[In silicon cloning of HV126, a novel human gene related to multi drug resistance in leukemia].
OBJECTIVE
To find the novel gene related to the multi-drug resistance in leukemia and explore the molecular mechanism of multi-drug resistance.


METHODS
The subtracted HL-60/VCR cDNA library was generated through the suppression subtractive hybridization using the wild HL-60 cells' cDNA as target and HL-60/ ATRA cells' as driver. A novel expression sequence tag (EST) sequence, which differentially expressed in HL-60/ ATRA cell, was screened by cDNA chip. Then a novel human gene, HV126 was assembled by the EST assembly tools. Bioinformatical databases and softwares were used to analyze and predict its function. Reverse transcription-PCR (RT-PCR) was used to detect the expression of HV-126 gene in leukemia cells before and after chemotherapy.


RESULTS
The full open reading frames (ORFs) of the novel EST assembled by overlapping dbEST sequences included a 1991 bp nucleic sequence, which was named HV126. The deduced amino acid sequence consisted of 365 amino acids. The sequence of the novel gene exhibited 43% homology to a known gene, which is a possible member of the death domain-flood family implicated in apoptosis and inflammation. The expression of HV126 was proved to be related to the drug sensitivity in leukemia cells by RT-PCR.


CONCLUSIONS
HV126, the novel gene, might have roles in regulating multi-drug resistance in leukemia. Further laboratory research should be done on cloning and making clear the gene function.
Fu
Jinrong
J
Department of Hematology, Tongji Hospital,Tongji Medical College, Huazhong University of Science and Technology, Wuhanì Hubei, 430030 P. R. China. jrfu@tjh.tjmu.edu.cn
Liu
Wenli
W
Zhou
Jianfeng
J
Zhou
Yufeng
Y
Xu
Youhua
Y
Jin
Xianqing
X
D000970:Antineoplastic Agents / Q000494:pharmacology; D001483:Base Sequence; D002874:Chromosome Mapping; D002883:Chromosomes, Human, Pair 14 / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D018432:Drug Resistance, Multiple / Q000235:genetics*; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects; D018922:HL-60 Cells; D006801:Humans; D007938:Leukemia / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008969:Molecular Sequence Data; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D000970:Antineoplastic Agents; D018076:DNA, Complementary; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C539382:HV126 protein, human