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