3D in Vitro Spheroid: A Preclinical Model for Drug Discovery Against Different Cancer

Nova Southeastern University, Davie, Florida, USA

Objective. This study was conducted to determine the effectiveness of the anti-cancer drug treatments using 3D (three-dimensional) cultures that closely mimic the tumor microenvironment (TME ) in the whole body. Background. The TME is one of the major determinants in regulating tumor progression and the metastatic process of cancers. The format of the cell growth can also determine the response of cancer cells to various drug treatments under the in vitro conditions. Methods. We conducted experiments with 3D culture to study the anti-cancer effects of SAHA, Nutlin-3, TMZ, and F16. The treatments were performed with lung (H1975 and H460), prostate (LNCaP-MST) and brain (U-87 MG) cancer cells that were growing 3D format as spheroids. Results. Western blot analysis of untreated cancer cells showed tenfold higher expression of cancer stem cell (CSC) markers (CD24, CD133) in the cells that were growing as spheroids in 3D cultures compared to the (2D) cultures. Also, the treatment of cells growing in 3D cultures as spheroids with SAHA (7.5 µM) lowered the expression of CSC markers in 7 days. Similar inhibition of CSCs was observed when the LNCaP-MST spheroids were treated with the MDM2 inhibitor Nutlin-3 (20 µM) for 7 days. Interestingly, the CSC markers showed a significant reduction in U-87 MG cells after F16 (a new drug patented by NSU) treatment also compared to TMZ. Conclusion. The 3D spheroid system may better predict the treatment outcomes of anticancer drugs. Grants. This research was funded by the Royal Dames of Cancer Research Inc., Florida.