Walker Charles, Böttger S Anne, Mulkern Joseph, Jerszyk Emily, Litvaitis Marianne, Lesser Michael
Department of Molecular, Cellular and Biomedical Sciences, Rudman Hall, University of New Hampshire, Durham, New Hampshire 03824, USA.
Biol Bull. 2009 Feb;216(1):23-39. doi: 10.1086/BBLv216n1p23.
On the northeastern coast of the United States and Canada, Mya arenaria, the soft shell clam, develops a diffuse, hemopoetic tumor (a fatal leukemia-like cancer) resulting from inactivation of p53-like family member proteins.These malignant cells provide a model for an unrelated set of human cancer cells that are also characterized by mortalin-based cytoplasmic sequestration of wild-type p53 tumor suppressor protein (mortalin is the mitochondrial Hsp70 protein). Here we describe methods for mass culture and long-term storage of tumor cells from this cancer. These are the first successful efforts at maintaining malignant cells from any marine invertebrate in vitro. Following passage (subculture), these cultures undergo transition from primary cultures to non-immortalized cell lines that continue to proliferate and do not re-differentiate the normal hemocyte phenotype. We also characterize normal clam hemocytes and the pathology of cancerous clam hemocytes in vitro and in vivo using light and electron microscopy, cyto- and immunocytochemistry, molecular biology, and a phagocytosis assay. Our protocols provide biomedical and environmental researchers with ready access to this naturally occurring cancer model. We discuss the clam cancer model regarding (a) human health and disease; (b) animal health, disease, and aquaculture; (c) environmental health monitoring; and (d) future research directions.
在美国和加拿大的东北海岸，软壳蛤（Mya arenaria）会患上一种弥漫性的造血肿瘤（一种致命的白血病样癌症），这是由p53样家族成员蛋白失活导致的。这些恶性细胞为一组不相关的人类癌细胞提供了一个模型，这些人类癌细胞的特征也是野生型p53肿瘤抑制蛋白基于mortalin的细胞质隔离（mortalin是线粒体热休克蛋白70）。在这里，我们描述了从这种癌症中大规模培养和长期保存肿瘤细胞的方法。这是首次成功地在体外维持任何海洋无脊椎动物的恶性细胞。传代（继代培养）后，这些培养物从原代培养转变为非永生化细胞系，这些细胞系继续增殖，不会重新分化为正常血细胞表型。我们还使用光学和电子显微镜、细胞和免疫细胞化学、分子生物学以及吞噬试验，在体外和体内对正常蛤血细胞以及癌性蛤血细胞的病理学进行了表征。我们的方案为生物医学和环境研究人员提供了使用这个天然存在的癌症模型的便利途径。我们讨论了蛤癌症模型在以下方面的情况：（a）人类健康与疾病；（b）动物健康、疾病与水产养殖；（c）环境卫生监测；以及（d）未来的研究方向。
