摘要
目的通过抑制骨肉瘤细胞株MG63中FoxM1基因表达,观察对骨肉瘤细胞系增殖与凋亡的影响。方法采用蛋白质印迹法检测不同人骨肉瘤细胞株MG63和SOSP-9607中FoxM1基因的表达。将骨肉瘤细胞株MG63分为实验组、阴性对照组和空白对照组,实验组转染FoxM1特异性siRNA(FoxM1-siRNA),阴性对照组转染阴性对照siRNA,空白对照组不做处理。实时-聚合酶链反应(Real-time PCR)和蛋白质印迹法检测转染前后FoxM1基因在mRNA及蛋白表达水平的变化;四甲基偶氮唑盐(MTT)比色法检测细胞增殖率,流式细胞仪测定细胞凋亡率。结果人骨肉瘤细胞株MG63和SOSP-9607中存在FoxM1基因的表达。实验组MG63细胞转染siRNA后,FoxM1的mRNA表达为0.39±0.09,低于阴性对照组的0.91±0.07,t=7.92,P<0.001;也低于空白对照组的0.96±0.08,t=8.68,P<0.001。实验组FoxM1蛋白表达为0.31±0.06,显著低于阴性对照组的0.81±0.10,t=7.20,P<0.001;也低于空白对照组的0.89±0.09,t=8.35,P<0.001。实验组MG63细胞增殖率为0.36±0.03,显著低于阴性对照组的0.61±0.02,t=12.86,P<0.001;也低于空白对照组的0.65±0.02,t=14.92,P<0.001。实验组MG63细胞凋亡率为(13.69±1.15)%,显著高于阴性对照组的(2.38±0.90)%,t=14.16,P<0.001;也高于空白对照组的(2.87±0.86)%,t=13.54,P<0.001。结论人骨肉瘤细胞株中存在FoxM1蛋白的表达;特异性siRNA可下调骨肉瘤细胞株MG63中FoxM1基因的表达,对细胞的增殖具有负性调控作用,并能诱导细胞凋亡。
OBJECTIVE To detect the expression of FoxM1 in the human osteosarcoma cell line MG63 and SOSP-9607,and to investigate the influence of suppression of the expression of FoxM1 on proliferation and apoptosis in human osteosarcoma cell line MG63 and explore the role FoxM1 played in the pathogenesis of osteosarcoma.METHODSThe expression of FoxM1 was detected by Western-blot in osteosarcoma cell line MG63 and SOSP-9607.MG63 cells were divided into three groups：experimental group,negative control group and blank group.FoxM1-targeted siRNA and control siRNA were transfected into MG63 cells in experimental group and negative control group,respectively.Real-time reverse transcription-polymerase chain reaction（RT-PCR）and Western blot were used to detect the expression of mRNA and protein of FoxM1.MTT assay was used to detect the proliferation of MG63 cells and apoptosis was measured by flow cytometry.RESULTS The results of Western-blot showed that the expression of FoxM1 was detected in the human osteosarcoma cell line MG63 and SOSP-9607.FoxM1-targeted siRNA was successfully transfected into MG63 cells,leading to the significant inhibition of FoxM1 gene expression at both mRNA（0.39±0.09）and protein level（0.31±0.06）compared with negative control group（0.91±0.07,0.81±0.10）and blank group（0.96±0.08,0.89±0.09）,P0.001 for each.The MTT assay showed that FoxM1-targeted siRNA inhibited the proliferation of MG63 cells in experimental group（0.36±0.03）compared with two control groups（0.61±0.02,0.65±0.02,P0.001 for each）.Results of FCM pointed that the apoptosis ratio of experimental group（13.69±1.15）was significant higher than that of negative control group（2.38±0.90）and blank group（2.87±0.86,P0.001 for each）.CONCLUSIONS In vitro,FoxM1-targeted siRNA could effectively inhibit the expression of FoxM1 at both mRNA and protein level in human osteosarcoma cell line MG63.Downregulation of FoxM1 expression could inhibit cell proliferation of MG63 cells and induce cell apoptosis.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2014年第20期1575-1579,共5页
Chinese Journal of Cancer Prevention and Treatment
基金
国家自然科学基金(81072194)
