扁蒴藤素提高人食管鳞状细胞癌耐药细胞株ECA109/DDP的顺铂敏感性OA

Objective To investigate the mechanism by which pristimerin enhances cis-diamine dichloroplatinum/cisplatin(DDP)sensitivity of esophageal squamous cell carcinoma cell line ECA109.Methods ECA109 cells were treated with 20 μg/mL DDP,with drug concentrations incrementally increased(40,80,100 μg/mL)every 2-3 passages.After 3-6 months,stable DDP-resistant ECA109/DDP cells were obtained which proliferated in high DDP concentration environment(80 μg/mL).The ECA109/DDP cells were divided into the following groups:control,DDP(80 μg/mL),pristimerin(1 μmol/L),pristimerin(1 μmol/L)+DDP(80 μg/mL),and pristimerin(1 μmol/L)+DDP(80 μg/mL)+TGF-β activator(SRI-011381)(10 μmol/L).Cell proliferation was assessed by CCK-8 assay and colony formation test;Apoptosis was examined by flow cytometry;Migration and invasion were evaluated via scratch wound healing and Transwell assays,respectively.The protein expressions of multidrug re-sistance-associated protein 1(MRP1),P-glycoprotein(P-gp),TGF-β1,Smad4,and CD44 were measured by Western blot.Results Compared with control group,there was no difference in various indicators in the DDP group(P＜0.05).While pristimerin group exhibited a decrease of A450 values,colony formation rate,scratch healing rate,number of invasive cells.The protein level of MRP1,P-gp,TGF-β1,Smad4,and CD44 also de-creased,along with increased apoptosis(P＜0.05).Compared with the DDP and pristimerin groups,the pristim-erin+DDP group showed further reduction in A450 values,colony formation rate,scratch healing rate,number of invasive cells,and protein level of MRP1,P-gp,TGF-β1,Smad4,and CD44,as well as increased apoptosis(P＜0.05).Compared with pristimerin+DDP group,pristimerin+DDP+SRI-011381 group showed an opposite trend in all the above indicators(P＜0.05).Conclusions Pristimerin enhances DDP sensitivity in ECA109/DDP cells by inhibition of TGF-β/Smad pathway.