鞘鞍醇激酶1对肝癌细胞凋亡特性的影响

摘要

Objective: To investigate the role of sphingosine kinase 1 (SPK1) on the apoptosis of human hepatocellular carcinoma cell line BEL-FU. Methods:The adenovirus carrying the wild type SPK1 gene(SPK1WT) and SiRNASPK1 (Ad-H1-SPK1) were constructed respectively;viral infection titer was determined by TCID50 and CPE. Methods: Cellular SPK enzyme activity was assayed; the effect of SPK1 on the apoptosis of BEL-FU was evaluated by MTT. Results: Re-adenoviruses were amplified and purified; SPK enzyme activity assay suggested SPKWT could increase SPK activity and suppress the effect of DMS on BEL-FU, while Ad-H1-SPK1 could decrease SPK activity respectively and enhance the effect of DMS on BEL-FU. Conclusion: SiRNASPK1 could become a new method for hepatocellular carcinoma treatment.%目的:研究鞘鞍醇激酶1(SPK1)对人肝癌耐药细胞株BEL-FU凋亡特性的影响.方法:扩增并纯化携带野生型(Ad-SPK1WT)和SiRNASPK1(Ad-H1-SPK1)的重组腺病毒,以TCID50方法(1-U/ml)和快速CPE法测定病毒感染滴度;以腺病毒为载体将SPK1基因导入肝癌细胞BEL-FU;分别以酶活性检测试剂盒、MTT法检测SPK酶活性及SPK1对肝癌细胞凋亡特性的影晌.结果:获得高滴度的Ad-SPK1WT腺病毒和Ad-H1-SPK1腺病毒;携带Ad-SPK1WT基因的BEL-FU细胞中SPK1酶活性增强并抑制DMS(N,N,-二甲基鞘氨醇)对BEL-FU的细胞毒作用,抑制肿瘤细胞凋亡;携带Ad-H1-SPK1基因的BEL-FU细胞中SPK1酶活性低并增强DMS(N,N,-二甲基鞘氨醇)对BEL-FU的细胞毒作用,促进肿瘤细胞凋亡.结论:阻断SPK1作用可能成为肝癌治疗的新途径.