Comparison of anti-cancer effects of Lentinan and Docetaxel nanofibers on the expression of cancer-inducing genes
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Hanieh Ataollahi,1 Mohaddeseh Larypoor,2,*
1. Faculty of Biological Science, Department of Biotechnology, Islamic Azad University, North Tehran Branch, Tehran, Iran.
2. Faculty of Biological Science, Department of Biotechnology, Islamic Azad University, North Tehran Branch, Tehran, Iran.
- Introduction: Lentinula edodes, also known as Shiitake, is a native basidiomycete that is both edible and medicinal. This Mushroom plays a considerable role in cancer therapy as an adjuvant and enhancing the quality of life of patients due to its bioactive compounds and polysaccharides with anti-cancer and anti-tumor properties such as Lentinan in China and Japan. Lentinan rigid polysaccharides can participate in the fabrication of various types of nanocomposites. The use of compounds from this medicinal mushroom in cancer treatment will be useful because they will suppress the expression of genes that are effective in causing cancer along with current medicines. HER3 is a member of the epidermal growth factor receptor (EGFR) family, which is overexpressed in 20-30% of invasive breast cancers. This gene forms a heterodimer with other HER family members. Breast cancer is highly prevalent among women all over the world. Therefore, the use of natural and native compounds with high effectiveness without side effects along with commonly used antineoplastic drugs (which have various side effects, are often costly, and out of reach of patients) would have synergistic effects in reducing the invasion of cancer cells.
- Methods: First, the optimal conditions for solid and liquid culture media, acidity and temperature for the growth of Lentinula edodes (TMU340) mycelium were determined. Also, by using the lyophilization process, the ratio of wet to dry weight of mycelium and fruiting bodies of this mushroom was determined. Isolation and extraction of Lentinan polysaccharide were performed using 60 °C hot water method, deproteinization by Sevage method, extraction, and precipitation with absolute ethanol from mycelium and fruiting body; By using the DEAE Sephadex A-25 ion-exchange chromatography, polysaccharide was purified and the concentration was obtained by phenol-sulfuric acid test. Then electrospinning was used to spin nanofibers, and SEM and FTIR were used to study the morphology and composition of these nanofibers. MCF-7 cancer cells were treated with different concentrations of treatment groups, including PVA /Lentinan /Docetaxel, PVA/Docetaxel and PVA/Lentinan nanofibers with positive control of Docetaxel, Lentinan, and PVA. MTT Assay was used to calculate the cell viability percentage and expression of the HER3 gene was determined using Real-Time PCR compared to untreated controls.
- Results: The optimal liquid and solid culture media were determined as PDA and PDB media, respectively. Also, the optimum temperature and pH for the growth of mycelium of this mushroom was determined as 25 °C and pH 5.5 during 14 days of growth. Wet to dry weight ratio in the mycelium and the fruiting body was calculated as a 10:1 ratio. The results of the phenol-sulfuric acid test determined a higher concentration of Lentinan in mycelial extraction than the fruiting body, equal to 0.243 mg/ml. Optimal electrospinning conditions were determined for 1 kV, 15 cm distance, and 1 ml/h flow rate. The cell viability percentage of MCF-7 cells were reported as 28.5 percent for 72 hours after treatment with triple PVA/Lentinan/Docetaxel nanofibers. The expression level of the HER3 gene was 0.52 compared to the control group at the most effective concentration for 72 hours; Also, the expression level of this gene by the treatment of free Lentinan was 0.9 and free Docetaxel was 0.62, which indicates a synergistic effect and suitable for drug supplementation.
- Conclusion: Lentinula edodes is an inexpensive, affordable, easy to grow, and native mushroom. The compounds and bioactive polysaccharides of this mushroom have given special medicinal value to it. The reducing effect of polysaccharides of this mushroom on the expression of the HER3 gene can be effective in preventing the progression of cancer by controlling the expression of genes involved in cancer with conventional chemotherapy drugs. Triple PVA/Lentinan/Docetaxel nanofibers have a suitable synergistic effect over time, which has a favorable anti-cancer effect by reducing the expression of the HER3 gene and is a suitable drug delivery system.
- Keywords: Lentinan, Electrospinning, Breast cancer, Shittake, Docetaxel
