Up-regulation of PCOLCE by TWIST1 promotes metastasis in Osteosarcoma

Abstract

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.

A novel risk score model based on eight genes and a nomogram for predicting overall survival of patients with osteosarcoma

“…In recent decades, the highthroughput sequencing technologies have greatly facilitated the understanding of metastasis-related genes function by decoding the genome of cancer patients [26]. An increasing number of researchers have also concentrated on exploring the underlying pathogenesis of OS metastasis [27,28]. Moreover, building a prediction model of OS metastasis was growingly important for prognostication and clinical decision-making.…”

WTAP promotes osteosarcoma tumorigenesis by repressing HMBOX1 expression in an m6A-dependent manner

“…For the subcutaneous tumor model, 1 × 10 6 shNC, shWTAP or shHMBOX1 or shWTAP/ shHMBOX1 U2OS cells seeded into mice via subcutaneous injection. Tumor volume and tumor weight were measured to analyze tumor growth as previous described 46 . For orthotopic xenograft tumor model, shNC, shWTAP, shHMBOX1, or shWTAP/shHMBOX1 U2OS cells were labeled with a luminescent dye and GFP, and injected into the cavity of the tibia of mice.…”

Targeting the CK1α/CBX4 axis for metastasis in osteosarcoma

“…CBX4 functions by transcriptionally upregulating Runx2. To identify the key molecules in the progression of osteosarcoma, RNA-seq was recently performed using 16 osteosarcoma tissues and 4 normal tissues ( Supplementary Table 1) 15 . We found that CBX4 and CBX8, but not CBX2 or CBX6, was overexpressed in all 16 osteosarcoma tissues compared with the 4 normal tissues ( Supplementary Fig.…”