Separating Response of Tumor and non-Tumor Cells to Drug
Antineoplastic Agents
/ pharmacology
Biomarkers, Tumor
/ antagonists & inhibitors
Cell Line, Tumor
Cell Survival
/ drug effects
DNA Mutational Analysis
/ methods
Dose-Response Relationship, Drug
Feasibility Studies
Humans
Melanoma
/ drug therapy
Mutation
Polymerase Chain Reaction
Proto-Oncogene Proteins B-raf
/ antagonists & inhibitors
Skin Neoplasms
/ drug therapy
Vemurafenib
/ pharmacology
BRAF
digital PCR
melanoma
mutation
mutation-quantification
Journal
Anticancer research
ISSN: 1791-7530
Titre abrégé: Anticancer Res
Pays: Greece
ID NLM: 8102988
Informations de publication
Date de publication:
Apr 2019
Apr 2019
Historique:
received:
19
02
2019
revised:
21
03
2019
accepted:
21
03
2019
entrez:
7
4
2019
pubmed:
7
4
2019
medline:
24
4
2019
Statut:
ppublish
Résumé
Conventional in vitro assays measure the effect of drugs on total cells, while separating the effect to those on tumor and non-tumor cells is important for assessing drug specificity. Our aim was to evaluate the feasibility of separating the efficacy of vemurafenib on tumor and non-tumor cells in a mixed culture. Melanoma A2058 cells and CCD18Co non-tumor cells were mixed and treated with vemurafenib. DNA was subjected to digital PCR to determine the ratio of the mutant 1799A to the wild-type 1799T alleles and viabilities of total cells were subsequently calculated as percentages of tumor and non-tumor cells. The set-up proportion of tumor cells correlated well with the calculated one. The calculated viability of tumor cells decreased with increasing doses of vemurafenib while that of the non-tumor cells remained rather constant. Variability of digital PCR data was high. Using the BRAF mutation 1799T>A to separate the response of tumor and non-tumor cells to a drug, such as vemurafenib, is feasible, supporting a foundation for a genetic in vitro tool for testing drug efficacy and specificity.
Sections du résumé
BACKGROUND/AIM
OBJECTIVE
Conventional in vitro assays measure the effect of drugs on total cells, while separating the effect to those on tumor and non-tumor cells is important for assessing drug specificity. Our aim was to evaluate the feasibility of separating the efficacy of vemurafenib on tumor and non-tumor cells in a mixed culture.
MATERIALS AND METHODS
METHODS
Melanoma A2058 cells and CCD18Co non-tumor cells were mixed and treated with vemurafenib. DNA was subjected to digital PCR to determine the ratio of the mutant 1799A to the wild-type 1799T alleles and viabilities of total cells were subsequently calculated as percentages of tumor and non-tumor cells.
RESULTS
RESULTS
The set-up proportion of tumor cells correlated well with the calculated one. The calculated viability of tumor cells decreased with increasing doses of vemurafenib while that of the non-tumor cells remained rather constant. Variability of digital PCR data was high.
CONCLUSION
CONCLUSIONS
Using the BRAF mutation 1799T>A to separate the response of tumor and non-tumor cells to a drug, such as vemurafenib, is feasible, supporting a foundation for a genetic in vitro tool for testing drug efficacy and specificity.
Identifiants
pubmed: 30952717
pii: 39/4/1777
doi: 10.21873/anticanres.13284
doi:
Substances chimiques
Antineoplastic Agents
0
Biomarkers, Tumor
0
Vemurafenib
207SMY3FQT
BRAF protein, human
EC 2.7.11.1
Proto-Oncogene Proteins B-raf
EC 2.7.11.1
Types de publication
Comparative Study
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1777-1783Informations de copyright
Copyright© 2019, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.