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目的 确定非磷脂脂质体 (non phospholipidliposome ,NPL)介导反义c myc基因转染人脑胶质瘤 (glioma)的作用。方法 以金属离子诱导表达人反义c myc基因的质粒pHMTASCMHDNA与NPL结合转染人脑恶性胶质瘤细胞U2 5 1,转染 6h后加入 2 0 0 μmol/LZnCl2 作实验组 ,不加ZnCl2 转染细胞和单用NPL为对照组。在实验的第 2 4、48、72和 96h分别测定细胞还原四甲基偶氮唑盐 (MTT)的光吸收值。将实验第 2 4小时或 72小时的细胞分别进行annexin V FLUOS或TUNEL染色 ;细胞裂解后做ELISA检测c myc基因表达产物含量的变化。结果 显微镜下观察转染 48h实验组瘤细胞开始出现多角型细胞 ,细胞胞浆内产生空泡 ;MTT实验结果发现其细胞活性下降 ;annexin V FLUOS染色与TUNEL实验显示细胞出现明显的程序性细胞死亡特征 ;c myc基因表达产物下降。结论 非磷脂脂质体介导反义c myc基因转染脑胶质瘤细胞可特异性地抑制c myc基因表达 ,抑制瘤细胞生长 ,细胞活性下降 ,乃至出现程序性细胞死亡
Objective To determine the effect of nonphospholipid liposome (NPL) on the transfection of human glioma with antisense c myc gene. Methods Human glioma cell line U2 5 1 was transfected with pHMTASCMH DNA, a plasmid expressing human antisense c myc gene induced by metal ions. After transfection for 6 h, 200 micromol / L ZnCl2 was added to the experimental group without ZnCl2 Dyed cells and NPL alone were used as control. The light absorbency values of cell reduction tetramethylazolesalt (MTT) were determined at the 2nd, 4th, 48th, 72nd and 96h after the experiment. The cells at the 24th hour or 72th hour after the experiment were stained with annexin V FLUOS or TUNEL, respectively. After the cells were lysed, the expression of cmyc gene was detected by ELISA. Results Under the microscope, the polygonal cells began to appear in the tumor cells 48h after transfection, and the vacuoles were found in the cytoplasm of the transfected cells. MTT assay showed that the cell activity was decreased. Annexin V FLUOS staining and TUNEL assay showed that the cells showed obvious programmed cell death Features; c myc gene expression products decreased. Conclusion Non-phospholipid-mediated liposome-mediated antisense c myc gene transfected into glioma cells can specifically inhibit c-myc gene expression, inhibit tumor cell growth, decreased cell activity, and even apoptosis