Regulation of viability, differentiation and death of human melanoma cells carrying neural stem cell biomarkers: a possibility for neural trans-differentiation
Regulation of viability, differentiation and death of human melanoma cells carrying neural stem cell biomarkers: a possibility for neural trans-differentiation
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DOI：
10.1007/s10495-015-1131-3
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发表时间：
2015-07-01
期刊：
影响因子：
7.2
通讯作者：
Hei, Tom K.
中科院分区：
文献类型：
--
作者：
Ivanov, Vladimir N.;Hei, Tom K.
在胚胎发育过程中，黑素细胞的前体--成黑素细胞从神经嵴干细胞的亚群中出现，并迁移到皮肤上。黑色素瘤在成黑素细胞分化成黑素细胞期间产生，并且通过体细胞诱变和表观遗传调节从年轻的增殖黑素细胞产生。在本研究中，我们使用了几个人黑色素瘤细胞系，从黑色素瘤发展的连续阶段（径向生长期、垂直生长期和转移期）比较：（i）通过凋亡和坏死性凋亡诱导细胞死亡的频率和效率;（ii）黑素瘤细胞的贴壁培养物和球状体培养物中神经和癌症干细胞生物标志物以及死亡受体DR 5和FAS的存在;（iii）内源性细胞因子产生的抗凋亡作用和（iv）黑素瘤细胞进行神经转分化的能力。我们证明，程序性坏死或坏死性凋亡，可以诱导两个转移性黑色素瘤系，FEMX和OM 431，而细胞凋亡的线粒体途径是在绝大多数黑色素瘤系普遍存在。本研究中使用的所有黑色素瘤细胞系均表达大量多能性标志物SOX 2和NANOG。在黑色素瘤进展过程中，早期神经祖细胞标志物巢蛋白的表达有增加的趋势。大多数黑色素瘤细胞系，包括WM 35，FEMX和A375，可以在含有补充剂的无血清培养基中生长为球状体培养物。在添加胰岛素的无血清培养基中，1205 Lu和OM 431黑色素瘤细胞可以诱导神经转分化。这通过神经元标记物doublecortin和β 3-Tubulin的表达、神经突的显著生长以及显性阴性Rac 1 N17对该过程的负调节来证实。这些结果表明，分化的黑色素瘤细胞的相对可塑性和它们的神经转分化的可能性，而不需要初步的去分化。
During embryonic development, melanoblasts, the precursors of melanocytes, emerge from a subpopulation of the neural crest stem cells and migrate to colonize skin. Melanomas arise during melanoblast differentiation into melanocytes and from young proliferating melanocytes through somatic mutagenesis and epigenetic regulations. In the present study, we used several human melanoma cell lines from the sequential phases of melanoma development (radial growth phase, vertical growth phase and metastatic phase) to compare: (i) the frequency and efficiency of the induction of cell death via apoptosis and necroptosis; (ii) the presence of neural and cancer stem cell biomarkers as well as death receptors, DR5 and FAS, in both adherent and spheroid cultures of melanoma cells; (iii) anti-apoptotic effects of the endogenous production of cytokines and (iv) the ability of melanoma cells to perform neural trans-differentiation. We demonstrated that programed necrosis or necroptosis, could be induced in two metastatic melanoma lines, FEMX and OM431, while the mitochondrial pathway of apoptosis was prevalent in a vast majority of melanoma lines. All melanoma lines used in the current study expressed substantial levels of pluripotency markers, SOX2 and NANOG. There was a trend for increasing expression of Nestin, an early neuroprogenitor marker, during melanoma progression. Most of the melanoma lines, including WM35, FEMX and A375, can grow as a spheroid culture in serum-free media with supplements. It was possible to induce neural trans-differentiation of 1205Lu and OM431 melanoma cells in serum-free media supplemented with insulin. This was confirmed by the expression of neuronal markers, doublecortin and beta 3-Tubulin, by significant growth of neurites and by the negative regulation of this process by a dominant-negative Rac1N17. These results suggest a relative plasticity of differentiated melanoma cells and a possibility for their neural trans-differentiation without the necessity for preliminary dedifferentiation.