As noted above, DFS of rectal cancer patients with overexpression of the two-protein combined panel was significantly worse than that of patients with lower expression. When patients were grouped according to LN involvement, high expression of the two-protein combined panel was associated with decreased DFS in patients with LN-positive tumors (P = 0.029; Fig 2B) but not in those with LN-negative tumors (P = 0.480; Fig 2A). By multivariate Cox analysis, expression of the two-protein combined panel in TC in the LN positive subgroup significantly correlated with DFS (HR = 3.474, 95% CI 1.054–11.451, P = 0.041) (Table 3).