ACSS2 Mediates The Cisplatin Sensitivity Of Human Esophageal Squamous Cell Carcinoma Via PI3K/AKT Pathway

?Objective?To explore the effect of the expression change of acetyl-CoA synthetase 2(ACSS2)on the sensitivity of esophageal squamous cell carcinoma(ESCC)to cisplatin(DDP)and relevant mechanism.?Methods?1)ACSS2 expression was detected in tumor and paracancerous tissues collected from 40 cases of ESCC by immunohistochemistry and western blot.In vitro verification of ACSS2 expression difference was performed by western blot in ESCC cell line and normal esophageal squamous Het-1A cell line.2)ACSS2 downregulated and overexpressed cell strains were constructed by liposome transfection of siRNA-ACSS2 or lentivirus transfection of LV-ACSS2 into TE-1 cells.CCK8 assay was used to detect the change of IC50 value of DDP by downregulating the expression of ACSS2.3)The effect of ACSS2 interference on TE-1 cell apoptosis was tested by flow cytometry before and after DDP treatment(5 ?g/mL).4)western blot was carried out to detect the expression of proteins related to PI3K/AKT signaling pathway and apoptotic-related protein cleaved-Caspase-3.?Results?1)Immunohistochemistry and western blot results showed that the expression intensity of ACSS2 in ESCC tissue was significantly higher than that in paracancerous normal tissue(P<0.001).In vitro test indicated that the expression of ACSS2 in ESCC cell line was obviously higher than that of normal esophageal squamous Het-1A cell line.2)CCK8 results showed that IC50 of TE-1 cells was remarkably downregulated after siRNA-ACSS2 treatment(all P<0.001).3)Flow cytometry results showed that the apoptosis rate of ESCC cells was obviously decreased by inhibiting ACSS2 expression(P<0.05),which,however,increased significantly under the combined treatment of 5 ?g/mL DDP and ACSS2interference(all P<0.001).4)Western blotting results revealed that cisplatin treatment up-regulated the expression of ACSS2,p-PI3 K and p-AKT in TE-1 cells.Under the action of cisplatin,after targeted inhibition of ACSS2,p-PI3 K and p-AKT were significantly reduced and the expression of apoptosis-related protein cleaved-Caspase-3 was significantly increased,overexpression of ACSS2 can effectively reverse the formation of cleaved-Caspase-3 while maintaining PI3 K / AKT signal activation.?Conclusions?ACSS2 protein regulates cisplatin sensitivity of esophageal squamous cell carcinoma by participating in PI3 K / AKT signaling pathway activation and cleaved-Caspase-3 expression.