7B,C). These findings were further confirmed using Huh7 HCC cells (Fig. 7D,E). Collectively, these results demonstrate that HMGB1-mediated caspase-1 activation is required for hypoxia-induced invasion in HCC cells. To further assess the effect of HMGB1 on HCC cell invasion, constitutively active HMGB1 was stably transfected into the Hepa1-6 cell line. HMGB1 stably expressing cells (pEGFPN1-HMGB1) displayed a significant increase in cell-invasion ability, compared with vector controls (Fig. 8A). BKM120 concentration In contrast to HMGB1 overexpression, stable knockdown of HMGB1 in Hepa1-6 cells, using short hairpin RNA (shRNA)

(Supporting Fig. 5), considerably decreased the invasiveness of Hepa1-6 cells, as evidenced by the Transwell assay (Fig. 8B). To determine whether HMGB1 participates in HCC metastasis in vivo, a murine lung metastasis model was utilized. Mice were injected via the tail vein with

luciferase-expressing tumors derived from HMGB1 shRNA and vector control clones and were monitored weekly Selleck Cisplatin for bioluminescent signals. Four weeks after injection, mice were sacrificed and their lungs were examined. Bioilluminescent signals in the lungs from the control group were much stronger than from the HMGB1 shRNA group (Fig. 8C,D). Furthermore, serum HMGB1 levels from the control group were 43.48 ±10.91 ng/mL, which were much higher than that from the HMGB1 shRNA-treated group (17.12 ± 4.56 ng/mL) (Fig. 8E). Tumor nodules were also more numerous in the control than in the shRNA group (Fig. 8F) and were confirmed with histology (data

not shown), demonstrating that HMGB1 can promote metastasis. HCC remains a leading cause of cancer-related death worldwide. This is despite the fact that a number of advances in both surgical (e.g., transplantation or resection) and ablative (e.g., transcatheter arterial chemoembolization or radiofrequency ablation) techniques have developed in the past several decades20 and is reflective of the advanced nature of disease with which many patients present as well as the lack of effective chemotherapeutic agents aimed at the treatment 上海皓元医药股份有限公司 of widely metastatic disease. Though loss of tumor-suppressor gene function, oncogene activation, direct viral effects, and angiogenesis all appear to be involved in the development of HCC,21 the lack of effective chemotherapy speaks to a gap in knowledge as to the precise molecular events and pathways involved in tumor development and progression. Therefore, further elucidating such a mechanism is an important goal in developing novel strategies to both prevent and treat HCC. Hypoxia is a hallmark of diverse human solid tumors and is associated with tumor progression.8 The extent of hypoxia in a tumor may represent an independent indicator of poor prognosis22; however, the mechanism by which hypoxia affects cancer progression is still unclear.