The Selective Cytotoxicity of Quercus Brantii Lindl. Galls on A375 and SK-MEL-3 Human Malignant Melanoma Cell Lines.

DOI:10.31557/APJCP.2023.24.7.2383 Anti-Cancer Effect of Quercus brantii Lindl. galls RESEARCH ARTICLE Editorial Process: Submission:02/19/2023 Acceptance:07/07/2023 The Selective Cytotoxicity of Quercus Brantii Lindl. Galls on A375 and SK-MEL-3 Human Malignant Melanoma Cell Lines Bahareh Sadat Yousefsani1, Kamyar Mohajer2, Ali Qobadi1, Elahe Aghazadeh3, Kobra Shirani3*, Jalal Pourahmad2* Abstract This study aimed to find out the mechanism of cytotoxic effects of galls of Quercus Brantii on A375 and SKMEL-3 melanoma and AGO-1522 normal human fibroblast cell lines for the first time. Therefore, cell viability and cytotoxic activities were evaluated. Furthermore, ROS formation, lipid peroxidation, and release of cytochrome-c were also assessed. The results revealed that the extract of these galls at a concentration of 0.05 mg/ml significantly (P<0.001) increased cytotoxicity, ROS formation, TBARS formation, and cytochrome-c release in A375 and SK-MEL-3 melanoma cell lines compared to AGO-1522 normal human fibroblast. These results demonstrated that these galls can be considered a promising candidate which acts in synergy with anticancer agents used in the clinical treatment of human malignant melanoma. Keywords: Melanoma- Quercus- cytotoxicity- skin cancer Asian Pac J Cancer Prev, 24 (7), 2383-2388 Introduction Malignant melanoma is a highly malignant and invasive skin tumor, which is the most fatal skin cancer (Habif, 2015). In recent decades, the worldwide incidence of that cancer has increased rapidly. Researches indicate that due to the aging of the population, melanoma prevalence is doubled every 10 to 20 years (Garbe and Leiter, 2009; Liu et al., 2016). Environmental factors including exposure to sunshine (especially type B ultraviolet radiation), occupational and nutritional factors, as well as multiple and abnormal nevi and immunosuppression are melanoma risk factors (Jiang et al., 2015; Noonan et al., 2012). Besides, genetic and epigenetic factors play an important role in this disease (Caini et al., 2017; Chen et al., 2015; Gandini et al., 2005). There are many efforts for reaching effective treatment for melanoma. Routine therapies include surgery, chemotherapy, hormone therapy, radiation therapy, gene therapy, and immunotherapy. Surgery is the first-line treatment for local lesions, especially in the early stage. However, in advanced cases, due to the metastatic nature of this cancer, response to treatment is weak. Melanoma is resistant to common medications such as single-drug chemotherapy regimens and radiotherapy, therefore, suggesting novel, effective, and low-risk therapies (Guy Jr et al., 2015). The usage of medicinal plants in cancer therapy regimens has always been an interesting topic for research (Cragg and Newman, 2005). However, the herbal compounds and antioxidants in cancer therapy is still an important issue. But research has shown that nutritional advice provided to patients at the right time and under the supervision of a specialist can be somewhat helpful to treatment during the course of the disease and reduce drug side effects (Richardson, 2001; Tonkaboni et al., 2021). Researches have demonstrated the synergistic effects of natural products with various chemotherapeutic medications used to treat cancer (Guney Eskiler et al., 2019; Yurdacan et al., 2019). In a study by Abu Hazafa and his colleagues, the use of herbal compounds and especially polyphenols could be helpful in the treatment of various cancers through molecular mechanisms (Hazafa et al., 2020). Previous studies reported the effectiveness of natural products in the treatment of malignant melanoma (Gladfelter et al., 2017; Iranzadasl et al., 2021). Quercus Brantii Lindl. (Oak), belongs to the family of Fagaceae. They are medium or short trees in size. There are galls (locally called ‘mazoo’) that arise on its younger branches which are caused by the gall wasp Adleria gallae-tinctoria offense (Samuelson and Bohlin, 1992). These trees are widespread in provinces like Lorestan and Kordestan in Iran. Tannins, gallic acid, syringic acid, ellagic acid, β-sitosterol, amentoflavone, ether,and isocryptomerin are some of the main gall compounds. Department of Traditional Pharmacy, School of Persian Medicine, Iran University of Medical Sciences, Tehran, Iran. Department of Toxicology and Pharmacology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran. 3Department of Toxicology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. *For Correspondence: , 1 2 Asian Pacific Journal of Cancer Prevention, Vol 24 2383 Bahare Yosefsani et al (Kaur et al., 2004). Quercus Brantii galls are used for decades in Persian medicine for treating different types of inflammation (Aghili, 2009). Previous researches have been reported that these galls have many pharmacological effects like: astringent, anti-pyretic, anti-parkinsonism (Kaur et al., 2004), local anesthetic, central nervous system depressant, analgesic (Dar et al., 1976; Kaur et al., 2004), anti-diabetic (Hwang et al., 2000; Kaur et al., 2004), anti-bacterial (Rodríguez and Rull, 1980; Sawangjaroen et al., 2004), anti-oxidant (Hamid et al., 2005; Kaur et al., 2008) and anti-inflammatory (Kaur et al., 2004). In this mechanistic cellular study, we wanted to demonstrate whether the hydroalcoholic extract of galls could be helpful in the treatment of human malignant melanoma. Materials and Methods Chemicals 2,4,6-Trinitrobenzene sulfonic acid (TNBS) and rhodanine from Sigma-Aldrich Chemie (GmbH, Munich, Germany), thiobarbituric acid (TBA), trichloroacetic acid (TCA), n-butanol, hexadecyl trimethyl ammonium bromide (HETAB), 2,4,6-tri (2-pyridyl)-s-triazine (TPTZ), diphenyl-2-picryl hydrazyl (DPPH), methanol, hydrochloric acid (HCl), malondialdehyde (MDA), ethylenediamine tetra-acetic acid (EDTA), O-di anisidine hydrochloride, hydrogen peroxide, acetic acid, sodium acetate, Coomassie reagent, bovine serum albumin (BSA), ferric chloride (FeCl3 · 6 H2O),sodium sulfate (Na2SO4), sulfuric acid (H2SO4),phosphoric acid (H3PO4), potassium dihydrogen phosphate (KH2PO4), potassium hydrogen diphosphate (K2HPO4), peroxide hydrogen (H2O2) and sodium carbonate (Na2CO3) from Merck (Germany) and galls of Quercus Brantii were purchased from Iran. Preparation of Plant Sample The galls of Quercus Brantii were purchased from the market of medicinal plants in Tehran, Iran, in December 2020. They looked fresh and ground well (Fig. 1). The document of the sample was preserved in the Herbarium of the Department of Pharmacognosy, Faculty of Pharmacy, Tehran University of Medical Science, Tehran, Iran (No. 6730-TEH). The dried galls of Quercus Brantii were powdered. The ethanolic extract was prepared by mixing the 30 g of mentioned powder with ethanol/water (70:30) mixture (21.8 g dry weight corresponding to 73% w/w) (Alizade Naini et al., 2021). Cell culture All procedures were conducted according to the ethical sta (...truncated)