PML和PML-RARα反义寡聚核苷酸对急性早幼髓性白血病细胞NB4的影响  

Effects of PML and PML-RARα antisence oligonucleotides on promyelocytic leukemia cell line NB4

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

机构地区：[1]南通医学院附属医院血液科,226001 [2]上海第二医科大学附属仁济医院,200001

出　　处：《中华肿瘤杂志》2000年第4期283-286,共4页Chinese Journal of Oncology

基　　金：国家自然科学基金资助项目 !(395 90 2 91)

摘　　要：目的　比较抗早幼粒细胞白血病 (promyelocyticleukemia ,PML)基因反义核酸 (STAS)和抗PML RARα反义核酸 (FUAS)对NB4细胞生长、细胞分化、PML RARαmRNA表达及PML/PML RARα蛋白细胞内定位的影响。方法　以NB4细胞株为研究对象 ;采用台盼蓝排除计数板计数法、甲基纤维素半固体培养、RT PCR法、免疫荧光等技术进行研究。结果STAS组、FUAS组明显抑制NB4细胞增殖和白血病细胞集落 (AML CFU)形成 ;处理后第 5天可见细胞部分分化 ,FUAS组尤为明显 ;PML RARαmRNA表达 2 4h即明显减少 ,持续到 72h。抗PML抗体免疫荧光检测 ,STAS组 ,2 4h后细胞内颗粒明显减少 ,72h细胞内颗粒几乎完全消失 ;FUAS组 ,2 4h后细胞内微小颗粒消失 ,残存的颗粒变大 ,12 0h细胞内颗粒几乎消失。结论　STAS、FUAS均能阻断PML RARαmRNA表达和蛋白质翻译 ,促进细胞分化 ;Objective To investigate the different effects of anti PML (promyelocytic leukemia) and anti PML/RARα (promyelocytic leukemia/retionic acid receptor α) antisense oligonucleotides on cell growth, expression of PML RARα mRNA and PML RARα/PML protein location of NB4 cell line. Methods RT PCR was used for PML RARα mRNA expression, trypan blue exclusion for cell count, methylcellulose assay for leukemic colony forming unit, immuno fluorescence for PML RARα/PML protein localization. Results Both anti PML start codon region antisence (STAS) and anti PML RARα fusion region antisence (FUAS) could inhibit cell growth and formation AML CFU. Cells became partially differentiated on day 5, being more marked in FUAS treated cells than in STAS treated ones. Down regulated PML RARα mRNA expression occurred at 24 h was in STAS and FUAS treated cells and maintained for up to 72 h. Immuno fluorescence analysis with anti PML monoclonal antibody showed a remarkable decrease to almost complete disappearance of microgranules. The residual granules became enlarged to become discrete dots (<10 per cell), similar to normal POD structure in some STAS treated cells at 24 h. At 72 h, nearly all the granules disappeared. Similar changes were observed in FUAS treated cells. Conclusion Both PML and PML RARα antisense oligonucleotides can specifically block the expression of PML RARα at mRNA and protein levels. PML protein is implicated in the regulation of cell differentiation.

关 键 词：白血病 APL PML-RARα基因 反义寡核苷酸 

分 类 号：R733.7[医药卫生—肿瘤]