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二维差异凝胶电泳揭示钙网蛋白作为黏液样脂肪肉瘤中一种新型潜在转移相关蛋白

Calreticulin as A Novel Potential Metastasis-Associated Protein in Myxoid Liposarcoma, as Revealed by Two-Dimensional Difference Gel Electrophoresis.

作者信息

Tajima Takashi, Kito Fusako, Yoshida Akihiko, Kawai Akira, Kondo Tadashi

机构信息

Division of Rare Cancer Research, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan.

Department of Orthopaedic Surgery, Kyorin University School of Medicine, 6-20-2 Shinkawa, Mitaka, Tokyo 181-8611, Japan.

出版信息

Proteomes. 2019 Apr 10;7(2):13. doi: 10.3390/proteomes7020013.

Abstract

Myxoid liposarcoma (MLS) is a mesenchymal malignancy. To identify innovate seeds for clinical applications, we examined the proteomes of primary tumor tissues from 10 patients with MLS with different statuses of postoperative metastasis. The protein expression profiles of tumor tissues were created, and proteins with differential expression associated with postoperative metastasis were identified by two-dimensional difference gel electrophoresis (2D-DIGE) and mass spectrometry. The validation was performed using specific antibodies and in vitro analyses. Using 2D-DIGE, we observed 1726 protein species and identified proteins with unique expression levels in metastatic MLS. We focused on the overexpression of calreticulin in metastatic MLS. The higher expression of calreticulin was confirmed by Western blotting, and gene silencing assays demonstrated that reduced expression of calreticulin inhibited cell growth and invasion. Our findings suggested the important roles of calreticulin in MLS metastasis and supported its potential utility as a prognostic biomarker in MLS. Further investigations of the functional properties of calreticulin and other proteins identified in this study will improve our understanding of the biology of MLS and facilitate novel clinical applications.

摘要

黏液样脂肪肉瘤(MLS)是一种间叶组织恶性肿瘤。为了确定临床应用的创新靶点,我们检测了10例具有不同术后转移状态的MLS患者原发肿瘤组织的蛋白质组。构建了肿瘤组织的蛋白质表达谱,并通过二维差异凝胶电泳(2D-DIGE)和质谱鉴定了与术后转移相关的差异表达蛋白质。使用特异性抗体和体外分析进行验证。通过2D-DIGE,我们观察到1726种蛋白质,并鉴定出转移性MLS中具有独特表达水平的蛋白质。我们重点关注了钙网蛋白在转移性MLS中的过表达。蛋白质免疫印迹法证实了钙网蛋白的高表达,基因沉默实验表明钙网蛋白表达降低会抑制细胞生长和侵袭。我们的研究结果表明钙网蛋白在MLS转移中起重要作用,并支持其作为MLS预后生物标志物的潜在用途。对本研究中鉴定的钙网蛋白和其他蛋白质的功能特性进行进一步研究,将增进我们对MLS生物学的理解,并促进新的临床应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f402/6631384/23be16d83d6c/proteomes-07-00013-g001.jpg

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