抗PML-RAR_α反义核酸对NB4细胞形态、PML-RAR_α mRNA及蛋白质表达的影响  

Effects of antiPMLRAR antisense on cell morphology and expression of PMLRAR mRNA and protein of NB4 cells

作　　者：陈烨[1,2] 缪金明[1,2] 朱学宏[1,2] 邵念贤[1,2] 方智雯[1,2] 欧阳仁荣[1,2] 

机构地区：[1]上海第二医科大学仁济医院血液科 [2]上海血液学研究所

出　　处：《中华血液学杂志》1999年第6期307-309,共3页Chinese Journal of Hematology

摘　　要：目的探讨抗ＰＭＬＲＡＲα反义核酸（ＦＵＡＳ）对ＮＢ４细胞的分化、ＰＭＬＲＡＲαｍＲＮＡ表达及ＰＭＬ／ＰＭＬＲＡＲα蛋白细胞内定位的影响。方法以ＮＢ４细胞为研究对象，细胞形态观察采用Ｗｒｉｇｈｔ染色法；ＰＭＬＲＡＲαｍＲＮＡ表达采用ＲＴＰＣＲ法；ＰＭＬ／ＰＭＬＲＡＲα蛋白细胞内定位采用免疫荧光技术。结果ＦＵＡＳ处理后第５天，细胞出现部分分化。处理后２４，７２，１２０小时，ＰＭＬＲＡＲαｍＲＮＡ表达分别下降５２．０％、６８．７％、２３．４％。抗ＰＭＬ抗体免疫荧光检测，ＦＵＡＳ处理２４小时后，细胞内微小颗粒消失，残存颗粒变大，１２０小时细胞内颗粒几乎消失。结论ＦＵＡＳ特异性阻断ＰＭＬＲＡＲαｍＲＮＡ表达，使ＰＭＬＲＡＲα融合蛋白合成减少甚至消失。Objective To investigate the effects of antiPMLRAR antisense (FUAS) on cell morphology, expression of PMLRAR mRNA and PMLRAR/PML protein localization of NB4 cells. Methods PMLRAR mRNA expression was assayed by RTPCR and PMLRAR/PML protein localization by immunofluorescence. Results NB4 cells were partially differentiated after 5 days of FUAS treatment and typical apoptosis was found after 7 days incubation with FUAS. The expression of PMLRAR mRNA at 24h was already down regulated in FUAStreated cells. After 24h, 72h and 120h incubation with FUAS, PMLRAR mRNA showed 52.0%, 68.7% and 23.4% reductions, respectively, as compared with that of control. Immunofluorescence analysis with antiPML monoclonal antibody showed disappearance of microgranules, residual granules becoming larger discrete dots at 24h of FUAS treatment and almost disappearence of dots at 120h. Z]Conclusion FUAS specifically blocks the expression and translation of PMLRAR gene, makes the production of PMLRAR fusion protein decrease or disappear and prompts cell differentiation.

关 键 词：PML-RARΑ NB4细胞系 白血病 反义核酸 MRNA 

分 类 号：R733.71[医药卫生—肿瘤]