NALP1基因与骨肉瘤细胞的相关研究

Research on NALP1 Gene and Osteosarcoma Cell

目的:探讨NALP1基因在骨肉瘤细胞株MG-63、U-2OS中的表达,以及高表达NALP1基因对于骨肉瘤细胞体外凋亡的影响。方法:使用RT-PCR、Western-blot法检测骨肉瘤细胞株MG-63、U-2OS中的mRNA及蛋白表达水平并与人成骨细胞株hFOB1.19比较。将NALP1基因转染质粒PcDNA3.1,将重组质粒转染骨肉瘤细胞,分成高表达基因组、空质粒组及对照组,加入抗肿瘤药物顺铂及甲氨蝶呤促使肿瘤细胞凋亡,使用流式细胞仪测定各组肿瘤细胞凋亡率。结果:通过统计分析,骨肉瘤细胞株MG-63、U-2OS中的mRNA及蛋白表达水平均低于人成骨细胞株hFOB1.19(P<0.05),NALP1高表达组的肿瘤细胞凋亡率明显高于空白质粒组及对照组。结论:上调骨肉瘤细胞株MG-63、U-2OS中的NALP1的表达量可以促进肿瘤细胞凋亡。

Objective： To investigate the expression of NALP1 gene in osteosarcoma cell line MG-63,U-2OS and effect in vitro apoptosis about high expression of NALP1 gene for osteosarcoma cells.Methods： We use the methods of RT-PCR,Western-blot to assay the expression of mRNA and protein in osteosarcoma cell line MG-63,U-2OS and compare with human osteoblast cell line hFOB1.19.We transfected the NALP1 gene into plasmid PcDNA3.1,and the recombinant plasmid was.We divided the transfected osteosarcoma cells into three groups,highly expressed genes group,blank plasmid group and control group.Then we add the anticancer drugs cisplatin and methotrexate to promote tumor cell apoptosis.At last we use flow cytometry in each group to determine the apoptosis of tumor cell.Results： Through statistical analysis,we get the result that mRNA and protein expression in osteosarcoma cell line MG-63,U-2OS were lower than the human osteoblast cell line hFOB1.19（P 0.05）,and apoptosis of highly expressed NALP1 gene group was significantly higher than the blank plasmid group and the control group.Conclusions： Increasing the expression of NALP1 gene in osteosarcoma cell line MG-63 and U-2OS can promote apoptosis of tumor cell.