Jobe Njainday Pulo, Živicová Veronika, Mifková Alžběta, Rösel Daniel, Dvořánková Barbora, Kodet Ondřej, Strnad Hynek, Kolář Michal, Šedo Aleksi, Smetana Karel, Strnadová Karolina, Brábek Jan, Lacina Lukáš
Department of Cell Biology, Faculty of Sciences, Charles University in Prague, Viničná 7, 120 00, Prague 2, Czech Republic.
Biotechnology and Biomedicine Center of the Academy of Sciences and Charles University in Vestec (BIOCEV), Průmyslová 595, Vestec u Prahy, Prague, Czech Republic.
Histochem Cell Biol. 2018 May;149(5):503-516. doi: 10.1007/s00418-018-1650-4. Epub 2018 Feb 12.
Melanoma represents a malignant disease with steadily increasing incidence. UV-irradiation is a recognized key factor in melanoma initiation. Therefore, the efficient prevention of UV tissue damage bears a critical potential for melanoma prevention. In this study, we tested the effect of UV irradiation of normal keratinocytes and their consequent interaction with normal and cancer-associated fibroblasts isolated from melanoma, respectively. Using this model of UV influenced microenvironment, we measured melanoma cell migration in 3-D collagen gels. These interactions were studied using DNA microarray technology, immunofluorescence staining, single cell electrophoresis assay, viability (dead/life) cell detection methods, and migration analysis. We observed that three 10 mJ/cm fractions at equal intervals over 72 h applied on keratinocytes lead to a 50% increase (p < 0.05) in in vitro invasion of melanoma cells. The introduction cancer-associated fibroblasts to such model further significantly stimulated melanoma cells in vitro invasiveness to a higher extent than normal fibroblasts. A panel of candidate gene products responsible for facilitation of melanoma cells invasion was defined with emphasis on IL-6, IL-8, and CXCL-1. In conclusion, this study demonstrates a synergistic effect between cancer microenvironment and UV irradiation in melanoma invasiveness under in vitro condition.
黑色素瘤是一种发病率持续上升的恶性疾病。紫外线照射是黑色素瘤发病的一个公认关键因素。因此，有效预防紫外线对组织的损伤对预防黑色素瘤具有至关重要的潜在意义。在本研究中，我们分别测试了紫外线照射正常角质形成细胞的效果，以及随后它们与从黑色素瘤中分离出的正常成纤维细胞和癌相关成纤维细胞的相互作用。利用这种受紫外线影响的微环境模型，我们测量了黑色素瘤细胞在三维胶原凝胶中的迁移情况。通过DNA微阵列技术、免疫荧光染色、单细胞电泳分析、细胞活力（死/活）检测方法和迁移分析对这些相互作用进行了研究。我们观察到，在72小时内等间隔施加三次10 mJ/cm²的紫外线照射可使黑色素瘤细胞的体外侵袭能力提高50%（p < 0.05）。将癌相关成纤维细胞引入该模型进一步显著刺激了黑色素瘤细胞的体外侵袭能力，且刺激程度高于正常成纤维细胞。确定了一组有助于黑色素瘤细胞侵袭的候选基因产物，重点是白细胞介素-6、白细胞介素-8和CXC趋化因子配体-1。总之，本研究证明了在体外条件下，癌症微环境与紫外线照射在黑色素瘤侵袭方面存在协同效应。
