Functional Chemoresistance Characterization: A Method to Evaluate Drug Resistance in Cancer Cells Using Clonogenic Assay
Published: April 30, 2023
Abstract
Source: Mohr, L. et al. Generation of Prostate Cancer Cell Models of Resistance to the Anti-mitotic Agent Docetaxel. J. Vis. Exp. (2017)
This video describes a method for evaluating chemotherapeutic drug resistance in prostate cancer cells using colony formation assay. This assay can be used as a functional tool to identify novel molecular and cellular mechanisms of chemoresistance in cancer cells.
Protocol
1. Functional Characterization of Acquired Docetaxel Resistance by Colony Formation Assay NOTE: In this protocol, chemoresistance has been evaluated using colony formation assays. Alternative methods used to evaluate cell viability (i.e., MTS assays) can be used based on investigators' preferences. Plate 2,000 cells (DU145 or 22Rv1, both parental and Docetaxel-resistant) per well in 6-well plates, using 2 mL of media per w…
Representative Results
Figure 1: Functional and phenotypic characterization of the Docetaxel-resistant cell models. (A) Representative colony formation assays of parental and Docetaxel-resistant cells treated with the indicated Docetaxel concentrations for 72 h. Percentage of colonies for every treatment concentration is represented in the included graph. The experiments are triplicates and data r…
Divulgations
The authors have nothing to disclose.
Materials
| DU145 cells | ATCC | HTB-81 | |
| 22Rv1 cells | ATCC | CRL-2505 | |
| Docetaxel | Selleck Chemicals | S1148 | in DMSO (10mM) |
| Tissue culture flask 150 sq cm | Falcon | 355001 | |
| 6-well tissue culture plates | Falcon | 353046 | |
| RPMI 1640 Medium | Gibco | 11875093 | Supplemented with 10% FBS and 1% Penicilin/Streptomycin |
| Foundation B Fetal Bovine Serum | Gemini | 900208 | |
| 1X Phosphate-Buffered Saline (PBS) | Corning | 21-040-CM | |
| 0.05%Trypsin-EDTA | Gibco | 25300-062 | |
| Penicillin-Streptomycin | Gibco | 15140122 | |
| Crystal Violet | Sigma-Aldrich | C0775 | |
| 10% Formalin Solution | Sigma-Aldrich | HT501128 |
Citer Cet Article
Functional Chemoresistance Characterization: A Method to Evaluate Drug Resistance in Cancer Cells Using Clonogenic Assay. J. Vis. Exp. (Pending Publication), e20400, doi: (2023).