Cancer Prevention Basic Research Project, National Cancer Center Research Institute, Tokyo, Japan .
Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Tokyo, Japan .
Proc Jpn Acad Ser B Phys Biol Sci. 2007 Sep;83(6):175-8. doi: 10.2183/pjab.83.175.
Crude extracts from larvae, pupae and adults of cabbage white butterflies, Pieris rapae and Pieris brassicae, and green-veined butterfly, Pieris napi, have an ability to induce apoptosis in the human cancer cell lines. As apoptosis inducing protein, pierisin-1 and -2 have been isolated from pupae of P. rapae and P. brassicae, respectively, and shown to exhibit DNA ADP-ribosylating activity. Although the highest activity was detected in the late phase of larvae and early phase of pupae, certain activity was found in adult butterflies. In order to investigate distribution of substances having pierisin-like activities in butterflies, many species need to be analyzed. However, fresh samples of larvae and pupae are hard to obtain, especially if samples are of scarce species or from overseas. The usage of adult butterflies is practical to examine the distribution of pierisin-like activity in many species. In this study, we examined the cytotoxicity of crude extracts from adults of P. rapae against HeLa cells and DNA ADP-ribosylation ability during storage for 1, 2 and 8 weeks at room temperature after killing adult butterflies after eclosion. Body weights decreased to 18% for 8 weeks through dehydration. Cytotoxicity of samples from butterfly kept for 1, 2 and 8 weeks decreased to 47, 39 and 22%, respectively, of the control value. DNA ADP-ribosylating activity of the samples also decreased to 30, 27 and 23%. Similar reduction was observed on western blot analysis with anti-pierisin-1 antibody. Fortunately, these results suggest that cytotoxic and DNA ADP-ribosylating activity persists to some extent in the body after killing, at least for 8 weeks. Thus, butterfly adult samples kept for two months at room temperature can still be useful for examination of the presence of substance having pierisin-like activity.
菜粉蝶、菜青虫和甘蓝粉蝶幼虫、蛹和成虫以及绿斑蛱蝶粗提物具有诱导人癌细胞凋亡的能力。作为凋亡诱导蛋白，分别从菜粉蝶和菜青虫的蛹中分离出了 pierisin-1 和 -2，并显示出具有 DNA ADP-ribosylating 活性。尽管在幼虫后期和蛹早期检测到最高活性，但在成蝶中也发现了一定的活性。为了研究具有 pierisin 样活性的物质在蝴蝶中的分布，需要对许多物种进行分析。然而，幼虫和蛹的新鲜样本很难获得，特别是对于稀有物种或来自海外的样本。使用成蝶来检验许多物种中 pierisin 样活性的分布是很实用的。在这项研究中，我们检测了刚羽化的成蝶死后，室温下储存 1、2 和 8 周时，从菜粉蝶成虫粗提取物对 HeLa 细胞的细胞毒性和 DNA ADP-ribosylation 能力。8 周后，成虫因脱水体重降至 18%。储存 1、2 和 8 周的样本的细胞毒性分别降至对照值的 47%、39%和 22%。样本的 DNA ADP-ribosylating 活性也分别下降至 30%、27%和 23%。用抗 pierisin-1 抗体进行的 western blot 分析也观察到了类似的减少。幸运的是，这些结果表明，在杀死后，至少在 8 周内，成虫体内仍具有一定程度的细胞毒性和 DNA ADP-ribosylating 活性。因此，在室温下储存两个月的蝴蝶成虫样本仍然可用于检查是否存在具有 pierisin 样活性的物质。
