Journal of Mazandaran University of Medical Sciences (Jul 2024)
Evaluation of Synergic Effect of Nano-Micelle Curcumin and Epicatechin on Apoptosis and Proliferation of HT29 Colorectal Cancer Cell Line
Abstract
Background and purpose: Today, despite the abundance of chemical drugs, the use of medicinal plants is increasing; Curcumin (curcumin) is a yellow pigment obtained from the turmeric plant and is considered the effective ingredient of turmeric. This crystalline and crystallized product is widely used in Indian medicine. Recently, the beneficial effects of curcumin on stomach, colon, and mouth cancers have been proven in mice. Curcumin usually plays its role through pharmacological processes such as antioxidant, anti-inflammatory, antithrombotic, apoptotic, and hepatoprotective effects. Antioxidative, antiproliferative, and antiangiogenic properties of curcumin have been noticed in recent years. However, the low solubility of curcumin and its severe degradation have made it impossible to introduce it in clinical use. In recent years, studies and reviews have been conducted to prepare and design polymeric micelles (PMMCs) on a nanoscale to improve the cellular transport of curcumin. Green tea flavonoids, including epicatechin, have recently received attention. In in vitro research, these compounds have been proposed as antioxidants by inhibiting lipid peroxidation, trapping free radicals, and chelating metal ions, and this property is mentioned due to their unique structure (therefore, the purpose of this study is to determine the effect The synergism of curcumin and epicatechin has been carried out in the investigation of apoptosis and cell proliferation of HT29 cell lines and apoptotic factors. Materials and methods: This study is experimental. After obtaining HT29 cells from the Pasteur Institute cell bank and completing cell passage and proliferation, the cells were distributed in triplicate wells across 7 test groups and 2 control groups (positive and negative). In test groups 1, 2, and 3, three concentrations of curcumin nanomicelles (10, 20, 50 µg/ml) were incubated in triplicate. In groups 4, 5, and 6, three concentrations of epicatechin (10, 20, 50 µg/ml) were incubated for 24 hours. Test group 7 consisted of HT29 cells treated with the IC50 concentration of nanomicelle curcumin combined with the IC50 concentration of epicatechin. Group 8, the positive control, consisted of cells treated with 5FU (100 µg/ml), while group 9, the negative control, consisted of cells exposed only to the cell culture medium. After the 24-hour incubation period, an MTT assay was performed to assess cell proliferation, and Annexin flow cytometry was used to determine the level of apoptosis. Results: In this study, epicatechin and curcumin have been able to decrease cell viability of colon cancer cells (HT29) and increase the percent of apoptotic cells P<0.05, which was in a dose-dependent manner, also the combination of these two drugs caused a synergistic effect in high dose P<0.05. Epicatechin in concentrations of 20, 10 μg/mL, 50, and 100 has a significant difference compared to the negative control group with P<0.05, which has caused a decrease in cell viability in colon cancer cells with increasing dose. Curcumin is the group Nanomicelle with epicatechin at IC50 concentration compared to the positive control group of 5-FU in HT-29 cell line has greatly reduced cell viability of HT-29 cell line showed that curcumin and epicatechin nano micelles had a synergistic effect. The results of cell apoptosis have shown that curcumin and epicatechin nano micelle groups at IC50 concentrations had a synergistic effect. Conclusion: epicatechin and curcumin have been able to increase the death rate of colon cancer cells (HT29) by increasing apoptosis, which indicates the anti-cancer effects of these compounds. So it can suggest these compounds in colon cancer therapeutics.
