| Popis: |
Introduction: 2-dimensional (2D) cell culture is commonly used for the evaluation of anticancer drugs, which is incapable of simulating the three dimensions (3D) microenvironment of the original tumors, therefore, a new pre-clinical platform for drug screening is urgently needed. Material and Methods MDA-MB231 cells were grown either as 2D- monolayers or 3-dimensional (3D) spheroids and treated with 5-FU or doxorubicin. Cytotoxicity assays were performed using trypan blue exclusion dye for 2D- monolayers and 3D spheroids. We have evaluated the efficacy of 5-fluorouracil (5-FU) and doxorubicin at different concentrations on the growth, size of the spheroids, induction of cytotoxicity and cell death over 10 days. Shapiro-Wilk test was used for normal distribution and two-way ANOVA for multiple group comparisons. Results 2D- and 3D cultures responded with significant sensitivity toward the cytotoxic effect of 5-FU and doxorubicin. However, 3D spheroids were less sensitive to either of these drugs. The 3D spheroids exhibited significant variations in morphometric parameters in response to either 5-FU or doxorubicin treatment. The efficacy of doxorubicin was 1000-fold more effective than 5-FU over 10 days growth period of tumor spheroids. When compared to control spheroids both 5-FU and doxorubicin showed the presence of dead cells in the core of the spheroids as measured by live/dead assay. Discussion and Conclusion: Results indicated that 3D spheroid culture recapitulates in vivo tumor microenvironment and reflects the concentration of chemotherapeutic drugs required for effective treatment in cancer patients. Spheroid analysis using AnaSP/ReViSP could be further upgraded to include fluorescent stains within the workflow. |
