AZGP1通过FASN/Wnt/β-catenin通路对食管鳞癌细胞增殖、凋亡及化疗敏感性的影响

Effect of AZGP1 on proliferation, apoptosis and chemosensitivity of esophageal squamous cell carcinoma cells via FASN/Wnt/β-catenin signaling

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.