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…

Divulgaciones

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

Play Video

Citar este artículo
Functional Chemoresistance Characterization: A Method to Evaluate Drug Resistance in Cancer Cells Using Clonogenic Assay. J. Vis. Exp. (Pending Publication), e20400, doi: (2023).

View Video