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Updated: Jun 24, 2025
Monitoring Cancer Cell Invasion and T-Cell Cytotoxicity in 3D Culture
Published on: June 23, 2020
Cytotoxic and molecular differences of anticancer agents on 2D and 3D cell culture
Mohammad Alwahsh1, Amani Al-Doridee2, Suhair Jasim2
1Faculty of Pharmacy, Al-Zaytoonah University of Jordan, Amman, 17138, Jordan. m.alwahsh@zuj.edu.jo.
Background:
Cancer and multidrug resistance are regarded as concerns related to poor health outcomes. It was found that the monolayer of 2D cancer cell cultures lacks many important features compared to Multicellular Tumor Spheroids (MCTS) or 3D cell cultures which instead have the ability to mimic more closely the in vivo tumor microenvironment. This study aimed to produce 3D cell cultures from different cancer cell lines and to examine the cytotoxic activity of anticancer medications on both 2D and 3D systems, as well as to detect alterations in the expression of certain genes levels.
Method:
3D cell culture was produced using 3D microtissue molds. The cytotoxic activities of colchicine, cisplatin, doxorubicin, and paclitaxel were tested on 2D and 3D cell culture systems obtained from different cell lines (A549, H1299, MCF-7, and DU-145). IC50 values were determined by MTT assay. In addition, gene expression levels of PIK3CA, AKT1, and PTEN were evaluated by qPCR.
Results:
Similar cytotoxic activities were observed on both 3D and 2D cell cultures, however, higher concentrations of anticancer medications were needed for the 3D system. For instance, paclitaxel showed an IC50 of 6.234 µM and of 13.87 µM on 2D and 3D H1299 cell cultures, respectively. Gene expression of PIK3CA in H1299 cells also showed a higher fold change in 3D cell culture compared to 2D system upon treatment with doxorubicin.
Conclusion:
When compared to 2D cell cultures, the behavior of cells in the 3D system showed to be more resistant to anticancer treatments. Due to their shape, growth pattern, hypoxic core features, interaction between cells, biomarkers synthesis, and resistance to treatment penetration, the MCTS have the advantage of better simulating the in vivo tumor conditions. As a result, it is reasonable to conclude that 3D cell cultures may be a more promising model than the traditional 2D system, offering a better understanding of the in vivo molecular changes in response to different potential treatments and multidrug resistance development.
Insights
Three-dimensional (3D) cell cultures, mimicking tumor microenvironments, show increased resistance to anticancer drugs compared to 2D cultures. This suggests 3D models offer better insights into in vivo cancer treatment and multidrug resistance.
Area of Science:
- Oncology
- Biotechnology
- Cell Biology
Background:
- Cancer and multidrug resistance pose significant health challenges.
- Two-dimensional (2D) cell cultures lack the complexity of the in vivo tumor microenvironment.
- Multicellular tumor spheroids (MCTS) and 3D cultures better replicate in vivo conditions.
Purpose of the Study:
- To generate 3D cell cultures from various cancer cell lines.
- To evaluate the cytotoxic effects of anticancer drugs on both 2D and 3D systems.
- To analyze gene expression alterations in response to treatments.
Main Methods:
- 3D cell cultures were created using microtissue molds.
- Cytotoxicity of colchicine, cisplatin, doxorubicin, and paclitaxel was assessed via MTT assay (IC50 determination).
- Gene expression of PIK3CA, AKT1, and PTEN was analyzed using qPCR.
Main Results:
- 3D cultures exhibited higher resistance to anticancer drugs, requiring increased concentrations for similar cytotoxic effects.
- Paclitaxel demonstrated significantly higher IC50 values in 3D H1299 cultures (13.87 µM) versus 2D (6.234 µM).
- Doxorubicin treatment led to a greater fold change in PIK3CA gene expression in 3D H1299 cultures compared to 2D.
Conclusions:
- Three-dimensional (3D) cell cultures demonstrate greater resistance to anticancer treatments than traditional 2D cultures.
- MCTS offer advantages in simulating in vivo tumor conditions due to their structure, growth patterns, and cellular interactions.
- 3D cell cultures represent a more promising model for understanding in vivo molecular changes and multidrug resistance development.
