全反式视黄酸衍生物逆转高水平胰岛素对胃癌细胞BGC-823迁移的作用研究  

All-trans retinoic acid derivative reverses the effect of high-level insulin on the migration of human gastric cancer cell BGC-823

作　　者：王陈 胡安拉[1] 赵奇红[1] 汪渊 张素梅 Wang Chen;Hu Anla;Zhao Qihong(Dept of Nutrition and Food Hygiene,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学公共卫生学院营养与食品卫生学系,合肥230032 [2]安徽医科大学基础医学院生物化学教研室,合肥230032

出　　处：《安徽医科大学学报》2021年第9期1390-1396,共7页Acta Universitatis Medicinalis Anhui

基　　金：安徽高校自然科学研究项目(编号:KJ2019A0227);国家自然科学基金(编号:81272399)。

摘　　要：目的探讨高水平胰岛素条件下全反式视黄酸衍生物4-氨基-2-三氟甲基-苯基-维甲酸酯(ATPR)对人胃癌细胞BGC-823迁移能力的影响及其潜在的分子机制。方法首先采用不同浓度胰岛素刺激BGC-823细胞,CCK-8法检测胰岛素对细胞增殖能力的影响;然后采用等同人高胰岛素血症水平(≥0.5 ng/ml)而对BGC-823增殖能力没有影响的胰岛素剂量(<10.0 ng/ml),联合ATPR处理BGC-823细胞后,通过细胞划痕实验和Transwell实验明确BGC-823细胞迁移能力的改变;Western blot检测与细胞迁移能力相关蛋白的表达水平。结果CCK-8结果显示,与对照组比较,0.5、5.0 ng/ml胰岛素作用48 h后对BGC-823细胞的增殖无影响,而10.0 ng/ml胰岛素能够促进BGC-823细胞增殖;细胞划痕和Transwell实验显示,5.0 ng/ml胰岛素明显促进BGC-823细胞的迁移能力;但是,联合ATPR后能够抑制胰岛素对BGC-823迁移的促进作用,作用与肌球蛋白轻链激酶(MLCK)抑制剂ML-7类似;Western blot结果显示胰岛素促进MLCK蛋白表达且下调胃特异性紧密连接蛋白Claudin 18的表达,而ATPR联合处理后能抑制MLCK并促进Claudin 18蛋白的表达。结论ATPR可逆转高水平胰岛素对胃癌细胞BGC-823迁移的促进作用,可能与其抑制胰岛素对MLCK和Claudin 18的调控密切相关。Objective To explore the impact of 4-amino-2-trifluoromethyl-phenyl retinate(ATPR),a derivative of all-trans retinoic acid,on the motility of human gastric cancer cell line BGC-823 and the possible mechanismsinvolved under high-level insulin.Methods Firstly,BGC-823 cells were treated with various concentrations of insulin.Cell counting kit-8(CCK-8)assay was used to detect the effect of insulin on cell proliferation.Then,BGC-823 cells were treated with ATPR combined with different concentrations(with no effect on cell growth but could simulate the high insulin level of humans)of insulin.Wound healing and Transwell assays were used to measure the motility of BGC-823 cells under different treatments.Finally,Western blot assay was performed to detect the expression of proteins related to cell migration.Results CCK-8 results showed that,compared with the control group,0.5 ng/ml and 5.0 ng/ml insulin had no effect on proliferation of BGC-823 cells after 48 hours while 10.0 ng/ml insulin significantly promoted the cell proliferation.Wound healing and Transwell assay showed that 5.0 ng/ml of insulin promoted the migration of BGC-823 cells,however,ATPR reversed the effect of insulin on BGC-823 cells motility,which could also be attenuated by a specific inhibitor of myosin light chain kinase(MLCK)-ML-7.Western blot revealed that insulin upregulated the expression of MLCK while downregulated Claudin 18.But,after ATPR treated,the protein expression of MLCK was inhibited while the expression of Claudin 18 increased.Conclusion ATPR can reverse the effect of high-level insulin on the motility of BGC-823 cells,which may be attributed to the regulation of ATPR on MLCK and Claudin 18 under high-level insulin.

关 键 词：ATPR 胃癌 迁移 胰岛素 

分 类 号：R735.2[医药卫生—肿瘤]