Hypoxia‐inducible factor (HIF)‐1 and HIF‐2 are heterodimeric transcription factors that mediate the cellular response to hypoxia. Their key regulatory subunits, HIF‐1α and HIF‐2α, are induced similarly by hypoxia, but their functional roles in cancer may be distinct and isoform‐specific. SW480 colon cancer cells with stable expression of siRNA to HIF‐1α or HIF‐2α or both were established. HIF‐1α‐deficient cells displayed lower rates of proliferation and migration, but HIF‐2α‐deficient cells exhibited enhanced anchorage independent growth in a soft agar assay. Xenograft studies revealed that HIF‐1α deficiency inhibited overall tumor growth, whereas deficiency of HIF‐2α stimulated tumor growth. In human colon cancer tissues, expression of HIF‐1α and to a lesser extent, HIF‐2α, was linked to upregulation of VEGF and tumor angiogenesis. However, loss of expression of HIF‐2α but not HIF‐1α was strongly correlated with advanced tumor stage. DNA microarray analysis identified distinct sets of HIF‐1α and HIF‐2α target genes that may explain these phenotypic differences. Collectively, these findings suggest that HIF isoforms may have differing cellular functions in colon cancer. In particular, HIF‐1α promoted the growth of SW480 colon cancer cells but HIF‐2α appeared to restrain growth. Consequently, therapeutic approaches that target HIF may need to consider these isoform‐specific properties. © 2008 Wiley‐Liss, Inc.

Background: Although it has been reported that different hepatitis B virus (HBV) genotypes induce different clinical characteristics in patients with chronic liver diseases (CLD), there have been few reports that have detailed the distribution of HBV genotypes in acute forms of liver disease. Methods: HBV genotypes were determined in 61 patients who had acute forms of liver disease (45 had acute self limited hepatitis (AH) and 16 had fulminant hepatitis (FH)) and in 531 patients with CLD, including 19 patients with severe acute exacerbation of CLD. We also analysed the enhancer II, core promoter, and precore region sequences for the presence of mutations. Results: Expression of genotype B in patients with acute forms of liver disease was significantly greater than in those with CLD (39.3% v 11.7%, respectively; p,0.001). Furthermore, expression of genotype B was significantly greater in patients with FH than in those with AH (62.5% v 31.1%, respectively; p = 0.027). The precore mutation A1896 and the core promoter mutation at nt 1753 and 1754 were found more frequently in FH than in AH, and genotype B was predominant in FH regardless of the presence of these mutations. Conclusions: HBV genotype B was found more frequently in patients with acute forms of liver disease than in patients with CLD, and more frequently in patients with FH than in those with AH. These results suggest that this HBV genotype may induce more severe liver damage than other viral genotypes, at least in patients from Chiba, Japan.

The transforming growth factor-b (TGF-b)-Smad signaling pathway inhibits the growth of human epithelial cells and plays a role in tumor suppression. The Smad4 gene is mutated or deleted in 50% of pancreatic cancers. In this study, we succeeded in establishing Smad4 knockdown (S4KD) pancreatic cancer cell lines using the stable RNA interference (RNAi) method. Smad4 protein expression was reduced dramatically and TGF-b-Smad signaling was markedly inhibited in the S4KD cell lines. The S4KD and control cells were stimulated with TGF-b and analysed using a cDNA microarray that contained 3756 genes, in order to screen for target molecules downstream of TGF-b. The microarray analysis revealed that 187 S4KD genes and 155 genes in the control cells were regulated immediately upon TGF-b stimulation. Quantitative RT-PCR analysis on several of these genes produced results that corroborated the outcome of the microarray analysis. Most of the genes in the S4KD and control cells identified by the array differed, which suggests signaling pathways that differ according to Smad4 status. Of the identified genes, 246 have not been reported previously as genes that lie downstream of TGF-b. Genes that are involved in cell proliferation, adhesion, and motility were found to be regulated differentially with respect to S4KD and control cells. Cell migration induced by TGF-b was inhibited in the S4KD cells, which might be associated with a different regulation of integrin b7. The knock down of a specific gene using stable RNAi appears to be a promising tool for analysing endogenous gene function.

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