Abo J, Inokuchi K, Dan K, Nomura T
Third Department of Internal Medicine, Nippon Medical School, Tokyo, Japan.
Blood. 1993 Nov 1;82(9):2829-36.
Two new myeloid cell lines (K051 and K052) were established from a patient with multilineage CD7-positive acute leukemia. The K051 and K052 were established from the patient's bone marrow cells at diagnosis and at relapse, respectively. The K051 cell expressed myeloid-associated antigens (CD13 and CD33), a platelet-associated antigen (CD41), and an erythroid antigen (glycophorin A). The K052 cell expressed myeloid-associated antigens (CD13, CD14, and CD33), lymphoid markers (CD2, CD5, and CD7), and HLA-DR. Chromosome analysis of both cell lines showed a 17p- chromosome. Both cell lines were investigated for aberrations of the p53 gene and the N-ras gene. A p53 mutation detected in both cell lines consisted of a C-->T substitution in codon 248. An N-ras mutation detected only in the K052 cell consisted of a G-->C substitution in codon 13. Expression of the multidrug resistance gene (MDR1) was also investigated by the semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). MDR1-mRNA was more highly expressed by the K052 cell than the K051 cell, being equivalent to that in HEL cells. The functional MDR1-protein against vincristine was also observed, and its function was inhibited by verapamile and Cyclosporin A. The K052 cells were capable of phenotypic or morphologic differentiation after being incubated with granulocyte colony-stimulating factor, interleukin-2, phorbol 12-myristate 13-acetate, or 1,25-dihydroxy-vitamin D3. In contrast, the K051 cells responded phenotypically to retinoic acid. Thus, the K051 and K052 cell lines will be useful for investigating the cellular and molecular events in leukemogenesis and differentiation, and the mechanism of expression of the MDR1 gene.
从一名多谱系CD7阳性急性白血病患者中建立了两个新的髓系细胞系（K051和K052）。K051和K052分别从患者诊断时和复发时的骨髓细胞中建立。K051细胞表达髓系相关抗原（CD13和CD33）、血小板相关抗原（CD41）和红系抗原（血型糖蛋白A）。K052细胞表达髓系相关抗原（CD13、CD14和CD33）、淋巴样标志物（CD2、CD5和CD7）以及HLA-DR。两个细胞系的染色体分析均显示17号染色体短臂缺失。对两个细胞系均进行了p53基因和N-ras基因畸变的研究。在两个细胞系中检测到的p53突变均为密码子248处的C→T替换。仅在K052细胞中检测到的N-ras突变是密码子13处的G→C替换。还通过半定量逆转录聚合酶链反应（RT-PCR）研究了多药耐药基因（MDR1）的表达。K052细胞中MDR1-mRNA的表达高于K051细胞，与HEL细胞中的表达水平相当。还观察到了针对长春新碱的功能性MDR1蛋白，其功能可被维拉帕米和环孢素A抑制。用粒细胞集落刺激因子、白细胞介素-2、佛波醇12-肉豆蔻酸酯13-乙酸酯或1,25-二羟基维生素D3孵育后，K052细胞能够进行表型或形态学分化。相比之下，K051细胞对维甲酸有表型反应。因此，K051和K052细胞系将有助于研究白血病发生和分化过程中的细胞和分子事件以及MDR1基因的表达机制。
