Attarde Saurabh S, Pandit Sangeeta V
Department of Zoology, Savitribai Phule Pune University, Ganeshkhind, Pune, Maharashtra, 411007, India.
BMC Complement Altern Med. 2017 Nov 28;17(1):503. doi: 10.1186/s12906-017-2018-3.
Breast cancer is the most common cancer which causes significant morbidity and mortality among women worldwide. Lack of medical facilities for early detection, therapeutic strategies for treatment and side effects due to pharmacological compounds have encompassed the need for new therapies mostly from natural sources. A lot of components have been identified from different snake venoms as therapeutic agents. A group of polypeptides (60-70 amino acid residues) called cytotoxins or cardiotoxins present in an elapid family of snakes have a wide variety of pharmaceutical actions and have the tendency to damage a wide variety of cells including cancerous cells. The aim of the present study was to evaluate the cytotoxic effect of NN-32 protein toxin purified from Indian Spectacled Cobra venom against human breast cancer cell lines (MCF-7 and MDA-MB-231).
The NN-32 toxin was purified by ion exchange chromatography and further by RP-HPLC. The potential anticancer effects of the NN-32 toxin on MCF-7 and MDA-MB-231 cells were evaluated using MTT, anti-proliferation, neutral red (NR) uptake and Lactate Dehydrogenase (LDH) release assay.
The ion exchange chromatography showed various peaks among fraction no. 35 showing cytotoxic activity and this fraction showed a single peak with retention time 3.6 mins by HPLC using C18 column. The NN-32 toxin induced cytotoxicity in MCF-7 and MDA-MB-231 cells with the IC value of 2.5 and 6.7 μg/ml respectively. The NN-32 showed significant cytotoxicity to both the cell lines along with low cytotoxicity to MCF-10A (normal breast epithelial) cells. The cytotoxic effect was further confirmed by the anti-proliferative, NR uptake and LDH release assays.
The purified toxin NN-32 from Naja naja venom showed cytotoxic activity against MCF-7 (ER+) and MDA-MB-231(ER-) cells in both dose dependent and time dependent manner.
乳腺癌是全球女性中最常见的癌症，会导致严重的发病率和死亡率。缺乏早期检测的医疗设施、治疗策略以及药物化合物的副作用，使得人们对主要来自天然来源的新疗法的需求不断增加。已从不同蛇毒中鉴定出许多成分作为治疗剂。眼镜蛇科蛇类中存在的一组称为细胞毒素或心脏毒素的多肽（60 - 70个氨基酸残基）具有广泛的药理作用，并且有损伤包括癌细胞在内的多种细胞的倾向。本研究的目的是评估从印度眼镜蛇毒液中纯化的NN - 32蛋白毒素对人乳腺癌细胞系（MCF - 7和MDA - MB - 231）的细胞毒性作用。
通过离子交换色谱法进一步通过反相高效液相色谱法纯化NN - 32毒素。使用MTT、抗增殖、中性红（NR）摄取和乳酸脱氢酶（LDH）释放试验评估NN - 32毒素对MCF - 7和MDA - MB - 231细胞的潜在抗癌作用。
离子交换色谱法在第35号馏分中显示出多个峰，该馏分具有细胞毒性活性，通过使用C18柱的HPLC分析，该馏分在保留时间3.6分钟时显示出单峰。NN - 32毒素在MCF - 7和MDA - MB - 231细胞中诱导细胞毒性，IC值分别为2.5和6.7μg/ml。NN - 32对两种细胞系均显示出显著的细胞毒性，而对MCF - 10A（正常乳腺上皮）细胞的细胞毒性较低。抗增殖、NR摄取和LDH释放试验进一步证实了细胞毒性作用。
从眼镜蛇毒液中纯化的毒素NN - 32对MCF - 7（雌激素受体阳性）和MDA - MB - 231（雌激素受体阴性）细胞显示出剂量依赖性和时间依赖性的细胞毒性活性。
