|
[期刊论文][RESEARCH ARTICLE]
|
A Flow Cytometric Clonogenic Assay Reveals the Single‐Cell Potency of Doxorubicin
|
作 者:
|
Maass, Katie F.;Kulkarni, Chethana;Quadir, Mohiuddin A.;Hammond, Paula T.;Betts, Alison M.;Wittrup, Karl Dane;
|
|
出版年:2015
|
页 码:4409 - 4416
|
出版社:John Wiley & Sons, Ltd.
|
摘 要:
|
Standard cell proliferation assays use bulk media drug concentration to ascertain the potency of chemotherapeutic drugs; however, the relevant quantity is clearly the amount of drug actually taken up by the cell. To address this discrepancy, we have developed a flow cytometric clonogenic assay to correlate the amount of drug in a single cell with the cell's ability to proliferate using a cell tracing dye and doxorubicin, a naturally fluorescent chemotherapeutic drug. By varying doxorubicin concentration in the media, length of treatment time, and treatment with verapamil, an efflux pump inhibitor, we introduced 105–1010 doxorubicin molecules per cell; then used a dye-dilution assay to simultaneously assess the number of cell divisions. We find that a cell's ability to proliferate is a surprisingly conserved function of the number of intracellular doxorubicin molecules, resulting in single-cell IC50 values of 4–12 million intracellular doxorubicin molecules. The developed assay is a straightforward method for understanding a drug's single-cell potency and can be used for any fluorescent or fluorescently labeled drug, including nanoparticles or antibody–drug conjugates. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 104:4409–4416, 2015
|
关键字:
|
cancer chemotherapy;pharmacodynamics;drug effects;efflux pumps;cell lines
|
|
|
|
|
|
所属期刊
|
|
|
ISSN: 0022-3549
|
来自:John Wiley & Sons, Ltd.
|
|