Sphk1对人结肠癌细胞株Lovo增殖与侵袭的影响及其作用机制  

Role of Sphk1 in the proliferation and invasion of human colon cancer Lovo cells

作　　者：钟月圆[1] 黄杰安[1] 刘诗权[1] 覃蒙斌[1] 金卉[1] 

机构地区：[1]广西医科大学第一附属医院消化内科,广西壮族自治区南宁市530021

出　　处：《世界华人消化杂志》2010年第24期2528-2532,共5页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30760275;广西科学基金资助项目;No.0832008~~

摘　　要：目的:研究Sphk1对结肠癌细胞增殖、凋亡及侵袭的影响并探讨其机制.方法:将人结肠癌Lovo细胞株分成Sphk1激活组,Sphk1抑制组,空白对照组.以佛波醇-12-豆蔻酸酯-13-乙酸酯(phorbol12-myristate13-acetate,PMA)为Sphk1激活剂(终浓度为100nmol/L),N,N-二甲基鞘胺醇(erythro-sphingo-sineN,N-Dimethyl,DMS)为Sphk1抑制剂(终浓度为50μmol/L)处理Lovo细胞24h后,用MTT方法测定细胞的增殖活性,用流式细胞术检测细胞凋亡,用Transwell侵袭实验检测细胞侵袭能力,用Westernblot测定细胞Sphk1、ERK1/2、p-ERK1/2、NF-κBp65蛋白水平的变化.结果:PMA可以明显诱导Sphk1蛋白的表达,促进Lovo细胞生长,抑制细胞的凋亡,并促进细胞的侵袭;相反,DMS明显抑制Sphk1的表达,抑制细胞生长,促进细胞的凋亡,并抑制细胞的侵袭.Sphk1激活组、对照组、抑制组的细胞凋亡率分别是9.15%,16.25%,32.58%.与对照组相比,Sphk1激活组、抑制组的细胞相对侵袭率分别是190.57%,9.65%,差异有统计学意义(均P<0.01).PMA诱导Sphk1表达,同时伴有ERK1/2、p-ERK1/2和NF-κBp65蛋白表达的上调,DMS抑制Sphk1表达,可抑制ERK1/2、p-ERK1/2、NF-κBp65蛋白的表达.结论:Sphk1可促进Lovo细胞的生长增殖与侵袭并抑制细胞的凋亡,其机制可能与ERK1/2和NF-κB信号通路的激活有关.AIM：To investigate the role of sphingosine kinase 1（Sphk1）in the proliferation,apoptosis and invasion of colon cancer cells.METHODS：Human colon cancer Lovo cells were divided into three groups：Sphk1 activation group [treated with 100 nmol/L phorbol 12-myristate 13-acetate（PMA）],Sphk1 suppression group [treated with 50 μmol/L N,Ndimethyl-D-erythro-sphingosine（DMS）],and control group（treated with 9 g/L NaCl）.Cell proliferation activity was detected by MTT assay;cell apoptosis was detected by flow cytometry;cell invasion was detected by Transwell chamber assay;and the protein expression of Sphk1,ERK1/2,p-ERK1/2,and NF-κB p65 was detected by Western blot.RESULTS：PMA significantly induced the expression of Sphk1 protein,promoted Lovo cell growth and invasion,inhibited cell apoptosis,and up-regulated the protein expression of ERK1/2,p-ERK1/2,and NF-κB p65.In contrast,DMS significantly inhibited the expression of Sphk1 protein,suppressed cell growth,promoted apoptosis,and down-regulated the protein expression of ERK1/2,p-ERK1/2,and NF-κB p65.The apoptosis rates in the Sphk1 activation group,Sphk1 suppression group and control group were 9.15%,16.25% and 32.58%,respectively.The relative invasion rate in the Sphk1 activation group was significantly higher than that in the Sphk1 suppression group（190.57% vs 9.65%,P 0.01）.CONCLUSION：Sphk1 promotes the proliferation and invasion but inhibits apoptosis of Lovo cells possibly via a mechanism associated with the activation of ERK1/2 and NF-κB signaling pathways.

关 键 词：鞘氨醇激酶-1 人结肠癌细胞 侵袭 凋亡 增殖 

分 类 号：R735.35[医药卫生—肿瘤]