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Published on: June 13, 2020
Evaluation of Combined Treatment With Atorvastatin and SREBP2 Inhibitors Against Colorectal Cancer Cells Under
Kyota Ishii1, Yuno Tauchi2, Tomohiro Yano2,3
1Laboratory of Molecular Bromacology, Graduate School of Health and Sports Sciences, Toyo University, Tokyo, Japan; s4h202300016@toyo.jp.
Background/Aim:
Statins, inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, have been reported to exert anti-cancer effects. Recently, sterol regulatory element-binding protein 2 (SREBP2) has been shown to be involved in statin resistance in cancer cells. Inhibiting SREBP2 enhances the anti-cancer effects of statins in several cancer cell lines. Three-dimensional (3D) cultured cells are known to exhibit different drug sensitivities compared to two-dimensional (2D) cultured cells. In this study, we evaluated the cytotoxicity induced by combined treatment with atorvastatin and SREBP2 inhibitors in 2D and 3D cultured colorectal cancer (CRC) cells.
Materials And Methods:
25-Hydroxycholesterol and δ-tocotrienol were used as SREBP2 inhibitors. The viability of 2D and 3D cultured cells was measured using the MTT assay and the CellTiter-Glo® 3D Cell Viability Assay. Gene expression levels were analyzed by qRT-PCR. Protein levels were determined by western blotting.
Results:
3D cultured cells exhibited lower statin sensitivity compared to 2D cultured cells. The expression levels of SREBP2 and its target genes differed between 2D and 3D cultured cells. In 2D cultured cells, SREBP2 inhibitors blocked atorvastatin-induced SREBP2 activation and enhanced the cytotoxicity of atorvastatin. However, in 3D cultured cells, SREBP2 inhibitors failed to enhance atorvastatin-induced cytotoxicity, although they successfully suppressed atorvastatin-induced SREBP2 activation.
Conclusion:
The efficacy of a combined statin and SREBP2 inhibitors strategy differs substantially between 2D and 3D cultured CRC cells. 3D cultured cells may possess an SREBP2-independent mechanism of statin resistance that differs from that of 2D cultured cells.
Insights
Combining statins with SREBP2 inhibitors shows different effects on colorectal cancer cells grown in 2D versus 3D cultures. Three-dimensional cultures exhibit statin resistance potentially independent of SREBP2, impacting treatment strategies.
Area of Science:
- Oncology
- Pharmacology
- Cell Biology
Background:
- Statins, 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors, possess anti-cancer properties.
- Sterol regulatory element-binding protein 2 (SREBP2) is implicated in cancer cell statin resistance.
- Inhibiting SREBP2 can potentiate statin efficacy in various cancer cell lines.
Purpose of the Study:
- To investigate the cytotoxicity of combined atorvastatin and SREBP2 inhibitor treatment.
- To compare treatment effects in 2D versus 3D cultured colorectal cancer (CRC) cells.
- To explore the role of SREBP2 in statin resistance in different culture models.
Main Methods:
- Utilized 25-hydroxycholesterol and δ-tocotrienol as SREBP2 inhibitors.
- Assessed cell viability using MTT and CellTiter-Glo assays for 2D and 3D cultures.
- Analyzed gene and protein expression of SREBP2 and its targets via qRT-PCR and Western blotting.
Main Results:
- 3D cultured CRC cells demonstrated reduced sensitivity to statins compared to 2D cultures.
- SREBP2 inhibitors enhanced atorvastatin cytotoxicity in 2D cultures by blocking SREBP2 activation.
- In 3D cultures, SREBP2 inhibitors did not improve atorvastatin efficacy despite suppressing SREBP2 activation.
Conclusions:
- The combined statin and SREBP2 inhibitor strategy's effectiveness varies significantly between 2D and 3D CRC cell models.
- 3D cultured cells may employ SREBP2-independent mechanisms for statin resistance.
- Findings suggest distinct resistance pathways in 3D versus 2D cancer models warranting further investigation.
