Effects of Sufentanil on Proliferation, Apoptosis, and Invasion of OvarianCancer Cells by Regulating the cAMP/PKA/CREB Signaling Pathway

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) signaling 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 theCCK-8 assay to screen for optimal drug concentration. SKOV-3 cells were divided into Control group, sufentanillow, medium, and high concentration groups (SFTN-L group, SFTN-M group, SFTN-H group), and sufentanil highconcentration+PKA activator group (SFTN-H+8-bromo-cAMP group); and the plate cloning assay was used to detectcell proliferation; the apoptosis was detected by flow cytometry; the migration was detected by the scratch assay; theinvasion was detected by the Transwell assay. ELISA method was applied to detect cAMP level. Western blot methodwas applied to detect Ki67 (nuclear proliferative antigen markers), cyclin D1, Caspase-3, Bax (B-cell lymphoma associated X-protein), MMP-2 (matrix metalloproteinase-2), MMP-9 (matrix metalloproteinase-9), PKA (protein kinaseB), p-PKA (phosphorylated protein kinase B), CREB, p-CREB (phosphorylated-CREB) protein expression. Nudemice transplantation tumor assay to detect the effect of sufentanil on the growth of OC transplantation tumor. The effect of sufentanil on the growth of OC transplanted tumors was detected in nude mice transplantation tumor assay. Theconcentrations of sufentanil were selected as 20 ng/mL, 40 ng/mL, and 80 ng/mL for subsequent experiments. Compared with the Control group, the number of colony formation, cell scratch healing rate, number of cell invasion andthe expression levels of Ki67, Cyclin D1, MMP-2, MMP-9, cAMP, p-PKA/PKA, and p-CREB/CREB were decreasedin the SFTN-L, SFTN-M, and SFTN-H groups, and the apoptosis rate and the Caspase-3, Bax expression levels weresignificantly increased in a concentration-dependent manner (P<0.05); compared with the SFTN-H group, the number 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 theapoptosis rate and Caspase-3 and Bax expression levels were significantly reduced (P<0.05). The transplantation tumor experiment showed that the transplantation tumors of mice in the SFTN group grew more slowly than those in theControl 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 bysufentanil inhibited OC cell proliferation and invasion and promoted apoptosis.