摘要
目的探讨抗PMLRARα反义核酸(FUAS)对NB4细胞的分化、PMLRARαmRNA表达及PML/PMLRARα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;PMLRARαmRNA表达采用RTPCR法;PML/PMLRARα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,PMLRARαmRNA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PMLRARαmRNA表达,使PMLRARα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.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
1999年第6期307-309,共3页
Chinese Journal of Hematology