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Updated: Jun 19, 2026
Three-Dimensional (3D) Tumor Spheroid Invasion Assay
Published on: May 1, 2015
3D imaging of the response to CDC25 inhibition in multicellular spheroids
Céline Frongia1, Corinne Lorenzo, Fréderic Gianni
1Centre National de la Recherche Scientifique, LBCMCP UMR5088, Toulouse, France.
Abstract:
Multicellular tumor spheroids closely mimic the 3D organization of avascular microregions within tumors and thereby represent a valuable model for the evaluation of anticancer drugs. In this study, we performed a 3D analysis of the response to the CDC25 phosphatase inhibitor IRC-083864 in HCT116 spheroids. Continuous exposure to IRC-083864 strongly inhibits the growth of spheroids and is shown to correlate with a decrease in Ki-67 positive cells. The cytotoxicity induced by IRC-083864 was examined by two-photon laser microscopy imaging and 3D reconstruction. Visualization in 3D allowed us to demonstrate that IRC-083864 treatment results in the inhibition of mitosis and induces cell death specifically localized in the outer proliferative cell layers of the spheroid structure. These results emphasize the importance of 3D models and of in toto analysis for the evaluation of anticancer drugs cytotoxicity.
Insights
This study shows that the CDC25 phosphatase inhibitor IRC-083864 effectively inhibits HCT116 spheroid growth and induces cell death in the outer layers. Three-dimensional analysis highlights its potential as an anticancer drug.
Area of Science:
- Oncology
- Pharmacology
- Biotechnology
Background:
- Multicellular tumor spheroids accurately model avascular tumor regions.
- Spheroids are valuable for anticancer drug evaluation.
- Three-dimensional (3D) analysis offers deeper insights into drug responses.
Purpose of the Study:
- To investigate the 3D response of HCT116 spheroids to the CDC25 phosphatase inhibitor IRC-083864.
- To evaluate the cytotoxicity and growth inhibition effects of IRC-083864.
- To demonstrate the utility of 3D models for drug assessment.
Main Methods:
- Culturing HCT116 cells into multicellular tumor spheroids.
- Treating spheroids with the CDC25 phosphatase inhibitor IRC-083864.
- Utilizing two-photon laser microscopy and 3D reconstruction for analysis.
- Assessing cell proliferation (Ki-67) and cell death.
Main Results:
- Continuous IRC-083864 exposure significantly inhibited spheroid growth.
- A decrease in Ki-67 positive cells correlated with IRC-083864 treatment.
- 3D visualization revealed mitosis inhibition and cell death localized to outer spheroid layers.
- The drug's cytotoxicity was confirmed within the 3D spheroid structure.
Conclusions:
- IRC-083864 demonstrates potent anticancer activity in a 3D spheroid model.
- Three-dimensional analysis is crucial for understanding drug mechanisms and cytotoxicity.
- This study underscores the importance of 3D models for preclinical anticancer drug evaluation.
