Functional expression and characterization in Xenopus laevis oocytes of the ABCG2 transporter derived from A549 human lung adenocarcinoma cells
| Autor: | Hee Yi, Sang Min Jeong, Seung-Yeol Nah, Dong-Goo Lee, Hee-Jung Cho, Soo-Min Cho, Ho-Chul Shin, Byung-Hwan Lee, Sung-Hee Hwang, Kyul Jo, Jin-A Park |
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| Rok vydání: | 2011 |
| Předmět: |
Cancer Research
Antimetabolites Antineoplastic animal structures DNA Complementary Lung Neoplasms Abcg2 Transcription Genetic Cell Molecular Sequence Data Xenopus ATP-binding cassette transporter Adenocarcinoma of Lung Adenocarcinoma Xenopus laevis Side population Cell Line Tumor medicine ATP Binding Cassette Transporter Subfamily G Member 2 Animals Humans Amino Acid Sequence A549 cell biology Base Sequence Cell Membrane General Medicine Oocyte biology.organism_classification Molecular biology Neoplasm Proteins Protein Transport medicine.anatomical_structure Methotrexate Oncology Cell culture embryonic structures biology.protein ATP-Binding Cassette Transporters Female sense organs Sequence Alignment Plasmids |
| Zdroj: | Oncology reports. 27(2) |
| ISSN: | 1791-2431 |
| Popis: | We cloned the ATP-binding cassette sub-family G member 2 (ABCG2) transporter, the most recently identified among several major human multidrug-resistance pumps, from A549 human lung adenocarcinoma cells in order to characterize its function and substrate specificity. In a previous report, we confirmed that a stem cell-like side population of A549 cells highly expressed the ABCG2 gene and had a unique ability to resist the anticancer drug methotrexate (MTX). In this study, ABCG2 cDNA was cloned by RT-PCR and converted into cRNA by an in vitro transcription system for expression in Xenopus laevis (X. laevis) oocytes. The transcribed cRNA of the ABCG2 gene was injected into the oocytes under the absence of cofactors or heterologous partner proteins or some lipids from the media. A high expression of ABCG2 was observed on the oocyte surface by immunofluorescence and confocal laser microscopy. We tested the functional effect of ABCG2 expression on drug efflux by directly injecting MTX into X. laevis oocytes. The drug concentration within the oocytes was quantified with LC-MS/MS; the analysis showed that the accumulation of MTX was significantly decreased in the X. laevis oocytes expressing ABCG2 compared with the control oocytes not expressing ABCG2. These findings show that the ABCG2 protein has an important role in the efflux of MTX through the cell membrane of X. laevis oocytes. Therefore, it might be that ABCG2, abundantly expressed in the stem cell population of A549 cells, can modulate resistance to MTX in lung cancer therapy. |
| Databáze: | OpenAIRE |
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