Patients infected with HCV genotype 3 (but not genotype 1) showed extensive

steatosis (Table 1). Because PTEN expression can undergo complex posttranscriptional regulation,5, 11 we assessed the PTEN protein level by immunohistochemistry. PTEN was homogeneously expressed in the Peptide 17 clinical trial cytoplasm and nuclei of all hepatocytes in uninfected control individuals (Fig. 1A and Table 1). Almost all HCV genotype 1–infected cases were also highly positive for PTEN, with slight variations among individuals (Fig. 1B and Table 1). However, in HCV genotype 3–infected patients, PTEN expression was significantly decreased in areas with fatty infiltration (Fig. 1C and Table 1), whereas in the absence of steatosis (n = 3), PTEN expression was comparable to that of healthy patients and genotype 1–infected patients. In contrast to what was previously

observed in the steatotic livers of HCV-negative obese patients,8 the intrahepatic PTEN mRNA levels of HCV genotype 3–infected patients were comparable to those of patients with genotype 1 (Fig. 1D). These data indicate that Selleck Obeticholic Acid PTEN expression is reduced in hepatocytes from patients infected with HCV genotype 3 (but not HCV genotype 1) through posttranscriptional mechanisms, and this PTEN reduction correlates with the accumulation of lipid droplets in these cells. The HCV core protein is sufficient to induce the formation of large lipid droplets in hepatocyte cell lines.14 In the absence of HCV, steatosis can be induced by the down-regulation of PTEN.8 Thus, we investigated whether the core protein of HCV genotype 3a could reduce PTEN expression in hepatocytes. Huh-7 上海皓元医药股份有限公司 and HepG2 cells were transduced with lentivectors encoding the core protein of HCV genotype 1b or 3a. PTEN protein and mRNA expression levels were then measured. The core protein of HCV genotype 3a (but not 1b) induced

a 50% decrease in the PTEN protein expression level in Huh-7 and HepG2 cells (Fig. 2A,B and Supporting Information Fig. 1). However, in agreement with data for liver specimens from HCV genotype 3–infected patients (Fig. 1), the core 3a protein did not significantly affect PTEN mRNA levels in these cells (Fig. 2C). We then investigated the potential posttranscriptional mechanisms by which core 3a could induce PTEN down-regulation in hepatocytes. HCV core 3a likely modulates PTEN expression indirectly because no physical interactions of core 3a and PTEN were detected with coimmunoprecipitation experiments (data not shown). PTEN down-regulation in core 3a–expressing cells was also unrelated to increased ubiquitination and proteosomal degradation, modifications of the redox status, and increased phosphorylation of PTEN, which can affect protein stability and expression (Supporting Information Fig. 2).11 In contrast, HCV core 3a induced a 3′-UTR–dependent blockade of PTEN mRNA translation.