Shimura Mitsuhiro, Mizuma Masamichi, Takadate Tatsuyuki, Katoh Yasutake, Suzuki Takashi, Iseki Masahiro, Hata Tatsuo, Aoki Shuichi, Suzuki Yukie, Sakata Naoaki, Ohtsuka Hideo, Hayashi Hiroki, Morikawa Takanori, Nakagawa Kei, Motoi Fuyuhiko, Naitoh Takeshi, Igarashi Kazuhiko, Sasano Hironobu, Unno Michiaki
Department of Surgery, Tohoku University Graduate School of Medicine, Aobaku, Sendai 980-8574, Japan.
Department of Biochemistry, Tohoku University Graduate School of Medicine, Aobaku, Sendai 980-8574, Japan.
Oncotarget. 2018 May 11;9(36):24291-24303. doi: 10.18632/oncotarget.25110.
The aim of this study was to identify novel liver metastasis-correlated proteins of PanNEN by proteomics to compare pancreatic tumor (PT) with paired metastatic liver tumor (LT). Of 118 surgical cases with PanNEN, 7 cases with formalin-fixed paraffin-embedded (FFPE) tissues of both PT and paired LT were evaluated by proteomics. Tumor cells were selectively collected from FFPE tissues by laser capture microdissection. A total of 3,722 proteins were detected from extracted peptides by mass spectrometry-based shotgun analysis. Selection of the candidate proteins expressed differently between PT and LT were performed by semi-quantitative comparison and confirmation with immunohistochemistry. We focused on ANXA6, CNPY2, RAB11B and TUBB3, all of which had higher expressions in LT. In all surgical cases with FFPE samples, liver recurrence-free survival (RFS) was evaluated in correlation to the expression of the candidate proteins in PT by immunohistochemistry. Liver RFS was significantly poorer in CNPY2 positive patients than in negative patients (10-year liver RFS; 39.8% vs. 92.3%, p = 0.012). Also, liver RFS tended to be poorer in ANXA6 positive patients than in those who were negative (10-year liver RFS; 51.4% vs. 95.0%, p = 0.099). In the multivariate analysis, the independent predictors of liver RFS were CNPY2 positivity (HR: 6.19, 95 % CI: 1.47-42.79, p = 0.011) and tumor size ≥ 42 mm (HR: 4.63, 95 % CI: 1.03-23.23, p = 0.045). In conclusion, CNPY2 is a novel liver metastasis-correlated protein of PanNEN.
本研究旨在通过蛋白质组学鉴定与胰腺神经内分泌肿瘤（PanNEN）肝转移相关的新蛋白，以比较胰腺肿瘤（PT）与配对的肝转移肿瘤（LT）。在118例PanNEN手术病例中，对7例同时具有PT和配对LT的福尔马林固定石蜡包埋（FFPE）组织的病例进行了蛋白质组学评估。通过激光捕获显微切割从FFPE组织中选择性收集肿瘤细胞。通过基于质谱的鸟枪法分析从提取的肽中检测到总共3722种蛋白质。通过半定量比较和免疫组织化学确认来选择PT和LT之间表达不同的候选蛋白质。我们重点关注膜联蛋白A6（ANXA6）、卷曲霉素样蛋白2（CNPY2）、RAB11B和微管蛋白β3（TUBB3），所有这些蛋白在LT中均有较高表达。在所有具有FFPE样本的手术病例中，通过免疫组织化学评估肝无复发生存期（RFS）与PT中候选蛋白表达的相关性。CNPY2阳性患者的肝RFS明显比阴性患者差（10年肝RFS；39.8%对92.3%，p = 0.012）。此外，ANXA6阳性患者的肝RFS也倾向于比阴性患者差（10年肝RFS；51.4%对95.0%，p = 0.099）。在多变量分析中，肝RFS的独立预测因素是CNPY2阳性（风险比：6.19，95%置信区间：1.47 - 42.79，p = 0.011）和肿瘤大小≥42 mm（风险比：4.63，95%置信区间：1.03 - 23.23，p = 0.045）。总之，CNPY2是一种与PanNEN肝转移相关的新蛋白。
