Reversal of multidrug resistance of K562/A02 cell line by mdr1 and GSTpi gene silence.
- Author:
Jing-wen GU
1
;
Tao ZHANG
;
Bo-bing CHEN
;
Yuan LU
;
Guo-wei LIN
Author Information
- Publication Type:Journal Article
- MeSH: ATP-Binding Cassette, Sub-Family B, Member 1; genetics; Drug Resistance, Multiple; genetics; Drug Resistance, Neoplasm; genetics; Genes, MDR; genetics; Genetic Vectors; Glutathione S-Transferase pi; genetics; Humans; K562 Cells; RNA Interference; RNA, Small Interfering; Transfection
- From: Chinese Journal of Hematology 2005;26(12):719-722
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effect of short hairpin RNA (shRNA) on mdr1 and GSTpi expression of human multidrug resistant leukemia cell line K562/A02.
METHODSshRNAs were synthesized according to the sequence targeting mdr1 and GSTpi coding region of 79-99nt and 308 approximately 327nt, and cloned into pSilencer 2.1-U6 neo vector. The cloned products, pSilence-mdr1 and pSilence-GSTpi, were transfected into K562/A02 cell line. Expression of mdr1 and GSTpi mRNA was assayed by real time PCR. 50% inhibition concentration (IC(50)) of doxorubicin (ADM) for K562/A02 cell line was determined by MTT method.
RESULTSAfter transfected with pSilence mdr1, the expression of mdr1 mRNA in K562/A02 cells was reduced by 71.5%, from (2.80 +/- 1.65) x 10(8) copy/microg RNA to (3.90 +/- 2.37) x 10(7) copy/microg RNA(P < 0.01). While the expression of GSTpi mRNA in pSilence-GSTpi transfected K562/A02 cells reduced by 39.8%, from (2.30 +/- 1.14) x 10(5) copy/microg RNA to (5.40 +/- 2.45) x 10(4) copy/microg RNA (P < 0.01). The resistance indexes after transfection were decreased to 8 and 10 respectively as compared to 23 of the mock transfection (P < 0.01).
CONCLUSIONThe shRNA could effectively reverse the multidrug resistance of K562/A02 cell line.
