This aim of this article was to investigate the effects of SFTN(sufentanil)modulating cAMP(cyclic adenosine monophosphate)/PKA(protein kinase B)/CREB(cAMP-response element binding protein)sig-naling pathway on the proliferation,apoptosis and invasion of OC(ovarian cancer)cells.Human SKOV-3(OC cells)were treated with sufentanil at concentrations ranging from 2.5 to 160 ng/mL,and cell activity was detected by the CCK-8 assay to screen for optimal drug concentration.SKOV-3 cells were divided into Control group,sufentanil low,medium,and high concentration groups(SFTN-L group,SFTN-M group,SFTN-H group),and sufentanil high concentration+PKA activator group(SFTN-H+8-bromo-cAMP group);and the plate cloning assay was used to detect cell proliferation;the apoptosis was detected by flow cytometry;the migration was detected by the scratch assay;the invasion was detected by the Transwell assay.ELISA method was applied to detect cAMP level.Western blot method was applied to detect Ki67(nuclear proliferative antigen markers),cyclin D1,Caspase-3,Bax(B-cell lymphoma as-sociated X-protein),MMP-2(matrix metalloproteinase-2),MMP-9(matrix metalloproteinase-9),PKA(protein kinase B),p-PKA(phosphorylated protein kinase B),CREB,p-CREB(phosphorylated-CREB)protein expression.Nude mice transplantation tumor assay to detect the effect of sufentanil on the growth of OC transplantation tumor.The ef-fect of sufentanil on the growth of OC transplanted tumors was detected in nude mice transplantation tumor assay.The concentrations of sufentanil were selected as 20 ng/mL,40 ng/mL,and 80 ng/mL for subsequent experiments.Com-pared with the Control group,the number of colony formation,cell scratch healing rate,number of cell invasion and the expression levels of Ki67,Cyclin D1,MMP-2,MMP-9,cAMP,p-PKA/PKA,andp-CREB/CREB were decreased in the SFTN-L,SFTN-M,and SFTN-H groups,and the apoptosis rate and the Caspase-3,Bax expression levels were significantly increased in a concentration-dependent manner(P＜0.05);compared with the SFTN-H group,the num-ber of colony formation,the scratch healing rate,the number of cell invasion and the expression of Ki67,CyclinD1,MMP-2,MMP-9,cAMP,p-PKA/PKA,p-CREB/CREB in the SFTN-H+8-bromo-cAMP group were elevated,and the apoptosis rate and Caspase-3 and Bax expression levels were significantly reduced(P＜0.05).The transplantation tu-mor experiment showed that the transplantation tumors of mice in the SFTN group grew more slowly than those in the Control group,the mass and volume of the transplantation tumors were reduced,and the expression levels of cAMP,p-PKA/PKA,and p-CREB/CREB were decreased(P＜0.05).Inhibition of cAMP/PKA/CREB signaling pathway by sufentanil inhibited OC cell proliferation and invasion and promoted apoptosis.
ovarian cancersufentanilcAMP(cyclic adenosine monophosphate)/PKA(protein kinase B)/CREB(cAMP-response element binding protein)signaling pathwayproliferationapoptosisinvasion
NETL
NSTL
万方数据
