EGFP fluorescence as an indicator of cancer cells response to methotrexate Journal Article
| Authors: | Dabrowska, M.; Hendrikx, P. J.; Skierski, J.; Malinowska, M.; Bertino, J. R.; Rode, W. |
| Article Title: | EGFP fluorescence as an indicator of cancer cells response to methotrexate |
| Abstract: | Methotrexate action in viable cells was monitored by registering changes in EGFP (Enhanced Green Fluorescent Protein) fluorescence intensity. Treatment with 1 μM methotrexate for 48 h of human colorectal adenocarcinoma C85 cells, stably transfected to express EGFP, caused 5-fold increase in EGFP fluorescence assayed by flow cytometry with no distinct increase in EGFP protein level. This was correlated with morphological changes, including an increase of cell granularity and cell shape flattening, as well as cell cycle G1 phase arrest revealed by DNA content analysis. Methotrexate removal allowed the morphology of the cells in culture to revert in 10 days to normal. The cells that survived methotrexate exposure were propagated as C85r cell subline and displayed kinetics of methotrexate sensitivity parallel to that of the parental C85 line. As the increase in EGFP fluorescence could also be visualized by fluorescence microscopy, this reporter system may be employed to image methotrexate action in cancer cells in living models. © 2006 Elsevier B.V. All rights reserved. |
| Keywords: | controlled study; human cell; methotrexate; flow cytometry; adenocarcinoma; cell survival; cell structure; antimetabolites, antineoplastic; fluorescence; fluorescent dyes; cell line, tumor; colorectal carcinoma; colorectal neoplasms; dna; drug mechanism; cancer cell; green fluorescent proteins; microscopy, fluorescence; cell shape; folic acid antagonists; drug sensitivity; cell cycle g1 phase; enhanced green fluorescent protein; colonic carcinoma; imaging of drug response; cell granule; dna content |
| Journal Title: | European Journal of Pharmacology |
| Volume: | 555 |
| Issue: | 2-3 |
| ISSN: | 0014-2999 |
| Publisher: | Elsevier B.V. |
| Date Published: | 2007-01-26 |
| Start Page: | 93 |
| End Page: | 99 |
| Language: | English |
| DOI: | 10.1016/j.ejphar.2006.10.052 |
| PUBMED: | 17141212 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 3" - "Export Date: 17 November 2011" - "CODEN: EJPHA" - "Source: Scopus" |
