Kodet Ondřej, Dvořánková Barbora, Krejčí Eliška, Szabo Pavol, Dvořák Petr, Štork Jiří, Krajsová Ivana, Dundr Pavel, Smetana Karel, Lacina Lukáš
1st Faculty of Medicine, Department of Dermatovenereology, Charles University, Prague, Czech Republic.
Tumour Biol. 2013 Dec;34(6):3345-55. doi: 10.1007/s13277-013-0905-x. Epub 2013 Jun 12.
Malignant melanoma is a highly aggressive tumor with increasing incidence and high mortality. The importance of immunohistochemistry in diagnosis of the primary tumor and in early identification of metastases in lymphatic nodes is enormous; however melanoma phenotype is frequently variable and thus several markers must be employed simultaneously. The purposes of this study are to describe changes of phenotype of malignant melanoma in vitro and in vivo and to investigate whether changes of environmental factors mimicking natural conditions affect the phenotype of melanoma cells and can revert the typical in vitro loss of diagnostic markers. The influence of microenvironment was studied by means of immunocytochemistry on co-cultures of melanoma cells with melanoma-associated fibroblast and/or in conditioned media. The markers typical for melanoma (HMB45, Melan-A, Tyrosinase) were lost in malignant cells isolated from malignant effusion; however, tumor metastases shared identical phenotype with primary tumor (all markers positive). The melanoma cell lines also exerted reduced phenotype in vitro. The only constantly present diagnostic marker observed in our experiment was S100 protein and, in lesser extent, also Nestin. The phenotype loss was reverted under the influence of melanoma-associated fibroblast and/or both types of conditioned media. Loss of some markers of melanoma cell phenotype is not only of diagnostic significance, but it can presumably also contribute to biological behavior of melanoma. The presented study shows how the conditions of cultivation of melanoma cells can influence their phenotype. This observation can have some impact on considerations about the role of microenvironment in tumor biology.
恶性黑色素瘤是一种侵袭性很强的肿瘤，发病率不断上升且死亡率很高。免疫组织化学在原发性肿瘤诊断及早期识别淋巴结转移方面的重要性极大；然而，黑色素瘤的表型常常多变，因此必须同时使用多种标志物。本研究的目的是描述恶性黑色素瘤在体外和体内的表型变化，并探究模拟自然条件的环境因素变化是否会影响黑色素瘤细胞的表型，以及能否逆转体外典型的诊断标志物缺失情况。通过免疫细胞化学方法，研究黑色素瘤细胞与黑色素瘤相关成纤维细胞共培养物及/或条件培养基中微环境的影响。从恶性积液中分离出的恶性细胞中，黑色素瘤典型的标志物（HMB45、Melan-A、酪氨酸酶）缺失；然而，肿瘤转移灶与原发性肿瘤具有相同的表型（所有标志物均为阳性）。黑色素瘤细胞系在体外也表现出表型减弱。在我们的实验中观察到唯一持续存在的诊断标志物是S100蛋白，其次是巢蛋白。在黑色素瘤相关成纤维细胞和/或两种类型的条件培养基的影响下，表型缺失得以逆转。黑色素瘤细胞表型某些标志物的缺失不仅具有诊断意义，还可能对黑色素瘤的生物学行为有影响。本研究展示了黑色素瘤细胞的培养条件如何影响其表型。这一观察结果可能会对微环境在肿瘤生物学中的作用的相关思考产生一定影响。
