摘要
目的 探究锌-α2-糖蛋白1(AZGP1)对食管鳞癌细胞增殖、凋亡及化疗敏感性的调控作用,并探讨其可能的作用机制.方法 采用实时荧光定量聚合酶链反应(qRT-PCR)和Western blotting检测AZGP1在人正常食管上皮细胞(Het-1A)和人食管鳞癌细胞(TE11、EC9706、HCE7、EC109、KYSE150)中的表达.构建AZGP1过表达质粒并转染至EC9706细胞,分为对照组、过表达对照组(Ov-NC组)、AZGP1过表达组(Ov-AZGP1组);将AZGP1过表达质粒与脂肪酸合酶(FASN)过表达质粒共转染至EC9706细胞或将AZGP1过表达质粒单独转染至经Wnt通路激活剂Wnt agonist 1处理的EC9706细胞,分为对照组、Ov-AZGP1组、Ov-AZGP1+Ov-FASN组、Ov-AZGP1+Wnt agonist 1组..CCK-8法和克隆形成实验检测细胞增殖;TUNEL实验检测细胞凋亡;CCK-8法和TUNEL实验检测顺铂和5-氟尿嘧啶化疗敏感性;Western blotting检测凋亡及FASN/Wnt/β-catenin通路相关蛋白的表达.结果 AZGP1在食管鳞癌细胞中表达降低(P＜0.05),AZGP1过表达可抑制细胞增殖(P＜0.05)、促进细胞凋亡(P＜0.05)并增加顺铂和5-氟尿嘧啶化疗敏感性(P＜0.05).AZGP1过表达可抑制FASN/Wnt/β-catenin信号通路(P＜0.05),FASN过表达或Wnt agonist 1可逆转AZGP1对EC9706细胞增殖(P＜0.05)、凋亡(P＜0.05)及化疗敏感性的作用(P＜0.05).结论 AZGP1可通过抑制FASN/Wnt/β-catenin通路进而抑制食管鳞癌细胞增殖、诱导细胞凋亡并增加化疗敏感性.
Abstract
Objective To investigate the regulatory role of Zinc-α2-glycoprotein 1(AZGP1)in proliferation,apoptosis,and chemotherapy sensitivity of esophageal squamous cell carcinoma(ESCC)cells,and explore its possible mechanism of action.Methods The expression of AZGP1 in human normal esophageal epithelial cells(Het-1A)and ESCC cells(TE11,EC9706,HCE7,EC109,KYSE150)was detected by real-time quantitative polymerase chain reaction(qRT-PCR)and Western blotting.An AZGP1 overexpression plasmid was constructed and transfected into EC9706 cells,which were divided into control group,overexpression negative control group(Ov-NC),and AZGP1 overexpression group(Ov-AZGP1 group).The AZGP1 overexpression plasmid was transfected alone or in combination with a fatty acid synthase(FASN)overexpression plasmid into EC9706 cells treated with Wnt pathway activator Wnt agonist 1,divided into control group,Ov-AZGP1 group,Ov-FASN group,Ov-AZGP1+ Ov-FASN group,and Ov-AZGP1+Wnt agonist 1 group.Cell proliferation was detected by CCK-8 assay and colony formation assay.Cell apoptosis was detected by TUNEL assay.Chemotherapy sensitivity to cisplatin and 5-fluorouracil was evaluated by CCK-8 assay and TUNEL assay.The expression of apoptosis-related proteins and FASN/Wnt/β-catenin pathway-related proteins was detected by Western blotting.Results AZGP1 expression was downregulated in ESCC cells(P＜0.05).Overexpression of AZGP1 inhibited cell proliferation(P＜0.05),promoted apoptosis(P＜0.05),and increased sensitivity to cisplatin and 5-fluorouracil chemotherapy(P＜0.05).Overexpression of AZGP1 inhibited the FASN/Wnt/β-catenin signaling pathway(P＜0.05),and overexpression of FASN or Wnt agonist 1 reversed the effects of AZGP1 on cell proliferation(P＜0.05),apoptosis(P＜0.05),and chemotherapy sensitivity(P＜0.05)in EC9706 cells.Conclusion AZGP1 can inhibit the proliferation,induce apoptosis,and increase chemotherapy sensitivity of ESCC cells by suppressing the FASN/Wnt/β-catenin pathway.
维普
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