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Cytogenetic and molecular analysis of the acute monocytic leukemia cell line THP‐1 with an MLL‐AF9 translocation
Author(s) -
Odero Maria D.,
ZeleznikLe Nancy J.,
Chinwalla Vandana,
Rowley Janet D.
Publication year - 2000
Publication title -
genes, chromosomes and cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.754
H-Index - 119
eISSN - 1098-2264
pISSN - 1045-2257
DOI - 10.1002/1098-2264(2000)9999:9999<::aid-gcc1040>3.0.co;2-z
Subject(s) - chromosomal translocation , karyotype , thp1 cell line , fluorescence in situ hybridization , acute monocytic leukemia , derivative chromosome , biology , monocytic leukemia , microbiology and biotechnology , leukemia , cytogenetics , molecular cytogenetics , chromosome , genetics , cell culture , gene
Cell lines derived from patients with leukemia are used in many molecular biology studies. Here we report the cytogenetic analysis of the THP‐1 cell line using G‐banding, fluorescence in situ hybridization (FISH), and spectral karyotyping (SKY), and the molecular characterization of the MLL‐AF9 rearrangement by RT‐PCR. The THP‐1 cell line was established from the peripheral blood of a 1‐year‐old boy with acute monocytic leukemia (AML‐M5). THP‐1 is near‐diploid and consists of two related subclones with a number of aberrations, including the t(9;11), associated with AML M5. The use of FISH allowed us to identify and characterize otherwise hidden cytogenetic rearrangements, which include duplication of the 3′ portion of MLL in the derivative 9 chromosome and a deletion of the 5′ portion of the AF9 gene involved in the translocation. In addition to confirming the FISH results, SKY allowed for a more precise characterization of the karyotype of THP‐1 and allowed us to identify other abnormalities in this cell line, including der(1)t(1;12), der(20)t(1;20), deletions 6p, 12p, and 17p, trisomy 8, and monosomy 10. Sequencing of the RT‐PCR product showed a direct in‐frame fusion product on the derivative chromosome 11 between exon 6 (exon 9) of MLL and exon 5 of AF9, which is most commonly involved in MLL‐AF9 translocations. This study demonstrates that combining different techniques to achieve a more precise characterization of the THP‐1 cell line provides important information that will be valuable for understanding the critical events required for leukemogenesis. © 2000 Wiley‐Liss, Inc.