CAJ | 학술논문

目的研究鞘氨醇激酶1(SPK1)对人肝癌耐药细胞株BEL-FU生物学特性的影响.方法将SPK1siRNA基因(Ad-H 1-SPK1)及野生型SPK1基因(Ad-SPK1wT)导入人肝癌耐药细胞株BEL-FU.用试剂盒检测SPK酶活性,MTT法检测其对肝癌细胞凋亡的影响,Transwell法测定肝癌细胞侵袭力,Western-blot方法检测多药耐药相关蛋白1(MRP1)表达水平的变化.结果导入Ad-SPK1wT增强肝癌细胞SPK活性,转染SPK1 siRNA(Ad-H 1-SPK1)抑制肝癌细胞SPK活性；肝癌细胞导入SPK1后,抑制SPK特异性抑制剂N,N,-二甲基鞘氨醇(Dimethyl sphingosine,DMS)引起的肝癌细胞凋亡并促进肝癌细胞迁移和增加多药耐药蛋白( MRP1)的表达；阻断SPK1则显著促进细胞凋亡和明显抑制肝癌细胞迁移,且多药耐药蛋白(MRP1)表达量明显降低.结论 SPK1的活性与肝癌细胞的凋亡、侵袭力及多药耐药密切相关,阻断SPK1可为肝癌治疗提供新的靶点和策略.
목적 연구초안순격매1(SPK1)대인간암내약세포주BEL-FU생물학특성적영향.방법 장SPK1siRNA기인(Ad-H 1-SPK1)급야생형SPK1기인(Ad-SPK1wT)도입인간암내약세포주BEL-FU.용시제합검측SPK매활성,MTT법검측기대간암세포조망적영향,Transwell법측정간암세포침습력,Western-blot방법검측다약내약상관단백1(MRP1)표체수평적변화.결과 도입Ad-SPK1wT증강간암세포SPK활성,전염SPK1 siRNA(Ad-H 1-SPK1)억제간암세포SPK활성；간암세포도입SPK1후,억제SPK특이성억제제N,N,-이갑기초안순(Dimethyl sphingosine,DMS)인기적간암세포조망병촉진간암세포천이화증가다약내약단백( MRP1)적표체；조단SPK1칙현저촉진세포조망화명현억제간암세포천이,차다약내약단백(MRP1)표체량명현강저.결론 SPK1적활성여간암세포적조망、침습력급다약내약밀절상관,조단SPK1가위간암치료제공신적파점화책략.
Objective To investigate the roles of sphingosine kinasel (SPK1) in apoptosis,invasiveness and multidrug resistance of human hepatocellular carcinoma cell line BEL-FU.Methods BEL-FU cells were infected with adenovirus carrying SPK1wT gene and SPK1siRNA (Ad-H1-SPK1) gene.Their effects on biological characteristics of BEL-FU cells were evaluated by MTT,cellular SPK enzyme activity assay,Transwell Migration Technology and Western-blot,respectively.Results AdSPK1wT significantly increased SPK activity but SPK1siRNA(Ad-H1-SPK1) decreased SPK activity.Over expression of SPK1 suppressed the apoptosis induced by DMS(Dimethyl sphingosine,DMS) and enhanced migration of BEL-FU cells.The cells infected with SPK1 siRNA( Ad-H1-SPK1)significantly increased the apoptosis induced by DMS and inhibited the migration of human hepatocellular carcinoma cells.The expression of multidrug resistance-related protein (MRP1) of cells infected with SPK1siRNA (Ad-H1-SPK1) was suppressed significantly compared with the control group,while the expression of MRP1 infected with Ad- SPK1wT was enhanced.Conclusion SPK1 activity is closely associated with apoptosis、migration and multidrug resistance of human hepatocellular carcinoma cells,therefore,it may serve as a new target for HCC treatment.