Detection of multidrug-resistance proteins with protein array chips.
- Author:
Bao-an CHEN
1
;
Juan DU
;
Chun-xiu ZHANG
;
Jian CHENG
;
Feng GAO
;
Zu-hong LU
Author Information
- Publication Type:Journal Article
- MeSH: ATP Binding Cassette Transporter, Sub-Family G, Member 2; ATP-Binding Cassette Transporters; analysis; ATP-Binding Cassette, Sub-Family B, Member 1; analysis; Drug Resistance, Multiple; genetics; Drug Resistance, Neoplasm; genetics; Humans; K562 Cells; Multidrug Resistance-Associated Proteins; analysis; Neoplasm Proteins; analysis; Protein Array Analysis
- From: Chinese Journal of Oncology 2005;27(9):528-530
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo evaluate the use of protein array chips in detection of multidrug-resistance proteins.
METHODSHuman erythroleukemic cell line K562 and its doxorubicin-resistant counterpart K562/A02 were used in the study. Monoclonal antibodies against P-glycoprotein (P-gP), multidrug resistance-associated protein (MRP1) and breast cancer resistance protein (BCRP) were immobilized onto agarose film-coated glass. The antibody-cell binding was assessed by capturing K562 and K562/A02 cells. The protein array was observed under a microscope and the image was captured with a CCD camera. The expression levels of the three proteins were also measured by flow cytometry (FCM).
RESULTSThe expression of P-gP and BCRP in K562 was very low. However, MRP1 expression was high. P-gP and MRP1 were highly expressed in K562/A02, while the expression of BCRP was low. FCM results showed that the expression rate of P-gP, MRP1 and BCRP in K562 cells was 5.98% +/- 2.19%, 95.80% +/- 3.98%, 1.03% +/- 0.45%, respectively, while that in K562/A02 cells was 92.67% +/- 1.80%, 97.18% +/- 1.02%, 3.98% +/- 0.37%, respectively. The results of protein array method are consistent with those of FCM (P > 0.05).
CONCLUSIONIt is feasible to develop a new protein array technique and to provide a novel method for multi-drug resistant cell detection, with a high throughput, high specificity, simple procedure and low cost.
