Byström Sanna, Fredolini Claudia, Edqvist Per-Henrik, Nyaiesh Etienne-Nicholas, Drobin Kimi, Uhlén Mathias, Bergqvist Michael, Pontén Fredrik, Schwenk Jochen M
Affinity Proteomics, SciLifeLab, KTH - Royal Institute of Technology, 171 65 Solna, Sweden.
Department of Immunology, Genetics and Pathology, Uppsala University, 751 85 Uppsala, Sweden; Science for Life Laboratory, Uppsala University, 751 85 Uppsala, Sweden.
Transl Oncol. 2017 Jun;10(3):385-395. doi: 10.1016/j.tranon.2017.03.002. Epub 2017 Apr 20.
Blood-based proteomic profiling may aid and expand our understanding of diseases and their different phenotypes. The aim of the presented study was to profile serum samples from patients with malignant melanoma using affinity proteomic assays to describe proteins in the blood stream that are associated to stage or recurrence of melanoma.
Multiplexed protein analysis was conducted using antibody suspension bead arrays. A total of 232 antibodies against 132 proteins were selected from (i) a screening with 4595 antibodies and 32 serum samples from melanoma patients and controls, (ii) antibodies used for immunohistochemistry, (iii) protein targets previously related with melanoma. The analysis was performed with 149 serum samples from patients with malignant melanoma. Antibody selectivity was then assessed by Western blot, immunocapture mass spectrometry, and epitope mapping. Lastly, indicative antibodies were applied for IHC analysis of melanoma tissues.
Serum levels of regucalcin (RGN) and syntaxin 7 (STX7) were found to be lower in patients with both recurring tumors and a high Breslow's thickness (T-stage 3/4) compared to low thickness (T-stage 1/2) without disease recurrence. Serum levels of methylenetetrahydrofolate dehydrogenase 1-like (MTHFD1L) were instead elevated in sera of T3/4 patients with recurrence. The analysis of tissue sections with S100A6 and MTHFD1L showed positive staining in a majority of patients with melanoma, and S100A6 was significantly associated to T-stage.
Our findings provide a starting point to further study RGN, STX7, MTHFD1L and S100A6 in serum to elucidate their involvement in melanoma progression and to assess a possible contribution to support clinical indications.
基于血液的蛋白质组学分析有助于增进并拓展我们对疾病及其不同表型的理解。本研究旨在利用亲和蛋白质组学分析对恶性黑色素瘤患者的血清样本进行分析，以描述血流中与黑色素瘤分期或复发相关的蛋白质。
使用抗体悬浮珠阵列进行多重蛋白质分析。从以下方面选择了针对132种蛋白质的总共232种抗体：（i）对4595种抗体和来自黑色素瘤患者及对照的32份血清样本进行的筛选；（ii）用于免疫组织化学的抗体；（iii）先前与黑色素瘤相关的蛋白质靶点。对149份恶性黑色素瘤患者的血清样本进行了分析。然后通过蛋白质印迹、免疫捕获质谱和表位作图评估抗体的选择性。最后，将指示性抗体应用于黑色素瘤组织的免疫组织化学分析。
与厚度较低（T分期1/2）且无疾病复发的患者相比，复发肿瘤且Breslow厚度较高（T分期3/4）的患者血清中调钙蛋白（RGN）和Syntaxin 7（STX7）水平较低。相反，复发的T3/4期患者血清中亚甲基四氢叶酸脱氢酶1样蛋白（MTHFD1L）水平升高。对S100A6和MTHFD1L组织切片的分析显示，大多数黑色素瘤患者呈阳性染色，且S100A6与T分期显著相关。
我们的研究结果为进一步研究血清中的RGN、STX7、MTHFD1L和S100A6提供了一个起点，以阐明它们在黑色素瘤进展中的作用，并评估其对支持临床指征的可能贡献。
