对香豆酸通过Akt/TSC/mTOR通路对食管癌细胞增殖、凋亡及内质网应激分子信号表达的影响  

Effects of p-coumaric acid on the proliferation,apoptosis,and expression of endoplasmic reticulum stress molecule signal in esophageal cancer cells through Akt/TSC/mTOR

作　　者：周成如 姚志刚 杨维维 ZHOU Chengru;YAO Zhigang;YANG Weiwei(Haian Hospital of Traditional Chinese Medicine,Haian 226600,Jiangsu,China)

机构地区：[1]海安市中医院,江苏海安226600

出　　处：《现代中西医结合杂志》2025年第3期318-324,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：江苏省2022年度医学科学研究重点课题计划项目(202202626);南通市科学技术局项目(JCZ2022085)。

摘　　要：目的探究对香豆酸对食管癌细胞增殖、凋亡、内质网应激以及蛋白激酶B/结节性硬化症复合物蛋白/哺乳动物雷帕霉素靶蛋白(Akt/TSC/mTOR)信号通路的影响。方法使用不同浓度的对香豆酸(0~8 mmol/L)作用于人食管癌细胞株TE-1,采用CCK-8法检测细胞存活率,筛选最佳药物浓度。将TE-1细胞随机分为TE-1组、对香豆酸低浓度组(用含有0.5 mmol/L对香豆酸的培养基培养)、对香豆酸中浓度组(用含有1.0 mmol/L对香豆酸的培养基培养)、对香豆酸高浓度组(用含有2.0 mmol/L对香豆酸的培养基培养)和对香豆酸高浓度+IGF-1组(用含有2.0 mmol/L对香豆酸和10μmol/L IGF-1的培养基培养),采用克隆形成实验检测各组克隆形成细胞数量,采用流式细胞术检测各组细胞凋亡率,采用Transwell实验检测各组细胞迁移及侵袭能力,采用Western blot法检测各组细胞中Akt/TSC/mTOR通路蛋白(p-Akt/Akt、p-TSC/TSC、p-mTOR/mTOR)及内质网应激相关蛋白[蛋白激酶PKR样内质网激酶(PERK)、葡萄糖调节蛋白78(GRP78)、转录激活因子6(ATF6)]表达情况。结果与TE-1组比较,对香豆酸低、中、高浓度组细胞克隆形成数量、迁移及侵袭细胞数量均逐渐减少(P均<0.05),细胞中p-Akt/Akt、p-TSC/TSC、p-mTOR/mTOR相对表达量均逐渐降低(P均<0.05),细胞凋亡率及细胞中PERK、GRP78、ATF6蛋白相对表达量均逐渐升高(P均<0.05);与对香豆酸高浓度组比较,对香豆酸高浓度+IGF-1组细胞克隆形成数量、迁移及侵袭细胞数量均明显增加(P均<0.05),细胞中p-Akt/Akt、p-TSC/TSC、p-mTOR/mTOR相对表达量均明显升高(P均<0.05),细胞凋亡率及细胞中PERK、GRP78、ATF6蛋白相对表达量均明显降低(P均<0.05)。结论对香豆酸能够抑制食管癌细胞增殖、迁移与侵袭,促进癌细胞内质网应激及凋亡的发生,其作用机制可能与抑制Akt/TSC/mTOR信号通路有关。Objective It is to investigate the effects of p-coumaric acid(p-CA)on proliferation,apoptosis,endoplasmic reticulum stress,and protein kinase B/tuberous sclerosis complex/mammalian target of rapamycin(Akt/TSC/mTOR)signaling pathway in esophageal cancer cells.Methods The human esophageal cancer cell line TE-1 were treated with different concentrations of p-CA(0~8 mmol/L),and the cell viability was detected by CCK-8 assay to the optimal drug concentration.The TE-1 cells were randomly divided into TE-1 group,p-CA low concentration group(cultured with medium containing 0.5 mmol/L p-CA),p-CA medium concentration group(cultured with medium containing 1.0 mmol/L p-CA),p-CA high concentration group(cultured with medium containing 2.0 mmol/L p-CA),and p-CA high concentration+IGF-1 group(cultured with medium containing 2.0 mmol/L p-CA and 10μmol/L IGF-1).The number of clone-forming cells in each group was detected by clone formation assay,the apoptosis rate of cells in each group was detected by flow cytometry,the migration and invasion abilities of cells in each group were detected by Transwell assay,and the expressions of Akt/tSC/mTOR pathway proteins(p-Akt/Akt,p-TSC/TSC,p-mTOR/mTOR)and endoplasmic reticulum stress-related proteins[PKR-like endoplasmic reticulum kinase(PERK),glucose-regulated protein 78(GRP78),and activator of transcription factor 6(ATF6)]in the cells of each group were detected by Western blotting.Results Compared with the TE-1 group,the numbers of cell clone formation,migration and invasion cells in the p-CA low,medium and high concentration groups were gradually reduced(all P<0.05),the relative expressions of p-Akt/Akt,p-TSC/TSC,p-mTOR/mTOR in the cells were gradually decreased(all P<0.05),the apoptosis rate and the cellular protein relative expressions of PERK,GRP78,ATF6 were gradually increased(all P<0.05);compared with the p-CA high concentration group,the numbers of cell clone formation,migration and invasion cells in the p-CA+IGF-1 group were significantly increased(all P<0.05),the relative expr

关 键 词：食管癌 对香豆酸 蛋白激酶B/结节性硬化症复合物蛋白/哺乳动物雷帕霉素靶蛋白信号通路 增殖 凋亡 内质网应激 

分 类 号：R735.1[医药卫生—肿瘤]