CD63 Tetraspanin Is a Negative Driver of Epithelial-to-Mesenchymal Transition in Human Melanoma Cells
CD63 Tetraspanin Is a Negative Driver of Epithelial-to-Mesenchymal Transition in Human Melanoma Cells
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DOI：
10.1038/jid.2014.258
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发表时间：
2014-12-01
影响因子：
6.5
通讯作者：
Lulli, Matteo
中科院分区：
文献类型：
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作者：
Lupia, Antonella;Peppicelli, Silvia;Lulli, Matteo
CD63四联蛋白在黑色素瘤的早期阶段高表达，在晚期病变中降低，表明它可能是肿瘤进展的抑制因子。我们采用基因功能缺失和基因功能获得的方法来研究CD63在黑色素瘤进展和上皮-间质转化（EMT）程序获得中的作用。我们使用了来自原发肿瘤的两种人类黑色素瘤细胞系和从皮肤转移中分离的一种原发人类黑色素瘤细胞系，它们的CD63表达水平不同。cd63沉默的黑色素瘤细胞表现出增强的运动性和侵袭性，E-cadherin下调，N-cadherin和Snail上调。在平行实验中，CD63的短暂和稳定的异位表达导致细胞活力、侵袭性和蛋白酶活性的显著降低，这与CD63蛋白水平的增加成正比。转染过表达最高水平CD63的细胞移植到免疫缺陷小鼠体内后，肿瘤的发病率和生长速度均有所降低。此外，这些细胞在基础条件和随后的TGF - β刺激下均表现出N-cadherin、Vimentin、Zeb1和a- sma的减少，并且对EMT程序有明显的抗性。因此，我们的研究结果在黑素瘤中建立了以前未报道的四跨蛋白CD63和EMT消除之间的机制联系。
The CD63 tetraspanin is highly expressed in the early stages of melanoma and decreases in advanced lesions, suggesting it as a possible suppressor of tumor progression. We employed loss- and gain-of-gene-function approaches to investigate the role of CD63 in melanoma progression and acquisition of the epithelial-to-mesenchymal transition (EMT) program. We used two human melanoma cell lines derived from primary tumors and one primary human melanoma cell line isolated from a cutaneous metastasis, differing by levels of CD63 expression. CD63-silenced melanoma cells showed enhanced motility and invasiveness with downregulation of E-cadherin and upregulation of N-cadherin and Snail. In parallel experiments, transient and stable ectopic expression of CD63 resulted in a robust reduction of cell motility, invasiveness, and protease activities, which was proportional to the increase in CD63 protein level. Transfected cells overexpressing the highest level of CD63 when transplanted into immunodeficient mice showed a reduced incidence and rate of tumor growth. Moreover, these cells showed a reduction of N-cadherin, Vimentin, Zeb1, and a-SMA, and a significant resistance to undergo an EMT program both in basal condition and in the following stimulation with TGF beta. Thus, our results establish a previously unreported mechanistic link between the tetraspanin CD63 and EMT abrogation in melanoma.