抗PML—RARα反义核酸对NB4细胞形态,PML—RARα mRNA及蛋白质？…

目的 探讨抗ＰＭＬ－ＲＡＲα反义核酸（ＦＵＡＳ）对ＮＢ４细胞的分化、ＰＭＬ－ＲＡＲα ｍＲＮＡ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染以法；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－

Effects of anti-PML-RAR alpha antisense on cell morphology and expression of PML-RAR alpha mRNA and protein of NB4 cells]

OBJECTIVE: To investigate the effects of anti-PML-RAR alpha antisense (FUAS) on cell morphology, expression of PML-RAR alpha mRNA and PML-RAR alpha/PML protein localization of NB4 cells. METHODS: PML-RAR alpha mRNA expression was assayed by RT-PCR and PML-RAR alpha/PML protein localization by immuno-fluorescence. 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 PML-RAR alpha mRNA at 24 h was already down regulated in FUAS-treated cells. After 24 h, 72 h and 120 h incubation with FUAS, PML-RAR alpha mRNA showed 52.0%, 68.7% and 23.4% reductions, respectively, as compared with that of control. Immuno-fluorescence analysis with anti-PML monoclonal antibody showed disappearance of microgranules, residual granules becoming larger discrete dots at 24 h of FUAS treatment and almost disappearence of dots at 120 h. CONCLUSION: FUAS specifically blocks the expression and translation of PML-RAR alpha gene, makes the production of PML-RAR alpha fusion protein decrease or disappear and prompts cell differentiation.