ING4蛋白对人胶质瘤U251细胞增殖及凋亡的影响

Effect of ING4 protein on proliferation and apoptosis in human U251 glioma cells

目的探讨ING4(inhibitor of growth family 4)蛋白对人胶质瘤U251细胞增殖及凋亡的影响。方法用PEI转染ING4蛋白过表达(pEGFP-C2/ING4转染组)及阴性对照载体(pEGFP-C2转染组)至人胶质瘤U251细胞,G418筛选后建立ING4蛋白稳定表达细胞株;RT-PCR、免疫组化、Western blot检测ING4蛋白的表达,MTT法和Hochest法检测U251细胞的增殖和凋亡。结果 pEGFP-C2/ING4转染组和pEGFP-C2转染组转染U251细胞效率分别为84%和82%;RT-PCR、免疫组化、Western blot均检测到pEGFP-ING4转染组ING4的强表达;与pEGFP-C2转染组和空白对照组相比,pEGFP-C2/ING4转染组的U251细胞72 h后生长抑制率和凋亡率有统计学差异(P<0.05)。结论 ING4蛋白对胶质瘤U251细胞有显著的增殖抑制和诱导凋亡作用。

Objective To investigated the effect of inhibitor of growth family 4（ING4） protein on proliferation and apoptosis in human U251 glioma cells.Methods Human U251 glioma cells were transfected with plasmid pEGFP-C2/ING4（ING4 protein group） and plasmid pEGFP-C2（pEGFP-C2 group） via PEI,respectively,and the non-transfected U251 cells were used as blank control（blank group）.Positive cells were screened by G418 to establish a cell strain with stable expression of ING4 protein.ING4 protein expression was detected by RT-PCR,immunohistochemistry and Western blotting,while proliferation and apoptosis of transfected and non-transfected human U251 glioma cells were detected by MTT assay and Hochest staining.Results The efficiency of pEGFP-C2/ING4 and pEGFP-C2 tranfection into human U251 glioma cells were 84% and 82%,respectively.RT-PCR,immunohistochemistry and Western blotting showed significant up-regulation of ING4.Compared with the pEGFP-C2 group and blank group,the growth inhibition rate and apoptotic rate of human U251 glioma cells after 72 h transfection in the pEGFP-C2/ING4 group were significantly higher（P〈0.05）.Conclusion ING4 protein can significantly inhibit proliferation and induce apoptosis in human U251 glioma cells.