Bradshaw Michael J, Saviola Anthony J, Fesler Elizabeth, Mackessy Stephen P
School of Biological Sciences, University of Northern Colorado, 501 20th St., CB 92, Greeley, CO, 80639-0017, USA.
Cytotechnology. 2016 Aug;68(4):687-700. doi: 10.1007/s10616-014-9820-2. Epub 2014 Nov 19.
Snake venoms are mixtures of bioactive proteins and peptides that exhibit diverse biochemical activities. This wide array of pharmacologies associated with snake venoms has made them attractive sources for research into potentially novel therapeutics, and several venom-derived drugs are now in use. In the current study we performed a broad screen of a variety of venoms (61 taxa) from the major venomous snake families (Viperidae, Elapidae and "Colubridae") in order to examine cytotoxic effects toward MCF-7 breast cancer cells and A-375 melanoma cells. MTT cell viability assays of cancer cells incubated with crude venoms revealed that most venoms showed significant cytotoxicity. We further investigated venom from the Red-bellied Blacksnake (Pseudechis porphyriacus); venom was fractionated by ion exchange fast protein liquid chromatography and several cytotoxic components were isolated. SDS-PAGE and MALDI-TOF mass spectrometry were used to identify the compounds in this venom responsible for the cytotoxic effects. In general, viper venoms were potently cytotoxic, with MCF-7 cells showing greater sensitivity, while elapid and colubrid venoms were much less toxic; notable exceptions included the elapid genera Micrurus, Naja and Pseudechis, which were quite cytotoxic to both cell lines. However, venoms with the most potent cytotoxicity were often not those with low mouse LD50s, including some dangerously venomous viperids and Australian elapids. This study confirmed that many venoms contain cytotoxic compounds, including catalytic PLA2s, and several venoms also showed significant differential toxicity toward the two cancer cell lines. Our results indicate that several previously uncharacterized venoms could contain promising lead compounds for drug development.
蛇毒是具有多种生化活性的生物活性蛋白质和肽的混合物。与蛇毒相关的广泛药理作用使其成为研究潜在新型疗法的有吸引力的来源,目前有几种源自蛇毒的药物正在使用。在本研究中,我们对主要毒蛇科(蝰蛇科、眼镜蛇科和“游蛇科”)的多种毒液(61个分类单元)进行了广泛筛选,以检测其对MCF-7乳腺癌细胞和A-375黑色素瘤细胞的细胞毒性作用。用粗毒液孵育癌细胞的MTT细胞活力测定表明,大多数毒液显示出显著的细胞毒性。我们进一步研究了红腹黑蛇(Pseudechis porphyriacus)的毒液;通过离子交换快速蛋白质液相色谱对毒液进行分离,并分离出几种细胞毒性成分。使用SDS-PAGE和MALDI-TOF质谱法鉴定该毒液中负责细胞毒性作用的化合物。一般来说,蝰蛇毒液具有很强的细胞毒性,MCF-7细胞表现出更高的敏感性,而眼镜蛇科和游蛇科的毒液毒性要小得多;值得注意的例外包括属于眼镜蛇科的珊瑚蛇属、眼镜蛇属和红腹黑蛇属,它们对两种细胞系都有很强的细胞毒性。然而,细胞毒性最强的毒液往往不是那些小鼠半数致死量低的毒液,包括一些剧毒的蝰蛇科和澳大利亚眼镜蛇科。这项研究证实,许多毒液含有细胞毒性化合物,包括催化性磷脂酶A2,并且几种毒液对两种癌细胞系也表现出显著的毒性差异。我们的结果表明,几种以前未表征的毒液可能含有用于药物开发的有前景的先导化合物。