鞘氨醇激酶1对结肠癌细胞血管生成拟态的影响及其机制  

Sphingosine kinase 1 induces vasculogenic mimicry formation by up-regulating VEGF expression and secretion in human colon cancer cell line HT-29

作　　者：李梦婷[1] 黄杰安[1] 周巧[1] 苏颖洁[1] 刘诗权[1] 覃蒙斌[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》2012年第33期3211-3217,共7页World Chinese Journal of Digestology

基　　金：广西卫生厅基金资助项目;桂卫重No.2010016;广西卫生厅基金资助项目;桂卫No.GZKZ10-107~~

摘　　要：目的:研究鞘氨醇激酶1(sphingosine kinase1,Sphk1)对人结肠癌HT-29细胞生成血管拟态(vasculogenic mimicry,VM)的影响及其可能的机制.方法:将人结肠癌HT-29细胞分为Sphk1抑制组、Sphk1激活组、对照组.Sphk1抑制组加入N,N-二甲基鞘氨醇(N,N-dimethyl-D-erythro-sphingosine,DMS)50μmol/L;Sphk1激活组加入佛波醇-12-豆蔻酸酯-13-乙酸酯(phorbol12-myristate-13-acetate,PMA)100nmol/L;对照组加入等量的培养基.采用MTT法检测细胞生长增殖;透射电镜观察细胞形态学变化;Matrigel三维培养法观察VM形成能力;Transwell小室模型观察细胞侵袭迁移能力的变化,QT-PCR、Western blot及ELISA技术检测血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达.结果:DMS显著抑制细胞的增殖、侵袭、迁移并促进细胞凋亡;透射电镜可见典型的凋亡特征;三维培养中不能形成管状VM;明显减弱VEGFmRNA及蛋白表达.PMA则显著促进HT-29细胞的增殖、侵袭、迁移并抑制细胞的凋亡;透射电镜可观察到细胞增殖特征;促进三维培养中管状VM的形成;明显增强VEGFmRNA及蛋白表达.对照组、DMS组及PMA组的侵袭细胞数:112.00±6.25vs57.00±8.00,142.00±5.57;迁移细胞数:69.33±4.04vs42.00±4.16,111.00±8.03;VEGFmRNA表达量:1.000vs0.740±0.122,1.220±0.075;VEGF蛋白表达:0.39±0.05vs0.23±0.02,0.65±0.06;VEGF蛋白分泌:103.00±8.96vs63.89±8.44,201.01±17.93,均P<0.05.结论:Sphk1可促进HT-29细胞增殖、侵袭、迁移并抑制细胞的凋亡,诱导VM的形成,其机制可能是通过增强结肠癌细胞的侵袭迁移能力并促进VEGF表达而发挥作用.AIM： To investigate the role of sphingosine kinase 1 （Sphkl） in vasculogenic mimicry （VM） formation in human colon cancer cell line HT±29 in vitro. METHODS： HT±29 cells were divided into three groups and treated with 100 nm/L of phorbol 12±myristate 13±acetate （PMA, Sphkl activa± tion group）, 50 mol/L of N,N±dimethyl±D± erythro±sphingosine （DMS, suppression group）, and equal volume of culture medium （control group）, respectively. After treatment, cell pro± liferation was determined by MTT assay, andcell invasiveness and migration were assessed by Transwell chamber assays. Cell apoptosis was observed by transmission electron micros± copy （TEM）. VM formation was observed in a three±dimensional culture system. The mRNA and protein expression of vascular endothelial growth factor （VEGF） was evaluated by QT±PCR and Western blot, respectively. The secretion of VEGF was detected by ELISA. RESULTS： Treatment with DMS significantly suppressed cell proliferation, invasion and mi± gration, promoted apoptosis, down±regulated VEGF mRNA and protein expression and se± cretion, and did not induce VM formation. In contrast, treatment with PMA significantly pro± moted cell proliferation, invasion （112.00 ± 6.25 vs 57.00 ± 8.00, 142.00 ± 5.57, both P 〈 0.05） and migration （69.33 ± 4.04 vs 42.00 ± 4.16, 111.00 ± 8.03, both P 〈 0.05）, suppressed apoptosis, up± regulated VEGF mRNA （1.000 vs 0.740 ± 0.122, 1.220 ± 0.075, both P 〈 0.05） and protein （0.39 ± 0.05 vs 0.23 ± 0.02, 0.65 ± 0.06, both P 〈 0.05） ex± pression and secretion （103.00 ± 8.96 vs 63.89 ± 8.44, 201.01 ± 17.93, both P 〈 0.05）, and induced the formation of tubular VM. CONCLUSION： Sphkl promotes cell prolifera± tion, invasion and migration, suppresses cell apoptosis, and induces VM formation possibly by up±regulating VEGF expression and secretion in human HT±29 colon cancer cell line.

关 键 词：血管生成拟态 鞘氨醇激酶-1 人结肠癌细胞 血管内皮生长因子 侵袭能力 

分 类 号：R735.35[医药卫生—肿瘤]