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无标记定量蛋白质组学鉴定的 PDLIM5 作为甲状腺乳头状癌潜在的新型生物标志物。

PDLIM5 identified by label-free quantitative proteomics as a potential novel biomarker of papillary thyroid carcinoma.

机构信息

Molecular Medicine and Cancer Research Center, Chongqing Medical University, Yuzhong District, No.1, Medical Road, Chongqing 400016, China; Medicine and Pharmacy Research Center, Binzhou Medical University, Laishan District, No.346, Guanhai Road, Yantai, Shandong Province 264003, China.

Medicine and Pharmacy Research Center, Binzhou Medical University, Laishan District, No.346, Guanhai Road, Yantai, Shandong Province 264003, China.

出版信息

Biochem Biophys Res Commun. 2018 May 5;499(2):338-344. doi: 10.1016/j.bbrc.2018.03.159. Epub 2018 Mar 24.

Abstract

In order to better understand the mechanisms underlying the development of papillary thyroid carcinoma (PTC), and to identify new potential biomarkers, high-resolution label-free mass spectrometry was performed on PTC tissues and adjacent normal thyroid tissues from six patients. In this process, 2788 proteins were identified, out of which 49 proteins presented significant differences between PTC tissues and adjacent normal thyroid tissues. Gene ontology revealed that the majority of these proteins are involved in the catalytic activity and binding. We selected three proteins with differential expressions: PDZ and LIM domain 5 (PDLIM5), PDLIM1 and ALDH1A1; Protein expressions were further verified by RT-PCR and western blot. Among these, expression of PDLIM5 and PDLIM1 was up-regulated, while that of ALDH1A1 was down-regulated in PTC tissues. Next, we confirmed their expression through quantitative dot blot (QDB) technique. We found that knockdown of PDLIM5 expression in the B-CPAP cell line could inhibit the migration, invasion and proliferation of PTC cells. In addition, PDLIM5 knockdown reduced Ras and Phospho-ERK1/2 expression. Thus, we suggested that PDLIM5 promotes PTC via activation of the Ras-ERK pathway. Our research provides new molecular insight into the function of PDLIM5, which may assist in studying the mechanism of PTC. In addition, PDLIM5 could be further explored as a potential candidate for PTC treatment.

摘要

为了更好地理解甲状腺乳头状癌(PTC)发生发展的机制,寻找新的潜在的生物标志物,我们对 6 例 PTC 组织及其相邻正常甲状腺组织进行了高分辨率无标记质谱分析。在这个过程中,鉴定到了 2788 种蛋白质,其中 49 种蛋白质在 PTC 组织和相邻正常甲状腺组织之间存在显著差异。GO 分析显示,这些蛋白质主要参与催化活性和结合。我们选择了 3 种具有差异表达的蛋白质:PDZ 和 LIM 结构域蛋白 5(PDLIM5)、PDLIM1 和 ALDH1A1;通过 RT-PCR 和 Western blot 进一步验证了它们的蛋白表达。其中,PDLIM5 和 PDLIM1 的表达在 PTC 组织中上调,而 ALDH1A1 的表达下调。接下来,我们通过定量斑点印迹(QDB)技术验证了它们的表达。我们发现,在 B-CPAP 细胞系中敲低 PDLIM5 的表达可以抑制 PTC 细胞的迁移、侵袭和增殖。此外,PDLIM5 敲低降低了 Ras 和 Phospho-ERK1/2 的表达。因此,我们认为 PDLIM5 通过激活 Ras-ERK 通路促进 PTC 的发生。我们的研究为 PDLIM5 的功能提供了新的分子见解,可能有助于研究 PTC 的发病机制。此外,PDLIM5 可以进一步探索作为 PTC 治疗的潜在候选物。

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