Screening and cloning of multi-drug resistant genes in HL-60/MDR cells.
OBJECTIVE
To screen and clone multi-drug resistance (MDR) related genes in MDR acute myeloid leukemia cells (HL-60/MDR).


METHODS
HL-60/MDR was established using All-Trans Retinoic Acid. With the HL-60 cells as "tester" and HL60/MDR as "driver", the cDNA library of HL-60/MDR was established by suppression subtractive hybridization. Then 12 of the resulting subtracted cDNA clones were selected for DNA sequencing and homology analysis. The obtained expressed sequence tags (ESTs) were analyzed with the GenBank BLASTN program to identify sequence homologies to known genes.


RESULTS
The HL-60/MDR cells had different multi-drug resistance to six kinds of chemotherapeutic drugs. The 211 positive gene clones in differential cDNA library of HL-60/MDR cells were amplified with PCR and 46 gene clones exhibited differential expression (ratio >3). Twelve gene clones with significant differential expression (ratio >5) were screened out to homology analysis. Of these, 11 matched known genes and the rest 1 showed no significant homology to human or non-human known sequences. It was named as gene clone HA117.


CONCLUSIONS
This effort provides the partial list of genes differential expressed in HL-60/MDR cells and a novel gene HA117 was found to be related to MDR. Identification of these genes contributes to our understanding of MDR development, and potentially provides candidate target genes to overcome MDR.
Zheng
Gai-Huan
GH
Affiliated Children's Hospital, Chongqing Medical University, Chongqing, China.
Fu
Jin-Rong
JR
Xu
You-Hua
YH
Jin
Xian-Qing
XQ
Liu
Wen-Li
WL
Zhou
Jian-Feng
JF
D000970:Antineoplastic Agents / Q000494:pharmacology; D003001:Cloning, Molecular; D018432:Drug Resistance, Multiple / Q000187:drug effects / Q000235:genetics*; D019008:Drug Resistance, Neoplasm / Q000187:drug effects / Q000235:genetics*; D020224:Expressed Sequence Tags; D015723:Gene Library; D052138:Genes, Neoplasm; D018922:HL-60 Cells; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods; D014212:Tretinoin / Q000494:pharmacology
D000970:Antineoplastic Agents; D014212:Tretinoin