Up-regulation of PCOLCE by TWIST1 promotes metastasis in Osteosarcoma
Up-regulation of PCOLCE by TWIST1 promotes metastasis in Osteosarcoma
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TWIST1上调PCOLCE促进骨肉瘤转移
DOI：
10.7150/thno.34090
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发表时间：
2019-01-01
期刊：
影响因子：
12.4
通讯作者：
Kang, Tiebang
中科院分区：
文献类型：
--
作者：
Wang, Shang;Zhong, Li;Kang, Tiebang
前胶原蛋白C-蛋白酶增强子蛋白（PCOLCE）最初被鉴定为促进BMP 1催化前胶原的增强子。PCOLCE参与细胞外和角膜修复的重建。血液中PCOLCE的升高表明乳腺癌已经转移到骨骼中。然而，对PCOLCE的直接研究尚未报道。方法：通过RNA-seq分析从4例正常组织和16例骨肉瘤组织中鉴定ECM候选物。Transwell法检测骨肉瘤细胞的体外迁移和侵袭能力。建立自发转移性骨肉瘤模型，以评估骨肉瘤在体内的转移。通过PNGase F处理结合Western印迹鉴定N-连接的糖基化氨基酸。通过荧光素酶、qPCR和ChIP分析阐明了TWIST 1调控PCOLCE转录的机制。结果如下：PCOLCE在人骨肉瘤组织中的表达明显高于其在癌旁组织中的表达;组织中PCOLCE的高表达与患者预后不良相关，并且通过shRNA敲低PCOLCE会损害骨肉瘤细胞的迁移、侵袭和肺转移。野生型PCOLCE的过表达，而不是其N29 Q突变体，促进迁移，侵袭和转移，表明在Asn 29的糖基化的PCOLCE是必要的，其在骨肉瘤中的功能。TWIST 1是骨肉瘤转移的关键转录因子，也在骨肉瘤组织中过表达，并与PCOLCE或其潜在的前胶原底物（如COL 1A 1、COL 1A 2、COL 5A 1、COL 8A 2和COL 10A 1）呈正相关。结论：我们的研究结果是第一个提供证据，PCOLCE在促进骨肉瘤肺转移中起着关键作用，并且TWIST 1上调PCOLCE可能导致治疗转移性骨肉瘤患者的新治疗策略。
Procollagen C-proteinase enhancer protein (PCOLCE) was originally identified as an enhancer to facilitate the catalysis of procollagens by BMP1. PCOLCE participates in the reconstitution of extracellular and corneal repair. The elevation of PCOLCE in blood indicates that breast cancer has metastasized into the bones. However, direct research on PCOLCE has not been reported. Methods: ECM candidates were identified by RNA-seq analysis from 4 normal and 16 osteosarcoma tissues. The in vitro migration and invasion abilities of osteosarcoma cells were determined by a Transwell assay. A spontaneous metastatic osteosarcoma model was established to assess osteosarcoma metastasis in vivo. The N-linked glycosylated amino acids were identified by PNGase F treatment combined with Western blotting. The mechanism of TWIST1 regulating PCOLCE transcription was elucidated by luciferase, qPCR and ChIP assays. Results: PCOLCE was markedly up-regulated in human osteosarcoma tissues compared to its expression in noncancerous adjacent tissues; high PCOLCE expression in tissues correlated with a poor patient prognosis, and the knockdown of PCOLCE by shRNAs impaired the migration, invasion and lung metastasis of osteosarcoma cells. The overexpression of wild-type PCOLCE, but not its N29Q mutant, promoted migration, invasion and metastasis, indicating that the glycosylation of PCOLCE at Asn29 is necessary for its functions in osteosarcoma. TWIST1, a key transcription factor in metastasis, was also overexpressed in osteosarcoma tissues and positively correlated with either PCOLCE or its potential procollagen substrates, such as COL1A1, COL1A2, COL5A1, COL8A2 and COL10A1. Conclusion: Our findings are the first to provide evidence that PCOLCE plays a critical role in promoting the lung metastasis of osteosarcoma, and this up-regulation of PCOLCE by TWIST1 may lead to a new therapeutic strategy to treat patients with metastatic osteosarcoma.