一株新的红白血病细胞系(HIE1)的建立及其生物学特性

Establishment and biological characteristics of a novel erythroleukemia cell line(HIE1)

目的 建立一株具有原代细胞遗传学标志的白血病细胞系 ,并初步鉴定其生物学特性。方法 液体培养法建立细胞系 ,用R显带和逆转录 聚合酶链反应方法检测其遗传学标志 ;瑞氏染色、超微结构及组织化学染色观察细胞形态 ;用单克隆抗体检测细胞表面抗原 ;氰化高铁法和血红蛋白电泳测定血红蛋白 ;用联苯胺细胞染色观察阿糖胞苷 (Ara C)诱导后的红系的分化 ;用细胞形态学和吞噬功能实验观察佛波酯 (PMA)诱导的单核 巨噬细胞的分化。结果 由 1例慢性粒细胞白血病 (CML)急粒变患者外周血细胞 ,用液体培养 ,经 6 0余次传代 ,克隆出一株仍具原代细胞标志 (Ph+ ,abl/bcr+ )的白血病细胞株 ,命名为HIE1。该细胞具有粒 单系和红系血型糖蛋白A表型。HIE1细胞含有血红蛋白 ,且电泳后可见HbA和HbA2 带。加入 3.6× 10 -4 mmol/LAra C诱导后 ,联苯胺染色阳性细胞增加 ;PMA诱导后 1/ 3细胞贴壁 ,并呈梭样改变 ,6 .5 %的细胞具吞噬作用。瑞氏染色可观察到两类细胞 :一类为淡蓝色胞浆 ,少量细胞含有嗜天青颗粒 ;另一类为深蓝色胞浆 ,多空泡和伪足 ,无颗粒 ;组织化学结果显示 :POX、SB、CE为阴性 ,而AE、PAS、ACP为阳性。该细胞集落形成率为 37% ,倍增时间为 2 2～2 4h ,且EB病毒检测为阳性。结论 建立了一株具有粒 单系?

Objective To establish a novel leukemia cell line and characterize its biological characteristics. Method The cell line was established by liquid cell culture. The genetic marker was analyzed by R banding and reverse transcriptase polymerase chain reaction (RT PCR),cell morphology by microscopy, electron microscopy and histochemical staining, cell surface antigen by monoclonal antibody, hemoglobin by hyperomethemoglobin measurment and electrophoresis, erythroid differentiation by benzidine staining, and monocyte macrophage differentiation by cell morphology and phagocytosis. Results A novel erythroleukemia cell line (HIE1), with original cell genetic marker (Ph chromosome, bcr/abl fusion gene rearrangement), was established from a CML patient in blast crisis, and has been passaged for over 60 generations. Myelomonocyte marker and hemoglycoprotein A were found on the cell surface. HIE1 cells contained hemoglobin, the same HbA and HbA 2 bands as in normal individuals were displayed by Hb electrophoresis. The benzidine positive HIE1 cells were induced after exposure to 3.6×10 -4 mmol/L Ara C. When HIE1 cells were treated with 100?ng/ml PMA for 3 days, one third of the cells became spindle in shape, and 6.5% of the cells exert phagocytosis. The cells were classified into two types with Wright staining: one showing light blue cytoplasm and a few of cells with basophilic granules, the other showing dark blue cytoplasm with vacuoles and pseudopods without granules. In addition, POX,SB, CE stains were negative, and AE,PAS, ACP stains positive. Colony formation of the cells was 37%, the cell doubling time was 22～24 hrs, and EB virus detection was positive. Conclusion A novel erythroleukemia cell line with bcr/abl fusion gene and characteristics of myelomonocytic and erythroid cells was established.