The effect of Nano-curcumin on Cyclin D1 and DILA1 gene expression in breast cancer cells
The effect of Nano-curcumin on Cyclin D1 and DILA1 gene expression in breast cancer cells
Taraneh Givi,1Fatemeh Mohajerani,2Majid Sadeghizadeh,3,*
1. Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, P.O. Box: 14115-154, Tehran, Iran 2. Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, P.O. Box: 14115-154, Tehran, Iran 3. Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, P.O. Box: 14115-154, Tehran, Iran
Introduction: Breast cancer is the leading cause of cancer worldwide. It accounts for 22% of invasive cancers and 18% of all cancers in women. Breast cancer is a heterogeneous disease and there are differences in the response to various treatments, the risk of disease progression, and metastatic sites. About 20% of breast cancers have CCND1 gene amplification, and many irregularities in signaling pathways can lead to overexpression of CyclinD1 in about 50% of breast cancers, which are more of the ER + type. This project, study the expression of Cyclin D1, which is one of the most important oncogenes in breast cancer. There is also an LncRNA called DILA1, which interacts with Cyclin D1 and is overexpressed in breast cancer cells.DILA1 specifically binds to Ther286 of Cyclin D1 protein and inhibited its phosphorylation, leading to decreased ubiquitination and degradation of Cyclin D1. Recent investigations in cancer treatment have revealed curcumin anti-cancer properties through different pathways. Nanotechnology has been employed to overcome this barrier. Nano-formulated curcumin (Curcuden) has been shown to provide a significantly higher bioavailability for oral consumption.
Methods: MCF-7 cells were cultured in DMEM, supplemented with 10 % fetal bovine serum and 1% Pen-Strep. 2×105cells were planted per well in a 6-well plate in duplicate. After 24h cells were treated with nano-curcumin(17μM). Next, in 24h and 48h total RNAs were extracted with Trizol Reagent. The quality of RNA was measured by gel electrophoresis. cDNAs were synthesized using the manuscript’s protocol. Expression of Cyclin D1, DILA1, BAX, and BCL2 genes were measured by qRT PCR in treated and untreated MCF-7 cells.
Results: Results: To compare the expression level of Cyclin D1 and DILA1 in untreated, 24h and 48h nano-curcumin treated cells qPCR was done. Expression of Cyclin D1 and DILA1 was decreased at 24h and 48h after being treated with nano-curcumin compered to untreated cells. As expected the reduction of these genes was much higher in 48h than 24h after treatment. To study the effect of nano-curcumin on the apoptotic pathway, the expression level of apoptotic genes was determined by qRT PCR. The results confirmed that nano-curcumin is apoptosis inducer by downregulation of BCL2 and upregulation of BAX gene.
Conclusion: These results indicate that nano-curcumin was able to significantly reduce the DILA1expression leading to destabilizing of the Cyclin D1 protein. Therefore, reducing DILA1 expression induces apoptosis in breast cancer cells.According to these results, nano-curcumin induces apoptosis in breast cancer cells by downregulation of Bcl2 and upregulation of BAX gene.
Nanocurcumin has a relatively cytotoxic effect on MCF-7 breast cancer cells, suppressing the expression of Cyclin D1, a critical gene in the development and metastasis of breast cancer.