In silicon cloning of HV126, a novel human gene related to multi drug resistance in leukemia.
- Author:
Jinrong FU
1
;
Wenli LIU
;
Jianfeng ZHOU
;
Yufeng ZHOU
;
Youhua XU
;
Xianqing JIN
Author Information
- Publication Type:Journal Article
- MeSH: Antineoplastic Agents; pharmacology; Base Sequence; Chromosome Mapping; Chromosomes, Human, Pair 14; genetics; Cloning, Molecular; DNA, Complementary; chemistry; genetics; Drug Resistance, Multiple; genetics; Gene Expression Regulation, Leukemic; drug effects; HL-60 Cells; Humans; Leukemia; genetics; metabolism; pathology; Molecular Sequence Data; Multidrug Resistance-Associated Proteins; genetics; Oligonucleotide Array Sequence Analysis; Reverse Transcriptase Polymerase Chain Reaction; Sequence Analysis, DNA
- From: Chinese Journal of Medical Genetics 2005;22(2):158-163
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo find the novel gene related to the multi-drug resistance in leukemia and explore the molecular mechanism of multi-drug resistance.
METHODSThe 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.
RESULTSThe 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.
CONCLUSIONHV126, 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.