HA117 gene increased the multidrug resistance of K562 cells in vitro: an investigation to the function of a novel gene related to drug resistance.
OBJECTIVE
A novel multi-drug resistance gene named as HA117 has been screened and cloned in multidrug resisitant leukemia cell lines in our previous research, but its function is still unknown. In this study, HA117 gene was investigated whether it could increase the drug resistance in chronic myelogenous myeloid leukemia cell line K562.


METHODS
HA117 was cloned and adenovirus vectors were constructed with the HA117 gene (Adeasy-HA117). K562 cells were infected by Ad-HA117 to get the K562/Ad-HA117 cells with HA117 gene expression. The infection efficiency and the multiplicity of infection (MOI) were detected by fluorescence and flow cytometry. The expression of HA117 gene was detected by RT-PCR. The drug sensitivities of K562/Ad-HA117 cells were detected by Methyl Thiazolyl Tetrazolium (MTT) assay.


RESULTS
Recombinant adenovirus vectors were constructed and a MOI of 100 is most suitable to infect K562 cells. The infected K562 cells demonstrated in vitro production of HA117 mRNA as measured by reverse-transcriptase polymerase chain reaction. There were no significant changes in K562/Ad-HA117 cells growth, while the drug sensitivities of K562/Ad-HA117 cells to Vincristine, Adriamycin, Etoposide, Daunorubicin, Mitomycin and Cyclophosphamide decreased 4.44, 7.18, 3.01, 9.53, 3.48 and 3.61 times than that of uninfected K562 cells, respectively (P < 0.05).


CONCLUSIONS
Expression of the novel gene HA117 could significantly increased the multi-drug resistance of K562 cells, which indicated that HA117 is a functionally relevant multidrug resistance gene.
Guo
Yuxia
Y
The Affiliated Children's Hospital, Chongqing Medical University, PR China. guoyuxia9955@yahoo.com.cn
Zheng
Gaihuan
G
Jin
Xianqing
X
Xu
Youhua
Y
Luo
Qing
Q
Liu
Xiaomei
X
Zhao
Zhenzhen
Z
Chen
Yong
Y
D000256:Adenoviridae / Q000235:genetics; D000970:Antineoplastic Agents / Q000494:pharmacology; D018432:Drug Resistance, Multiple / Q000187:drug effects* / Q000235:genetics*; D019008:Drug Resistance, Neoplasm / Q000187:drug effects* / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects* / Q000235:genetics*; D006801:Humans; D020014:K562 Cells; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013379:Substrate Specificity
D000970:Antineoplastic Agents