siRNA下调鞘氨醇激酶-1对结肠癌LOVO细胞凋亡和侵袭的影响及其机制的研究  

Effect of sphingosine kinase 1 siRNA transfection on colon cancer LOVO cell apoptosis and invasion and its molecular mechanisms

作　　者：刘诗权[1] 钟月圆[1] 黄杰安[1] 覃蒙斌[1] 唐国都[1] 姜海行[1] 

机构地区：[1]广西医科大学第一附属医院消化内科,广西南宁530021

出　　处：《实用肿瘤杂志》2011年第5期468-472,共5页Journal of Practical Oncology

基　　金：国家自然科学基金项目(30760275);广西科技厅留学回国基金项目(桂科回0832008);广西卫生厅基金项目(Z2008107)

摘　　要：目的研究siRNA下调鞘氨醇激酶-1(Sphk1)的表达对结肠癌LOVO细胞增殖、凋亡、侵袭以及ERK和p38通路的影响。方法采用siRNA抑制Sphk1的表达;采用MTT法测定细胞的增殖,流式细胞术分析细胞的凋亡,Transwell小室检测细胞侵袭力的变化,Western杂交检测蛋白的表达;采用RT-PCR法检测细胞Sphk1 mRNA表达,ELISA法检测细胞基质金属蛋白酶(MMP)-2、MMP-9和尿激酶纤维蛋白溶酶原激活剂(uPA)的分泌。结果 Sphk1 siRNA显著抑制Sphk1 mRNA和蛋白表达,并可诱导细胞的凋亡,抑制细胞的增殖和侵袭。抑制Sphk1的表达可降低ERK和磷酸化ERK蛋白的表达,并促进p38和磷酸化p38蛋白的表达,同时可抑制细胞MMP-2、MMP-9和uPA的分泌。结论抑制Sphk1可能是通过抑制ERK并激活p38 MAPK通路,下调MMP-2、MMP-9和uPA的分泌,从而抑制结肠癌细胞的增殖和侵袭并促进细胞的凋亡。Objective To investigate the effect of sphingosine kinase 1（Sphk1） siRNA transfection on the biological features of colon cancer LOVO cell and to explore its molecular mechanisms. Methods Sphk1 siRNA was exploited to suppress the expression of Sphk1.Cell proliferation was detected by the method of MTT and cell apoptosis was examined by flow cytometry.Invasion capabilities of cells were assessed by the transwell chambers model.Western blotting was exploited to evaluate the protein expression and RT-PCR was used to detected the mRNA expression.Concentrations of MMP-2,MMP-9 and uPA were detected by the method of ELISA. Results Sphk1 siRNA transfection significantly suppressed the mRNA and protein expression of Sphk1.Suppression of Sphk1 suppressed the cell proliferation and invasion,meanwhile promoted the cell apoptosis.With the down regulation of Sphk1,the protein expressions of ERK,p-ERK and the secretion of MMP-2,MMP-9,uPA were strikingly suppressed,inversely the protein expression of p38 and p-p38 were enhanced. Conclusions Suppression of Sphk1 potently suppresses colon cancer LOVO cell proliferation and invasion,meanwhile enhances cell apoptosis.Reduction of the secretion of MMP-2,MMP-9 and uPA via suppressing ERK and activating p38 pathway may be one of its molecular mechanisms.

关 键 词：结肠肿瘤 鞘氨醇激酶-1 细胞凋亡 肿瘤侵润 肿瘤细胞 培养的 

分 类 号：R735.35[医药卫生—肿瘤]