Related Experiment Videos
Test for chemotherapeutic sensitivity of cerebral gliomas: use of colorimetric MTT assay
J P Jordan1, C M Hand, R S Markowitz
1Department of Neurosurgery, Hahnemann University, Philadelphia, PA.
Abstract:
This study was undertaken to evaluate the colorimetric MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide] assay as a means of testing the sensitivity of gliomas to chemotherapeutic agents in vitro. Eight human glioma established cell lines were plated in 96-well tissue culture plates and incubated for 4 days with 10 different anti-cancer agents; 5 different concentrations of each drug were tested. The MTT dye was then added to the wells, and the resulting formazan precipitate was solubilized with dimethylsulfoxide (DMSO). The spectrophotometric absorbance (measured at 570 nm) of control and experimental wells was used to calculate the cytotoxicity index (CI). Values with a CI greater than 50% growth inhibition indicated cytotoxic efficacy (sensitivity to the chemotherapeutic drug). Six of the seven (85.7%) glioma cell lines were highly sensitive at varying concentrations to mitomycin C, cisplatin, and doxorubicin. Four of the seven (57.1%) cell lines demonstrated intermediate sensitivity to mitoxantrone and vinblastine. Five of the seven (71.4%) cell lines exhibited resistance to etoposide, bleomycin, cosmegen, and BCNU. One of the cell lines tested, U-138MG, failed to produce the MTT formazan precipitate, so that the sensitivity of this cell line to the panel chemotherapeutic drugs could not be determined. The variability of the results indicates the need for an in vitro screening method to evaluate the effectiveness of clinical and experimental chemotherapeutic agents. The MTT assay provides a rapid method of screening antineoplastic agents against gliomas for cytotoxicity.
Insights
The MTT assay effectively screens glioma cell lines for chemotherapy sensitivity in vitro. Most cell lines showed sensitivity to common agents like cisplatin, while others demonstrated resistance to different drugs.
Area of Science:
- Oncology
- Pharmacology
- Cell Biology
Background:
- Gliomas are aggressive brain tumors with limited treatment options.
- In vitro drug sensitivity testing is crucial for personalized glioma treatment.
- The MTT assay is a potential method for evaluating drug efficacy.
Purpose of the Study:
- To assess the colorimetric MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide] assay's utility for testing glioma sensitivity to chemotherapeutic agents.
- To determine the in vitro sensitivity of human glioma cell lines to various anti-cancer drugs.
Main Methods:
- Eight human glioma cell lines were cultured and exposed to 10 different chemotherapeutic agents at 5 concentrations each.
- The MTT assay was performed by adding MTT dye, solubilizing formazan precipitate with DMSO, and measuring spectrophotometric absorbance.
- Cytotoxicity index (CI) was calculated to determine drug sensitivity, with >50% growth inhibition indicating efficacy.
Main Results:
- 85.7% of glioma cell lines were highly sensitive to mitomycin C, cisplatin, and doxorubicin.
- 57.1% of cell lines showed intermediate sensitivity to mitoxantrone and vinblastine.
- 71.4% of cell lines were resistant to etoposide, bleomycin, cosmegen, and BCNU; one cell line's results were indeterminate due to assay failure.
Conclusions:
- The MTT assay is a rapid and effective method for screening antineoplastic agents against gliomas.
- Significant variability in drug sensitivity among glioma cell lines highlights the need for in vitro screening.
- This assay can aid in evaluating the effectiveness of both clinical and experimental chemotherapeutic agents for glioma treatment.