Beran M, Pisa P, O'Brien S, Kurzrock R, Siciliano M, Cork A, Andersson B S, Kohli V, Kantarjian H
Department of Hematology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Cancer Res. 1993 Aug 1;53(15):3603-10.
The establishment and the biological properties of a new leukemic cell line (KBM-5) derived from a patient in the blastic phase of chronic myelogenous leukemia are described. The cells exhibited multiple copies of the Philadelphia chromosome, and a high level of p210Bcr-Abl kinase activity was detected with rabbit anti-Abl and anti-Bcr (exon 3) peptide antisera. Use of specific primers and polymerase chain reaction followed by Southern blotting revealed that KBM-5 cells carried a bcr3-ABLII splice junction. While a normal BCR message was detected, no normal ABL message was found. The cells were phenotypically myeloid with monocytic differentiation. The high cloning efficiency in semisolid media was independent of the presence of exogenous colony-stimulating factors. In vitro exposure to induces of differentiation, such as retinoic acid, dimethyl sulfoxide, or hemin, failed to influence the growth rate of the cells and their level of differentiation. KBM-5 cells are highly resistant to the antiproliferative action of recombinant alpha- and gamma-interferons. Although sensitive to recombinant tumor necrosis factor alpha, they were completely resistant to natural killer cell action. KBM-5 cells constitutively expressed mRNA for tumor necrosis factor alpha but not for gamma-interferon, other interleukins, or hematopoietic growth factors. The KBM-5 cells that were transplanted into SCID mice manifested metastatic potential and tissue invasiveness similar to the way leukemic cells in humans do. This new KBM-5 cell line represents a helpful model for examining in vitro and in vivo modulation of the growth and properties of leukemic cells by using biological and chemotherapeutic agents.
本文描述了从一名慢性粒细胞白血病急变期患者中分离出的一种新的白血病细胞系（KBM-5）的建立及其生物学特性。这些细胞呈现出多个费城染色体拷贝，用兔抗Abl和抗Bcr（外显子3）肽抗血清检测到高水平的p210Bcr-Abl激酶活性。使用特异性引物和聚合酶链反应，随后进行Southern印迹分析，结果显示KBM-5细胞携带bcr3-ABLII剪接连接。虽然检测到正常的BCR信息，但未发现正常的ABL信息。这些细胞在表型上呈髓系，具有单核细胞分化特征。在半固体培养基中具有较高的克隆效率，且与外源性集落刺激因子的存在无关。体外暴露于分化诱导剂，如维甲酸、二甲基亚砜或血红素，均未能影响细胞的生长速率及其分化水平。KBM-5细胞对重组α和γ干扰素的抗增殖作用具有高度抗性。虽然对重组肿瘤坏死因子α敏感，但它们对自然杀伤细胞的作用完全抗性。KBM-5细胞组成性表达肿瘤坏死因子α的mRNA，但不表达γ干扰素、其他白细胞介素或造血生长因子的mRNA。移植到SCID小鼠体内的KBM-5细胞表现出与人类白血病细胞相似的转移潜能和组织侵袭性。这种新的KBM-5细胞系为利用生物和化学治疗药物研究白血病细胞生长和特性的体外和体内调节提供了一个有用的模型。
