p53 and N-ras mutations in two new leukemia cell lines established from a patient with multilineage CD7-positive acute leukemia

Abstract

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.

Near-triploidy and near-tetraploidy in hematological malignancies and mutation of the p53 gene

“…Genomic DNA was extracted from cell specimens using standard proteinase K digestion and phenol/chloroform extraction. DNA was subjected to polymerase chain reaction–single‐strand conformation polymorphism (PCR–SSCP) analysis to search for mutations in exons 5–8 of the p53 gene, as described previously (Abo et al. , 1993).…”

MDR1 mRNA Expressions in Peripheral Blood Mononuclear Cells of Patients with Ulcerative Colitis in Relation to Glucocorticoid Administration

“…From this point of view, the amount of P‐gp expressed on immune cells might also influence the therapeutic efficacy of GCs, which are known as substrates of P‐gp 19 . Overexpression of functional P‐gp or MDR1 mRNA has been reported to be induced by mutation of a tumor suppressor p53 gene 20 or by stimulation of cells by inflammatory chemical mediators such as TNFα 21,22 . In addition to these tissue or cellular factors, induction of P‐gp or MDR1 mRNA in cancer cells by several anticancer agents has been suggested 16–18 .…”

Establishment of a near‐triploid human B‐cell lymphoma cell line with t(14;18) and a p53 gene point mutation

“…Sequence analysis of the p53 gene. RT-PCR was performed as described elsewhere (Abo et al, 1993). The sense primers were ST1, SN2 and SC3.…”

“…The sense primers were ST1, SN2 and SC3. The antisense primers were AST1, ASN2 and ASC3 (Abo et al, 1993).…”

“…Cytogenetic analysis was performed after culturing the cells for 24–48 h. 20 metaphase TK cells were analysed after Trypsin‐Giemsa banding (Abo et al , 1993).…”