Kaplan-Meier plots of the DFS rates of AUS patients with stage III colorectal cancer in AUS cohort.

<p>Patients were stratified by risk level according to the genomic predictors, ColoGuideEx (A), MDA114 (B), Meta163 (C), and OncoDX (D) and grouped by whether they had received adjuvant chemotherapy (CTX) or not. Int, intermediate.</p

Kaplan–Meier survival plots of the DFS rates of VI patients stratified by risk level according to the five genomic predictors (A to E).

<p><i>P</i> values are based on the log-rank test. Int, intermediate.</p

Concordance of the five genomic predictors in grouping AUS patients by risk level.

<p>Correlation was quantified using Cramer's V statistics. * Inverse correlation</p

Clinical and pathological characteristics of patients with colorectal cancer.

*<p>NA, Not Available</p

Multivariate Cox proportional hazard regression analyses of DFS with clinical variables and genomic predictors.

<p>Multivariate Cox proportional hazard regression analyses of DFS with clinical variables and genomic predictors.</p

PARK7 modulates autophagic proteolysis through binding to the N-terminally arginylated form of the molecular chaperone HSPA5

<p>Macroautophagy is induced under various stresses to remove cytotoxic materials, including misfolded proteins and their aggregates. These protein cargoes are collected by specific autophagic receptors such as SQSTM1/p62 (sequestosome 1) and delivered to phagophores for lysosomal degradation. To date, little is known about how cells sense and react to diverse stresses by inducing the activity of SQSTM1. Here, we show that the peroxiredoxin-like redox sensor PARK7/DJ-1 modulates the activity of SQSTM1 and the targeting of ubiquitin (Ub)-conjugated proteins to macroautophagy under oxidative stress caused by TNFSF10/TRAIL (tumor necrosis factor [ligand] superfamily, member 10). In this mechanism, TNFSF10 induces the N-terminal arginylation (Nt-arginylation) of the endoplasmic reticulum (ER)-residing molecular chaperone HSPA5/BiP/GRP78, leading to cytosolic accumulation of Nt-arginylated HSPA5 (R-HSPA5). In parallel, TNFSF10 induces the oxidation of PARK7. Oxidized PARK7 acts as a co-chaperone-like protein that binds the ER-derived chaperone R-HSPA5, a member of the HSPA/HSP70 family. By forming a complex with PARK7 (and possibly misfolded protein cargoes), R-HSPA5 binds SQSTM1 through its Nt-Arg, facilitating self-polymerization of SQSTM1 and the targeting of SQSTM1-cargo complexes to phagophores. The 3-way interaction among PARK7, R-HSPA5, and SQSTM1 is stabilized by the Nt-Arg residue of R-HSPA5. PARK7-deficient cells are impaired in the targeting of R-HSPA5 and SQSTM1 to phagophores and the removal of Ub-conjugated cargoes. Our results suggest that PARK7 functions as a co-chaperone for R-HSPA5 to modulate autophagic removal of misfolded protein cargoes generated by oxidative stress.</p

Reactive oxygen species modulator-1 (Romo1) predicts unfavorable prognosis in colorectal cancer patients - Fig 5

<p>(A) Western blotting was performed to confirm upregulation of Romo1. (B) The cell viability of the colorectal cancer (CRC) cells was determined by MTT assay after transfection with pFlag-c1 or pFlag-c1 Romo1. Romo1 had no effect on cell viability and proliferation in CRC cells. (C) The effects of Romo1 on cell motility in CRC cells were determined by wound healing assay. Cell monolayers were scratched with a pipette tip and incubated with 5% FBS medium. Cell migration to the wound area was then monitored for 0hr, 24hr, and 48h post-wound, and the percentage of total area covered by the cells was then assessed using the NIH Image program. (D) Matrigel invasion assay was performed to determine the effects of Romo1 on the invasive ability of CRC cells. The invaded cells on the bottom chamber were stained with crystal violet and counted. Images of invasive HCT116 and DLD-1 cells are shown.</p

The cumulative overall survival rates of the whole cohort by Romo1 expression levels.

<p>P value was defined by the log rank test.</p

Reactive oxygen species modulator-1 (Romo1) predicts unfavorable prognosis in colorectal cancer patients - Fig 4

<p>(A) Western blotting was performed to confirm knockdown of Romo1. (B) The cell viability of the colorectal cancer (CRC) cells was determined by MTT assay after transfection with Romo1 siRNA, Romo1 shRNA, control siRNA, or control shRNA. Romo1 had no effect on cell viability and proliferation in CRC cells. (C) The effects of Romo1 on cell motility in CRC cells were determined by wound healing assay. Cell monolayers were scratched with a pipette tip and incubated with 5% FBS medium. Cell migration to the wound area was then monitored for 0hr, 24hr, and 48h post-wound, and the percentage of total area covered by the cells was then assessed using the NIH Image program. (D) Matrigel invasion assay was performed to determine the effects of Romo1 on the invasive ability of CRC cells. The invaded cells on the bottom chamber were stained with crystal violet and counted. Images of invasive HCT116 and DLD-1 cells are shown.</p

Survival analyses for patients who underwent curative resection.

<p>Survival analyses for patients who underwent curative resection.</p