内质网核糖体结合蛋白1表达上调对肝癌细胞迁移与侵袭的影响

Effect of Overexpression of RRBP1 on Metastasis and Invasion of Liver Cancer Cells

目的探讨内质网核糖体结合蛋白1(RRBP1)在促进肝癌转移中的调控作用。方法1)分别用生物信息学分析方法与免疫组化实验,分析RRBP1在原发性与转移性肝癌组织中的表达。2)通过转染siRNA下调RRBP1表达后,分别用划痕实验与Transwell迁移实验分析对肝癌细胞运动与迁移能力的影响。3)通过转染siRNA下调RRBP1表达后,用Transwell侵袭实验分析对肝癌细胞侵袭能力的影响。4)通过转染siRNA下调RRBP1表达后,用RT-PCR与蛋白质印迹技术实验检测对肝癌细胞上皮间质转化(EMT)标志分子表达的影响,进而分析RRBP1在肝癌细胞EMT中的调控作用。结果1)生物信息分析表明,RRBP1在肝癌组织尤其是转移性肝癌组织中,表达高于正常肝组织;免疫组化实验也发现,RRBP1在肝癌转移灶组织中的表达高于原发灶。2)下调RRBP1减弱了肝癌细胞的运动与迁移能力。3)下调RRBP1抑制了肝癌细胞的侵袭。4)下调RRBP1表达升高了上皮细胞标志分子E-cadherin与ZO-1的表达,却降低了间质细胞标志分子N-cadherin与Vimentin的表达。结论RRBP1在肝癌转移中发挥了重要的促进作用,EMT增强可能是RRBP1促进肝癌转移的主要机制。

Objective To investigate the role of endoplasmic reticulum ribosome binding protein 1(RRBP1)in promoting the metastasis of liver cancer.Methods(1)The expression of RRBP1 in primary and metastatic liver cancer tissues was analyzed by bioinformatics analysis and immunohistochemical experiments.(2)The effects of RRBP1 knockdown by siRNA transfection on the metastasis of liver cancer cells were determined by wound healing and transwell migration assays.(3)The effects of RRBP1 knockdown by siRNA transfection on the invasion of liver cancer cells were determined by transwell invasion assay.(4)The effects of RRBP1 knockdown by siRNA transfection on the expressions of markers involved in epithelial-mesenchymal transition(EMT)of liver cancer cells were determined by RT-PCR and Western blotting analysis.Results(1)Bioinformatic analysis indicated that RRBP1 expression increased in tumor tissues,especially in metastasis tumor tissues,of liver cancer compared with normal liver tissues.Immunohistochemistry assay also showed that RRBP1 expression was higher in the metastatic liver cancer tissues than in primary liver cancer tissues.(2)RRBP1 knockdown significantly suppressed the metastasis ability of liver cancer cells.(3)RRBP1 knockdown also markedly suppressed the invasion ability of liver cancer cells.(4)RRBP1 knockdown increased the expression of epithelial markers of E-cadherin and ZO-1,while decreased the expression of mesenchymal markers of N-cadherin and Vimentin.Conclusion RRBP1 plays a crucial role in liver cancer metastasis.RRBP1 promotes the metastasis of liver cancer mainly through induction of EMT.