Department of Biochemistry, Faculty of Pharmacy, University of Santo Tomas, Manila, Philippines.
Research Center for Natural and Applied Sciences, University of Santo Tomas, Manila Philippines.
Asian Pac J Cancer Prev. 2020 Aug 1;21(8):2423-2430. doi: 10.31557/APJCP.2020.21.8.2423.
Breast cancer is a multifactorial disease that affects women worldwide. Its progression is likely to be executed by oxidative stress wherein elevated levels of reactive oxygen and nitrogen species drive several breast cancer pathologies. Spider venom contains various pharmacological peptides which exhibit selective activity to abnormal expression of ion channels on cancer cell surface which can confer potent anti-cancer activities against this disease.
Venom was extracted from a Philippine tarantula by electrostimulation and fractionated by reverse phase-high performance liquid chromatography (RP-HPLC). Venom fractions were collected and used for in vitro analyses such as cellular toxicity, morphological assessment, and oxidative stress levels.
The fractionation of crude spider venom generated several peaks which were predominantly detected spectrophotometrically and colorimetrically as peptides. Treatment of MCF-7 cell line of selected spider venom peptides induced production of several endogenous radicals such as hydroxyl radicals (•OH), nitric oxide radicals (•NO), superoxide anion radicals (•O2-) and lipid peroxides via malondialdehyde (MDA) reaction, which is comparable with the scavenging effects afforded by 400 µg/mL vitamin E and L-cysteine (p<0.05). Concomitantly, the free radicals produced decrease the mitochondrial membrane potential and metabolic activity as detected by rhodamine 123 and tetrazolium dye respectively (p>0.05). This is manifested by cytotoxicity in MCF-7 cells as seen by increase in membrane blebbing, cellular detachment, caspase activity and nuclear fragmentation.
These data suggest that the Philippine tarantula venom contains peptide constituents exhibiting pro-oxidative and nitrosative-dependent cytotoxic activities against MCF-7 cells and can indicate mechanistic insights to further explore its potential application as prooxidants in cancer therapy.
.
乳腺癌是一种影响全球女性的多因素疾病。其进展可能是由氧化应激引起的，其中活性氧和氮物种水平升高会导致几种乳腺癌病理发生。蜘蛛毒液含有各种药理肽，这些肽对癌细胞表面异常表达的离子通道具有选择性活性，可对这种疾病发挥强大的抗癌作用。
通过电刺激从菲律宾狼蛛中提取毒液，并通过反相高效液相色谱（RP-HPLC）进行分离。收集毒液馏分并用于体外分析，如细胞毒性、形态评估和氧化应激水平。
粗蜘蛛毒液的分离产生了几个峰，这些峰主要通过分光光度法和比色法检测为肽。几种蜘蛛毒液肽处理 MCF-7 细胞系会诱导几种内源性自由基的产生，如羟自由基（•OH）、一氧化氮自由基（•NO）、超氧阴离子自由基（•O2-）和丙二醛（MDA）反应产生的脂质过氧化物，这与 400 µg/mL 维生素 E 和 L-半胱氨酸提供的清除效果相当（p<0.05）。同时，产生的自由基通过罗丹明 123 和四唑染料分别降低线粒体膜电位和代谢活性（p>0.05）。这表现在 MCF-7 细胞中的细胞毒性增加，如细胞膜起泡、细胞脱落、半胱天冬酶活性和核片段化增加。
这些数据表明，菲律宾狼蛛毒液含有表现出对 MCF-7 细胞产生促氧化和亚硝化依赖的细胞毒性的肽成分，并可以提供其作为癌症治疗中的促氧化剂的潜在应用的机制见解。
。
