I am trying to establish adriamycin-resistant MCF-7 from normal MCF-7 by sequential treatment of Adriamycin, but until now not succeeded in establishing it. Everytime results will fluctuate and most of the time, IC50 for both cell lines is the same. Even though, many reports to establish, I think its practically hard, please could you suggest to me the best way to establish r-MCF-7.
Hi, we have established a melanoma cell line resistant to vemurafenib (A375-VR8). My advice is to initially treat cells with an adriamycin concentration at least 10 times less than the IC50. Afterwards gradually increase the concentration, maintaining one MCF7 flask with the previous one (if cells will die because the second concentration is too high you don't need to restart from the beginning). The exposure time should be 1 to 2 weeks for every concentration, and you should reach an adriamycin concentration in the medium that is almost twice the original IC50. These cells will be always grown in presence od adriamycin.
Hope this could help!!
Thanks for your suggestions Luca and Phan. I am doing this work from past several months, actually IC50 of MCF-7 is 3 µM, so I started with 10 nM Adriamycin as per some literature, then by 20 nM, 40 nM, 100 nM, but the problem was when I treated first time with 10nM ADR, cells grow normally after 4 days when I replaced the medium with same concentration the growth was diminished and again by third time treatment almost stopped. I checked the IC50 of these cells, but resistivity is almost 2 folds or same as the parent cell. It is a part of my Ph.D. work, please could anybody suggest me where can I get these cell line or any good reference.
I perfectly understand your difficulty. You may contact Dr. Dihua Yu at MD Anderson Cancer Center. She is working on breast cancer drug resistance for many years. Her lab may have the cell line that you need. I am not so sure about it. The material transfer procedure may be very complicated but it may be worth trying.
Please send her a request. Here is her contact information
Dihua Yu, MD, PhD
Professor, Molecular & Cellular Oncology
The University of Texas MD Anderson Cancer Center
Email: [email protected]
Good luck Bomme :)
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