溴结构域蛋白4抑制剂JQ1促使肝癌细胞侵袭迁移的分子机制

The molecular mechanism of bromdomain protein 4 inhibitor JQ1 promoting the invasion and migration of hepatocellular carcinoma cells in vitro

目的探讨溴结构域蛋白4抑制剂JQ1对肝癌细胞增殖、侵袭和迁移能力的影响,并探究其相关作用机制。方法采用细胞增殖实验(MTS)检测JQ1对肝癌细胞增殖的影响,Transwell检测JQ1对肝癌细胞细胞迁移、侵袭能力的影响,蛋白免疫印迹法检测JQ1对肝癌细胞E-cadherin、N-cadherin、Vimentin、基质金属蛋白酶(MMP)-9/2表达情况;Transwell检测P-ERK1/2抑制剂(SCH772984)联合JQ1处理对肝癌细胞迁移、侵袭能力的变化,并用蛋白免疫印迹法检测JQ1联合P-ERK1/2抑制剂处理后MMP-9/2变化。结果 JQ1对肝癌细胞Hep3b、SMMC-7721增殖有抑制作用(P<0.05),JQ1能够促进肝癌细胞系Hep3b、SMMC-7721的MMP-9、P-ERK1/2表达,进而增强肝癌细胞系Hep3b、SMMC-7721迁移、侵袭能力(P均<0.001)。JQ1诱导的肝癌细胞的迁移、侵袭的能力的增强可以被P-ERK1/2抑制剂(SCH772984)抑制(P均<0.001)。结论 JQ1可以通过促进ERK1/2磷酸化进而促进MMP-9/2表达,进而促进肝癌细胞侵袭、迁移。

Objective To investigate the effect and mechanism of bromdomain protein4inhibitor JQ1on the proliferation,invasion and migration of hepatocellular carcinoma cells.Methods MTS assay wasperformed to evaluate the effect of JQ1on the proliferation of hepatocellular carcinoma cells.Transwell assay was conducted to assess the influence of JQ1on the invasion and migration of hepatocellular carcinoma cells.Western blot was carried out to measure the expression of E-cadherin,N-cadherin,Vimentin,matrix metalloproteinase(MMP)-9/2proteins.Transwell assay was adopted to assess the effect of combined treatment of P-ERK1/2inhibitor(SCH772984)and JQ1upon the invasion and migration of hepatocellular carcinoma cells.Western blot was used to detect the changes of MMP-9/2expression in hepatocellular carcinoma cells after treated with JQ1combined with P-ERK1/2inhibitor.Results JQ1could significantly inhibit the proliferation of hepatocellular carcinoma cells Hep3b and SMMC-7721(both P<0.05).JQ1could significantly up-regulate the expression of MMP-9and P-ERK1/2in Hep3b and SMMC-7721cells,thereby evidently enhancing the migration and invasion of Hep3b and SMMC-7721cells(all P<0.001).The effect of JQ1upon promoting the invasion and migration of hepatocellular carcinoma cells could be significantly inhibited by P-ERK1/2inhibitor of SCH772984(both P<0.001).Conclusions JQ1can promote the invasion and migration of hepatocellular carcinoma cells by promoting the phosphorylation of ERK1/2and up-regulating the expression of MMP-9/2.