experimental group and control group.It will be cultured in the SKOV-3 cell culture medium with the addition of HA for1440minutes;After having been cultured with the addition of anti-CD44 antibody for 1hour,it will be cultured for 1440 more minutes with the addition of HA.Western-blot was used to detect the expression of STAT3 protein in SKOV3 cells.(2)It will be cultured in the SKOV-3 cell culture medium with the addition of HA for 1440minutes;After having been cultured with the addition of anti-CD44 antibody for 60 minutes,it will be cultured for 1440 more minutes with the addition of HA;After having been cultured in the medium with the addition of Nanog small interfering RNA transfection reagent mixture for 2880 minutes,it will be cultured in for 1440 more minutes with the addition of HA;After having been cultured in the medium with the addition of Nanog complementation DNA transfection reagent mixture for 2880 minutes,it will be cultured in for 1440 more minutes with the addition of HA.RT-PCR method was used to detect the content of STAT-3messenger RNA in SKOV3 cells.(3)It will be cultured in the SKOV-3 cell culture medium with the addition of HA for 1440 more minutes;After having been cultured with the addition of anti-CD44 antibody for 60 minutes,it will be cultured for 1440 more minutes with the addition of HA;After having been cultured in the medium with the addition of Nanog small interferingRNA transfection reagent mixture for 2880 minutes,it will be cultured in for 1440 more minutes with the addition of HA.In addition,CCK-8was added to the cell culture medium for 60 minutes,and the absorbance value of 450 nm was measured by enzyme marker.Results:The relative expression level of STAT-3 protein in the control group;the addition of HA group;Add anti-CD44 antibody and then add HA group is 1.0945±0.0187 ?1.3233±0.0277?0.8519±0.0198.The difference was statistically significant(P<0.05).The relative expression level of STAT-3 messenger RNA in the control group;the addition of HA group;Add anti-CD44 antibody and then add HA group;Transfection of Nanog small interfering RNA and then add HA group;Transfection of Nanog complementation DNA and then add HA group is 2.0841±0.0391 ? 3.6100±0.3208 ? 1.2500±0.0800 ?1.8533±0.0833?6.9367±0.0961,The difference was statistically significant(P<0.05).The absorbance value of the control group;the addition of HA group;Add anti-CD44 antibody and then add HA group;Transfection of Nanog small interfering RNA and then add HA group;Transfection of Nanog complementation DNA and then add HA group is 0.584?0.847?0.380?0.464?1.145,The difference was statistically significant(P<0.05).Conclusion:In HA and CD44 signaling pathway,Nanog gene was over-expressed in ovarian cancer SKOV3 cell lines,and it has a synergistic effect with STAT3,which could promote the growth of ovarian cancer cells.