非磷脂脂质体介导反义c-myc基因质粒转染对人脑胶质瘤细胞U251的影响  

Transfection of antisense c-myc into human glioma U251 cells by dint of non-phospholipid liposome

作　　者：朱俊铭[1] 全家妩[1] 

机构地区：[1]武汉生物制品研究所肿瘤研究室,430060

出　　处：《中华微生物学和免疫学杂志》2002年第1期92-94,共3页Chinese Journal of Microbiology and Immunology

摘　　要：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 。Objective To explore the effect of antisense c-myc on human glioma cell U251 after transfection by dint of non-phospholipid liposome (NPL). Methods pHMTASCMH, which inducibly expresses human antisense c-myc under the control of human metallothionein promoter, was mixed with NPL and added into U251 cell culture. ZnCl 2 (200μmol/L) was added after 6 hours. 3-[4,5-dimethylthiazolzyl]-2,5-diphenyl tetrazolium bromide (MTT) test was used to evaluate cell activity after 24-96 hours of the antisense c-myc transfection. The cells treated for 24 or 72 hours were microscoped in fluoresce microscope after Annexin-V-FLUOS staining or TUNEL staining respectively. The treated cells were lysed and then c-myc protein expression was assayed by ELISA. Results Complex of antisense c-myc and NPL significantly inhibited the proliferation of glioma cells in the presence of Zn 2+. NPL did not significantly affect cell growth at the concentration tested. The treated cells lost normal morphology after 24 hours and cytoplasmic vacuoles were found. Phenomena correlative with programmed cell death were well-identified in treated cells after Annexin-V-FLUOS or TUNEL staining. ELISA demonstrated the decrease of c-myc expression products. Conclusion Antisense c-myc inhibited the expression of c-myc, and proliferation of glioma cells and induced cell death.

关 键 词：反义C-MYC 脑胶质瘤 非磷脂脂质体 基因表达 细胞凋亡 U251 免疫疗法 动物实验 

分 类 号：R739.41[医药卫生—肿瘤] R730.51[医药卫生—临床医学]