Egr-1对低氧应激下肝癌细胞BEL-7402黏附和迁移能力的影响
Egr-1 Regulates Human Hepatocelluar Carcinoma Cell Migration and Adhesion
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摘要: 目的 探讨显性负性突变（dominant negative）抑制即刻早期蛋白Egr-1转录因子活性对低氧应激下肝癌BEL-7402细胞黏附和迁移能力的影响。方法构建显性负性突变Egr-1腺病毒，感染BEL-7402细胞，同时以空载体和空白细胞做对照。“划痕”实验检测细胞迁移能力的变化；细胞与基质黏附实验，免疫荧光检测Egr-1对黏附能力的影响；Westernblot检测相关蛋白的改变。结果成功构建获得Egr-1转录功能竞争抑制型稳定腺病毒，并感染BEL-7402细胞。与BEL-7402细胞比较，低氧状态下Ad-dnEgr-1/BEL-7402组细胞，迁移，黏附能力显著降低（P<0.01），细胞中与迁移、粘附相关的微丝所形成的伪足结构明显减少，FAK蛋白磷酸化水平降低。结论抑制Egr-1转录因子功能可以抑制肝癌BEL-7402细胞的迁移和黏附。Abstract: Objective To investigate the effect of dominant negative mutation of Egr-1 expression on the adhension and invasion in the human Hepatocellular carcinoma cell line BEL-7402. Methods Adenovirus of Egr-1 with dominant negative mutation was constructed and transfected the BEL-7402 cell line.Empty adenovirus-transfected and normal BEL-7402 were used as control groups.The influences of Egr-1 on cell migration and adhesion capacity were detected by cell scratch test and adhesion test. Results Transcription function competitive inhibition-type adenovirus of Egr-1 was successfully constructed and transfected into BEL-7402 cells. Compared with control BEL-7402 cells，both migration and adhesion of Ad-dnEgr-1/BEL-7402 cells were significantly lower (P<0.01).Pseudopod structure related to cell migration and adhesion significantly reduced，and FAK phosphorylation levels decreased under hypoxic conditions. Conclusion Down-regulation of Egr-1 transcription function could inhibit the proliferation and invasive of BEL-7402 cells.
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