(D and E) Kaplan-Meier analyses of overall survival and recurrence-free survival in 100 patients with HCC following LT according to the expression levels of miR-20a. Decrease expression of miR-20a correlates with aggressive tumor features The relationships between miR-20a expression and clinicopathological features Metabolism inhibitor were analyzed based on the miR-20a real-time PCR readings. As

shown in Table 1, decrease expression of miR-20a in HCC was associated significantly with aggressive pathologic features, such as the largest tumor size (P = 0.014), multinodular HCC (P = 0.034) and micro-vascular invasion (P = 0.016). Decrease expression of miR-20a in HCC is associated with tumor recurrence and poor prognosis To further explore the clinical relevance of miR-20a, Kaplan-Meier and univariate FRAX597 chemical structure Cox proportional hazard regression analyses were performed. Kaplan-Meier analysis showed decrease miR-20a expression correlated with shorter

overall survival (P < 0.001; Figure 1D) and recurrence-free survival (P < 0.001; Figure 1E) of HCC patients following LT. Similarly, univariate analysis showed that miR-20a expression was associated with OS (P = 0.009; Table 2) and RFS (P = 0.015; Table 3). The other significant prognostic factors associated with OS and RFS in univariate analyses were also shown in Tables 2 and 3. Table 2 Univariate and multivariate Cox regression analyses of overall survival in 100 HCC patients following LT Parameter Univariate analysis Multivariable analysis HR 95% CI AZD1480 cell line P-value HR 95% CI P-value Age 0.875 0.912-1.172 0.169 – - – Gender 1.034 0.561-1.907 0.915 – - – HBV infection 0.342 0.261-0.745 0.230 – - – Cirrhosis 0.833 0.495-1.438 0.467 – - – Tumor size 1.319 1.012-1.894 0.021* 1.175 0.981-1.857

0.035* Tumor stage (III) 2.938 1.359-5.493 0.018* 2. 354 0.846-2.943 0.851 Histologic grade (G3/G1-2) 3.342 1.837-6.421 0.009* 1.773 0.732-3.101 0.082 Milan criteria (out) 1.756 1.043-3.433 0.017* 1.365 0.935-2.778 0.347 AFP >400 (ng/ml) 2.027 1.386-3.543 0.023* 1.569 1.031-4.603 0.031* Micro-vascular Florfenicol invasion 3.739 1.929-6.758 0.005* 2.671 1.756-5.545 0.009* miR-20a (low) 4.483 2.769-9.572 0.009* 4.937 2.221-9.503 0.022* Note: *statistically significant difference. Table 3 Univariate and multivariate Cox regression analyses of recurrence-free survival in 100 HCC patients following LT Parameter Univariate analysis Multivariable analysis HR 95% CI P-value HR 95% CI P-value Age 0.849 0.713-1.275 0.746 – - – Gender 1.092 0.534-2.801 0.331 – - – HBV infection 0.583 0.228-1.144 0.192 – - – Cirrhosis 0.746 0.434-1.204 0.493 – - – Tumor size 1.632 1.031-1.918 0.011* 1.253 1.123-1.792 0.014* Tumor stage (III) 1.876 1.319-2.592 0.026* 1.348 0.935-1.813 0.365 Histologic grade (G3/G1-2) 3.731 1.774-5.103 0.024* 2.931 1.526-3.858 0.