(B) The number of apoptotic cells (Annexin

V+ cells) was not significantly affected in PLC/PRF/5 and HLE cells by modulating the expression of the miR-216a/217 cluster. The Annexin V+ cells decreased from 2.44% to 1.32% by overexpression of miR-216a/217 in PLC/PRF/5 cells and increased from 0.99% to 2.92% following the knockdown of miR-216a/217 in HLE cells (P>0.05). Figure S5. Expression of SMAD7 (A), PTEN (B), JAK2 (C) and CADM1 (TSLC1) (D) was shown by dot plot analysis, Barasertib in vitro by searching a HCC Gene Expression database established in our laboratory using Affymetrix Human Genome U133 plus 2.0 Arrays (Affymetrix, Santa Clara, CA, USA) comprising of HCC tumor and adjacent histologically normal liver tissue (1). Figure S6. Potential targeting region of miR-216a/217 predicted for PTEN-3′UTR (A and B) and SMAD7-3′UTR (C and D) using RNAhybrid 2.2. (A-D) The predicted target sequences and mutations generated for the 3′-UTR of PTEN and SMAD7 mRNA are shown. (E) Images to show the morphological changes observed for PLC/PRF/5-miR-216a/217 cells following transfection with wild-type plasmids containing SAMD7 (i and ii) or PTEN (iii and iv)

compared to control vectors, the morphological changes were indicative of mesenchymal-epithelial transition (MET). HIF inhibitor Figure S7. (A and B) TGF-β1 treatment induced the up-regulation of miR-216a/217 in HepG2 cells. (C and D) Addition of LY2109761, a selective TGF-β receptor type I/II dual inhibitor, inhibited TGF-β1-induced miR-216a/217 expression in HCC cells. (E) Low concentration of LY2109761 (< 1 μM) have insignificant effect on the viability of the PLC/PRF/5 cells.

Figure S8. (A) Kaplan-Meier survival analysis between HCC patients with early-recurrent and non-recurrent disease. Significant difference in disease-free survival (P<0.0001) was observed between HCC patients with early-recurrent and non-recurrent disease. (B) Immunohistochemical studies of the expression MCE of P-Akt in matched normal, early-recurrent and non-recurrent HCC liver tissue samples (20X). Of note, more than 50% of the early-recurrent HCC tissues studied by IHC exhibited elevated P-Akt staining in 25-75% of the tumor tissues examined and revealed that a significant difference in the staining of P-Akt between the early recurrent and non-recurrent HCC tissues (S8B). Figure S9. Expression of CADM1 (TSLC1) (A) and SMAD7 (B) is down-regulated in liver cancer compared with normal liver tissues (indicated by red arrows) by searching the IST Online system (http://www.medisapiens.com/ist-online-overview/). “
“Background and Aim:  Hepatocellular carcinoma (HCC) is a common human cancer worldwide. The levels of serum clusterin in HCC patients and its potential diagnostic significance is not clear. We aimed to evaluate the clinical use of serum clusterin levels as a surveillance tool for HCC with hepatitis B virus (HBV) related cirrhosis.