Department of Pharmacology & Chemical Biology, University of Pittsburgh Cancer Institute, University of Pittsburgh, PA, USA.
J Proteome Res. 2010 Jul 2;9(7):3656-63. doi: 10.1021/pr100164x.
Gaining insights into the molecular events that govern the progression from melanoma in situ to advanced melanoma and understanding how the local microenvironment at the melanoma site influences this progression are two clinically pivotal aspects that to date are largely unexplored. In an effort to identify key regulators of the crosstalk between melanoma cells and the melanoma-skin microenvironment, primary and metastatic human melanoma cells were seeded into skin organ cultures (SOCs) and grown for two weeks. Melanoma cells were recovered from SOCs by laser microdissection and whole-cell tryptic digests were analyzed by nanoflow liquid chromatography-tandem mass spectrometry. The differential protein abundances were calculated by spectral counting, the results of which provides evidence that cell-matrix and cell-adhesion molecules that are upregulated in the presence of these melanoma cells recapitulate proteomic data obtained from comparative analysis of human biopsies of invasive melanoma and a tissue sample of adjacent, noninvolved skin. This concordance demonstrates the value of SOCs for conducting proteomic investigations of the melanoma microenvironment.
深入了解从原位黑色素瘤进展为晚期黑色素瘤的分子事件，并理解黑色素瘤部位的局部微环境如何影响这种进展，是两个具有重要临床意义但目前尚未得到充分探索的方面。为了鉴定黑色素瘤细胞与黑色素瘤皮肤微环境之间相互作用的关键调节因子，将原发性和转移性人类黑色素瘤细胞接种到皮肤器官培养物（SOC）中并培养两周。通过激光微切割从 SOC 中回收黑色素瘤细胞，并用纳流液相色谱-串联质谱法分析全细胞胰蛋白酶消化物。通过光谱计数计算差异蛋白丰度，结果表明，在这些黑色素瘤细胞存在的情况下上调的细胞-基质和细胞黏附分子重现了从侵袭性黑色素瘤患者活检和相邻未受累皮肤组织样本的比较分析中获得的蛋白质组学数据。这种一致性证明了 SOC 用于进行黑色素瘤微环境蛋白质组学研究的价值。
