EFFECTS OF ANTI-PML/RARα OR ANTI-PML ANTISENCE ON THE GROWTH, DIFFERENTIATION AND APOPTOSISOF NB4 CELL LINES

Objectire To investigate the ellects of anti - PML/RARx or anti - PML antisence on the growth,dtherentiation and apoptosis of NB4 cell lines. Methods Wright’s stain for cell morphology, flow cytometry andDNA gel electronphoresis for cell apoptosis, methylcellulose assays for leukemic colony forming unit andtrypan - blue exclusion for cell counts. Results Both the start cordon region of the PML or PML - RARx mRNA(STAS) and the fusion point region of the long type PML - RARx mRNA (FUAS) could inhibit cell growth. Cellsbecame partially differentiated at 5d of treatment, and FUAS - treated cells showed more significant differentiationthan STAS- treated cells. Morphology of typical apoptosis could be seen at 7, 9d incubation with antisenceoligodeoxynucleotides (AS). In contrast, no cell growth inhibition, no morphology changes were seen in Sen or Rantreated cells compared with untreated cells. The number of acute myelocytic leukemia colony forming unit(AML - CFU) markedly decreased in STAS and FUAS treated cells. Cell DNA content analyzed by flow cytometryshowed the typical profile of apoptotic cells, in which pre - G1 peak appear before G1 peak at 7,9d of treatment withSTAS or FUAS. Conclusion Anti - PML/RARx or anti- PML antisence inhibit the cell growth, inducedifferentiation and differentiated cell apoptosis of NB4 cells.

抗PML-RARα反义核酸对NB4细胞生长、细胞周期及细胞凋亡的影响

目的：探讨抗ＰＭＬ－ＲＡＲα反义核酸对早幼粒白血病细胞生长、细胞周期、细胞凋亡的作用。方法：以ＮＢ４细胞株为研究对象；细胞计数采用台盼兰排除、计数板计数法；白血病细胞集落采用甲基纤维素半固体培养；流式细胞仪（ＦＣＭ）用于检测细胞凋亡和细胞周期。结果：以６０μｇ／ｍｌ的终浓度处理细胞，抗 ＰＭＬ－ＲＡＲα融合部位反义核酸（ＦＵＡＳ）能明显抑制ＮＢ４细胞增殖及白血病细胞集落（ＡＭＬ－ＣＦＵ）形成，最大抑制率分别为４６．８％，５１．６％；ＧＭ－ＣＳＦ能减弱ＦＵＡＳ的作用，ＦＵＡＳ作用时间越短，ＧＭ－ＣＳＦ促ＡＭＬ－ＣＦＵ增加越明显；ＦＵＡＳ处理第７，９天，出现明显的凋亡细胞群，凋亡细胞百分比分别为４１．０％，３４．２％；ＦＵＡＳ处理第５天Ｓ期下降，Ｇ２／Ｍ期升高，第７天Ｃ０／Ｇ１期明显升高，Ｓ期回升。结论：抗ＰＭＬ－ＲＡＲα反义核酸具有抑制早幼粒白血病细胞增殖、诱导细胞凋亡的作用。