Supplementary Table 1-9. Supplementary Table 1. Clinicopathologic features of 245 HCC patients with PVTT in training cohort. Supplementary Table 2. Clinicopathologic features of 372 HCC Patients in validation cohort. Supplementary Table 3. Primers and siRNA Sequences. Supplementary Table 4. Antibodies used in the study. Supplementary Table 5. lncRNAs expressed differentially both in PVTT Compared with PT and in CSQT-2 Compared with Hep3B. Supplementary Table 6. mRNAs expressed differentially both in PVTT compared with PT and in CSQT-2 compared with Hep3B. Supplementary Table 7. LncRNA-mRNA pairs from Blat analysis. Supplementary Table 8. The Relationship between expression of ICR/ICAM1 and clinicopathologic features for 372 HCC Patients. Supplementary Table 9. Univariate and Multivariate analyses of factors associated with survival and recurrence for 372 HCC patients
ARTICLE ABSTRACT
Purpose: Portal vein tumor thrombus (PVTT) is a major complication of hepatocellular carcinoma (HCC) and is associated with poor survival. Long noncoding RNAs (lncRNA) contribute to HCC metastasis, but whether and how lncRNAs affect PVTT development remains unclear. In the present study, a novel highly expressed lncRNA (ICAM-1–related, ICR) was identified in ICAM-1+ cancer stem cells (CSC) in HCC. This lncRNA regulated CSC properties and contributed to PVTT development.Experimental Design: We used microarray and bioinformatics analyses to identify differentially expressed lncRNAs. Real-time PCR and Western blotting were used to assess gene expression in cell lines and tumors. Sphere formation assays were performed to investigate stem cell properties of tumor cells in vitro. Retrospective and prospective studies were used to investigate the relationship between ICR expression and clinical outcomes.Results: Compared with the corresponding primary tumors, PVTT expressed different lncRNAs and mRNAs, including the upregulated lncRNA ICR and ICAM-1. ICR regulated ICAM-1 expression by increasing the stability of its mRNA through RNA duplex formation, which modulated the CSC properties of ICAM-1+ HCC cells. ICR transcription in ICAM-1+ HCC cells was regulated by Nanog, and inhibition of ICR in situ significantly reduced ICAM-1 expression and ICAM-1+ HCC cells in tumors in vivo. Moreover, elevated ICR and ICAM-1 expression in tumors was correlated with PVTT development and poor clinical outcomes.Conclusions: Our study demonstrates that ICR specifically regulates CSC properties of ICAM-1+ HCC cells and that ICR contributes to PVTT development. Therefore, ICR may be a promising target for HCC therapy. Clin Cancer Res; 22(8); 2041–50. ©2015 AACR.
