Seventeen patients (2%) were co-infected with HCV and HBV. The median maximum tumor size was 24 mm (range, 5−200) and the median number of HCC nodules was three. There were 162 patients (21%) with tumor vascular invasion. The median platelet counts BTK inhibitor was 10.6 × 104/μL (range, 2.2−65.3).

Preserved liver function as Child–Pugh class A was seen in 516 patients (67%). The average observation period was 23.3 months. During observation period, EHM were diagnosed in 71 patients. The sites of newly appeared EHM were as follows: lung in 35 patients (4.4%), bone in 25 (3.1%), lymph node in 12 (1.5%) and adrenal grand in 12 (1.5%). The cumulative incidence of EHM at 0.5, 1, 2 and 5 years was 1.6%, 4.5%, 9.2% and 22.9%, respectively. The cumulative survival after the diagnosis of EHM was as follows: 59.5% at

6 months, 24.5% at 1 year, 11.2% at 2 years and 4.5% at 5 years. Among the patients who received non-curative treatment, the incidence of EHM at 0.5, 1 and 2 years was 2.0%, 6.2% and 13.0%, respectively, in the >10 × 104 platelets group; and it was 0.6%, 2.1% and 4.2%, respectively, in the ≤10 × 104 platelets group. A significant difference between these data from the two groups (P = 0.002) was observed (Fig. 1). No correlation was observed between the site of EHM and platelet counts. The 16 parameters at the time of the initial non-curative treatment were analyzed to determine the risk factors for the occurrence of EHM by using Cox’s proportional hazard model. By univariate analysis, the following parameters were significantly associated with EHM: selleck high platelet counts (>10 × 104/μL), maximum tumor size (>30 mm), number of tumors (≥4), the presence of vascular invasion, HBV infection, HCV infection, elevated selleck chemicals DCP and Child–Pugh class A (Table 3). No significant correlation was observed between splenomegaly and EHM. On multivariate analysis for the above eight parameters exhibiting significance in the univariate analysis, number of tumors (≥4) (hazard ratio [HR] = 3.38; 95% CI = 1.94−6.16; P < 0.001), elevated DCP (HR = 2.67; 95% CI = 1.43−5.25; P = 0.001) and Child–Pugh class A (HR = 2.06;

95% CI = 1.07−4.39; P = 0.02) were the risk factors for EHM. There was a tendency toward development of EHM in patients with high platelet counts (HR = 1.73; 95% CI = 0.99−3.14; P = 0.055). IN THIS WORK, we examined the relationship between EHM and clinical parameters, including platelet counts, in two different studies. In the case–control study with newly discovered HCC patients, platelet counts in EHM positive patients were higher than those in EHM negative patients. The number of tumors and presence of vascular invasion also correlated with EHM at the time of the first treatment. In the subsequent retrospective cohort study among patients who received non-curative treatment, the risk factors for EHM were identified as elevated serum DCP, multiple tumor nodules and Child–Pugh class A.

01) and splenomegaly (40% vs 63%, P= 0.01) on ultrasound, and higher Hb (P=0.01) and platelet count (P<0.001). Patients with mild-PH had higher systemic vascular resistance (1469±335 vs 1336±423 dyne.s.cm-5, P<0.01) and lower cardiac index (2.8±0.5 vs 3.3±0.9 L/min.m2, P<0.01). Clinical markers of PH, as well as HVPG, hyperdynamic circulation and liver dysfunction worsened gradually from patients with mild-PH to patients with CSPH without varices, and from these to those with varices. After propranolol,

the HVPG decreased from 7.3±1 to 6.6±1 mmHg (P<0.01) in patients with mild-PH and from 14.7±4 to 12.2±5 mmHg (P<0.01) in those with CSPH. Such a reduction Ceritinib was much higher in patients with CSPH: -16±12% vs -8±9% (P<0.01). Patients with CSPH had higher rates of decreasing HVPG ≥10% (69% vs 36%, P<0.001), ≥20% (40 vs 12%, P<0.001) and ≥30% (14% vs 0, P=0.002). CONCLUSIONS: In compensated cirrhosis, patients with mild-PH had better liver function, lower liver stiffness, less hyperdynamic circulation and lower portal pressure reduction

with propranolol than those with CSPH. These findings support the potential utility of NSBB to prevent decompensation of cirrhosis in CSPH but not in earlier stages. Disclosures: Juan G. Abraldes – Speaking and Teaching: Gore, Janssen Rosa María Morillas – Advisory Committees or Review Panels: BRISTOL, GILEAD, Abvvie; Speaking signaling pathway and Teaching: ROCHE, JANSSEN, MSD Juan Carlos Garcia-Pagan – Grant/Research Support: GORE Jaime Bosch – Consulting: Falk, Gilead Science, Norgine, ONO-USA, Intercept pharma, Exalenz, Almirall, Conatus; Grant/Research Support: Gore The following people have nothing to disclose: Càndid Villanueva, Agustin Albil-los, Joan Genescà, Jose Luis Calleja, Carles Aracil, Rafael Bañares, Maria Poca, Beatriz Peñas, Salvador Aguustin,

Oana Pavel BACKGROUND AND AIMS: The increase of intrahepatic availability of nitric oxide induced by statins makes this drugs a potential option for the treatment of portal hypertension. This clinical trial was designed to evaluate the effects of simvastatin on the hepatic venous pressure gradient (HVPG) and on the azygos vein blood flow (AzBF) in cirrhotic patients. METHODS: A prospective, randomized, this website controlled, triple-blind trial with simvastatin and placebo was conducted, including patients with cirrhosis and portal hypertension detected by abdominal ultrasound with color Doppler flowmetry or upper digestive endoscopy. Placebo or simvastatin (40 mg) was given daily; HVPG was determined by hepatic vein catheterization and AzBF was measured by color Doppler echoendoscopy (EUS-Cd), both procedures performed at the beginning and after three months of treatment. The main endpoint was a decrease in HVPG of at least 20% from baseline or to ≤ 12 mm Hg after treatment, considered clinically relevant. The correlation between HVPG and AzBF was also tested. RESULTS: Thirty four patients were prospectively enrolled in the study and 22 of them completed the 3 month-protocol.

In SLCO1B3 polymorphism, insertion (ins)/ins, ins/wild, and wild/wild genotype was present in 2.3%, 20.5%, and 77.3% of the cases, respectively, while in 0.2%, 10.5%, and 89.4% of the controls, respectively. The allele frequency of L1 insertion was 12.5% of the cases, which was significantly greater than the controls

(5.4%, P=0.012, odds ratio 2.5 [95% CI: 1.3-4.9]). The c.1738C>T mutation in SLCO1B1 was not observed in both cases and Opaganib in vitro controls. [Conclusions] The genotype of L1 retrotransposon insertion in SLCO1B3 was observed more frequently in Japanese patients with drug-induced cholestasis than controls. As L1 insertion potentially impairs the function of OATP1B3, the individuals with this polymorphism might be predisposed to acquired cholestasis. http://www.selleckchem.com/products/ABT-263.html Disclosures: The following people have nothing to disclose: Tatehiro Kagawa, Kazuya Anzai, Kota Tsuruya, Yoshitaka Arase, Shunji Hirose, Koichi Shiraishi, Tetsuya Mine Aim: The performance of single and repeated brush cytology in detecting dysplasia or cholangiocarcinoma (CCA) in patients with primary sclerosing cholangitis (PSC) prior to liver transplantation, and patients’ survival during follow-up was compared to the histopathology of the explanted liver. Methods: All consecutive PSC patients undergoing liver transplantation in Sweden between 1999 and 2013 were evaluated (n=255). Patients

were categorized using histopathology of the explanted liver to determine the presence

of CCA or dysplasia. Sensitivity, specificity, see more and other measures of test performance were calculated for single and repeated brush cytology, with or without fluorescence in situ hybridization (FISH). Survival after liver transplantation was analyzed using Kaplan-Meier estimate. Results: Brush cytology was done before liver transplantation in 117 of the 225 patients, of whom 65 patients were brushed more than once. The sensitivity and specificity of brush cytology for diagnosing dysplasia or CCA increased from 50% and 81% respectively in patients with one sampling, to 100% and 83% respectively in patients where repeated examinations were performed (table 1). When considering only the subgroup where FISH was also done in addition to brush cytology (n=64), the presence of aneuploidy increased the sensitivity of brush cytology in this subgroup from 83% to 95%, while the finding of only diploid cells increased specificity from 90% to 95%. Survival after liver transplantation was significantly lower in the group with pre-transplantation undiagnosed CCA in the explanted liver p<0.001). Conclusion: In PSC patients, the utilization of repeated brush cytology or the combination with FISH results in increased sensitivity and specificity for the detection of dysplasia or CCA.

8 Some of these pathways may also attribute to the hepatoprotection of IL-22 in alcoholic liver injury. Moreover, Yang et al. recently reported that IL-22 treatment ameliorates obesity-associated

fatty liver by down-regulating several lipogenesis- and triglyceride synthesis-related genes.12 However, we found that IL-22 treatment significantly down-regulates expression of FATP, but not other fat metabolism–associated genes (Fig. 7 and Supporting Information Fig. 3). The discrepancy between these studies may be due to the different models employed. Yang et al. used mice fed a high-fat diet for 6 months that had severe hepatic steatosis,12 whereas we used mice treated with chronic-binge feeding only for 10 days that had mild selleck inhibitor steatosis. In our model, down-regulation of FATP likely contributes to the protective effect of IL-22 on ethanol-induced fatty liver, as inactivation of FATP has been shown to ameliorate high fat diet-induced fatty liver.33 In addition, we have demonstrated that IL-22 treatment elevates expression of MT I/II (Fig. 7), two antioxidant genes that play an important role in protecting against alcoholic liver injury,27

suggesting that induction of MT I/II may contribute to IL-22 hepatoprotection against ethanol-induced hepatocellular damage. Similar to IL-22, IL-6 also activates STAT3 in hepatocytes and protects against ethanol-induced liver selleck compound injury.34 However, treatment with IL-6 may generate many side effects, such as fever and inflammation, among others,35 which is due to the ubiquitous expression of IL-6 receptors and its gp130 signal chain in a wide variety of cell types, and thereby limits its clinical application for treating patients. In contrast, IL-22 selleck screening library may have better therapeutic potential in combination with current therapy of corticosteroids or TNF-α inhibitors

in treating alcoholic hepatitis (see discussion below). Corticosteroids are widely used and TNF-α inhibitors have been tested in treating alcoholic hepatitis, but the results have been controversial.2, 4-7 This is likely because treatments with these two drugs have anti-inflammatory effects, which are beneficial for alcoholic hepatitis, but can also inhibit liver regeneration36, 37 and increase the rate of bacterial infection.4-6 The latter two events are potentially fatal to patients with severe alcoholic hepatitis and are probably responsible for the poor outcomes associated with these treatments.4-6 Findings from this study and previous studies suggest that treatment with IL-22 in combination with corticosteroids or TNF-α inhibitors may have many beneficial effects in treating alcoholic hepatitis.

“
“van Rooyen DM, Larter CZ, Selumetinib manufacturer Haigh WG, Yeh MW, Ioannou G, Kuver R, et al. Hepatic free cholesterol accumulates in obese, diabetic mice and causes nonalcoholic steatohepatitis. Gastroenterology 2011;141:1393-1403. (Reprinted with permission.) BACKGROUND & AIMS: Type 2 diabetes and nonalcoholic steatohepatitis

(NASH) are associated with insulin resistance and disordered cholesterol homeostasis. We investigated the basis for hepatic cholesterol accumulation with insulin resistance and its relevance to the pathogenesis of NASH. METHODS: Alms1 mutant (foz/foz) and wild-type NOD.B10 mice were fed high-fat diets that contained varying percentages of cholesterol; hepatic lipid pools and pathways Gemcitabine cell line of cholesterol turnover were determined. Hepatocytes were exposed to insulin concentrations that circulate in diabetic foz/foz mice. RESULTS: Hepatic cholesterol accumulation was attributed to up-regulation of low-density lipoprotein receptor via activation of sterol regulatory element binding

protein 2 (SREBP-2), reduced biotransformation to bile acids, and suppression of canalicular pathways for cholesterol and bile acid excretion in bile. Exposing primary hepatocytes to concentrations of insulin that circulate in diabetic Alms1 mice replicated the increases in SREBP-2 and low-density lipoprotein receptor and suppression of bile salt export pump. Removing cholesterol from diet prevented hepatic accumulation of free cholesterol and NASH; increasing dietary cholesterol levels exacerbated hepatic accumulation of free cholesterol, hepatocyte injury or apoptosis, macrophage recruitment, and liver fibrosis. CONCLUSIONS: In obese, diabetic mice, hyperinsulinemia alters nuclear transcriptional regulators of cholesterol homeostasis, leading to hepatic accumulation of free cholesterol; the resulting cytotoxicity mediates transition of steatosis

to NASH. Obesity in the United States and other developed countries is increasing at an alarming rate.1, 2 Among the myriad health complications associated with obesity (including diabetes and cardiovascular risk) is nonalcoholic fatty liver disease (NAFLD). NAFLD is a spectrum of liver diseases ranging from simple selleck chemical steatosis, to active inflammation (nonalcoholic steatohepatitis [NASH]), to advanced fibrosis and cirrhosis,3 to hepatocellular carcinoma.4 Risk factors for primary NAFLD (i.e., not secondary to other proximate causes) are analogous to those of the metabolic syndrome (e.g., obesity, type II diabetes, and dyslipidemia).5 The prevalence of simple steatosis in individuals at risk for NAFLD can be very high; for example, the prevalence in the severely obese (body mass index >35) has been reported to be 90%.6 In contrast, the prevalence of NASH is much lower in this population (∼40%).6 These factors emphasize that the risk for developing the more severe stages of NAFLD (i.e.

CD40-CD40L interactions are a key event in T-cell-dependent humoral immune responses [30]. The

results from studies on the significance of these interactions for the differentiation of memory B cells into ASC, however, are in conflict. Several reports suggest that CD40 signalling is important for the terminal differentiation of B cells and for antibody secretion [31–34]. Other reports show that CD40 signalling prevents the terminal differentiation of B cells [35–39]. Our results indicate that the re-stimulation of FVIII-specific memory B cells and their subsequent differentiation into anti-FVIII ASC requires CD40-CD40L interaction. The blockade of these interactions prevented the formation of anti-FVIII ASC Ixazomib in vitro in vitro and reduced it significantly in vivo [17]. We believe that the blockade of CD40-CD40L interactions in our system downregulates T-cell activation and, more importantly, blocks the interaction between activated T cells and memory B cells. Based on the successful use of high-dose FVIII for the induction of immune tolerance in patients with haemophilia A [1], we speculated on the issue of whether the re-stimulation of FVIII-specific memory B cells was affected in any significant manner by high concentrations of FVIII. Our results demonstrate that concentrations of FVIII AZD9668 clinical trial that are below the physiological plasma concentration

of 100 ng mL−1 (1 U mL−1) re-stimulate FVIII-specific memory B cells and induce their differentiation into ASC in vitro, whereas concentrations that are above the physiological plasma concentration inhibit this process. These results support the idea that the inhibition or eradication of FVIII-specific memory B cells might be an early event in the downregulation of established click here anti-FVIII antibody responses in patients. The eradication of memory B cells would prevent their differentiation into ASC and, moreover, may lead to a deficiency of effective antigen-presenting cells

required for the re-stimulation of FVIII-specific T cells. The induction of regulatory T cells rather than effector T cells could be the consequence of this deficiency. Currently, it is not clear, however, whether high-dose FVIII ITI therapy in patients would lead to local FVIII concentrations that are comparable with the concentrations that we used in our in vitro experiments. Further studies are necessary to investigate this hypothesis. Toll-like receptors recognize invading pathogens such as viruses and bacteria and serve as an important link between innate and adaptive immunity [40,41]. Given the importance of TLR for the regulation of adaptive immune responses, we speculated as to how the triggering of TLR would influence the regulation of FVIII-specific memory B cells.

9 Thus, it has been demonstrated that PTTG1 accumulation inhibits mitosis progression and chromosome segregation, but does not directly affect cytokinesis, resulting in aneuploidy.35 It has been shown that HBx can transform cultured cells21 and induce liver cancer in transgenic mice.36 Genetic instability is frequently accompanied with the acquisition of transformation ability and malignant progression of tumors. Moreover, recent reports have shown that HBx expression induces chromosomal aberrations such as chromosome rearrangements and micronuclei formation.37 selleck kinase inhibitor Furthermore,

HBx promotes multipolar spindle formation and chromosomal missegregation during mitosis, and increases multinucleated cells.18 Interestingly, it has been determined that HBx binds to BubR1, a component of the mitotic checkpoint complex, and attenuates the association between BubR1 and CDC20, an activator of the anaphase-promoting complex/cyclosome, resulting in chromosomal instability.38 Our results

demonstrate that HBx induces the accumulation of PTTG1 in interphase cells. Further experiments are necessary to study the effects of HBx on PTTG1 functions during mitotic events. In conclusion, we propose that HBx promotes alterations of PTTG1 expression levels, which may improve our understanding of the molecular mechanisms of HBV-related pathogenesis of progressive liver disease leading to cirrhosis and HCC development. We thank Drs. O. M. Andrisani, H. Cho, E. Lara-Pezzi, M. Levrero, S. Murakami, K. I. Nakayama, B. L. Slagle, and J. R.

Wands for providing critical reagents and R. López-Rodríguez for statistical Navitoclax cost analysis. Additional Supporting Information may be found in the online version of this article. “
“Recently, the management of chronic hepatitis C virus (HCV) has been greatly advanced with introduction of direct-acting antiviral agents (DAAs) in clinical setting. In Japan, the first DAA, telaprevir (TVR), was approved for patients with chronic hepatitis C in 2011. Along with this, the Japan Society of Hepatology (JSH) produced the first clinical practice guideline for the management of HCV infection, “Guidelines for the Management of Hepatitis C Virus Infection” in May 2012 (English version, 2013[1]). It is our great pleasure that these Guidelines selleck chemicals llc were welcomed and utilized by physicians and other health care providers in daily clinical practices in Japan. Meanwhile, in September 2013, a second-generation DAA, simeprevir (SMV), was approved for use in Japan. According to Phase III trials in Japan and overseas, SMV has a robust therapeutic effect with better safety profiles compared to TVR. As a result, we have decided to update the clinical guidelines for HCV with launch of this new DAA. SMV has now been approved for use in patients with chronic hepatitis C with genotype 1 and high viral load, and therefore these current Guidelines are updated for patients in this group.

pylori infection is the initiation of an inflammatory response in which cytokines are the main mediators. Genetic polymorphisms can either accentuate or attenuate the host’s response to inflammation, thus affecting the interaction with environmental factors and H. pylori, the pattern and severity of gastric inflammation and the clinical outcome. Interleukin (IL)-1β is a potent proinflammatory cytokine and is involved in the host’s response to many antigenic challenges. El Omar et al. studied the correlation of these IL-1β genotypes with hypochlorhydria LY294002 manufacturer and gastric atrophy in a Caucasian population of gastric cancer relatives. Genetic polymorphisms in the

IL-1 gene cluster significantly increased the risk of precancerous gastric lesions.27 There is a strong association between genetic polymorphisms

in the IL-1β gene cluster27–30 in tumor necrosis factor-α, IL-1031 and in the IL-8 promoter,32,33 and the risk of gastric cancer. Interestingly a meta-analysis of the role of IL-1β and IL-1 receptor antagonist gene polymorphisms in gastric cancer risk showed an association in Caucasians, but not in Asians.34 Apart from genetic polymorphisms in proinflammatory genes, differences in tumor suppressor genes may be important. RUNX3, a member of the human runt-related transcription factor family, is a possible tumor suppressor gene for gastric cancer.35 RUNX3 expression is Dabrafenib solubility dmso frequently suppressed by promoter hypermethylation in gastric cancer cell lines and tissues. A recent study showed that persistent H. pylori infection could induce RUNX3 methylation, with the subsequent loss of RUNX3 expression potentially affecting gastric carcinogenesis.36 Another recent study attempted to identify RUNX3 target genes that promote cell–cell contact. Tumorigenic RUNX3-negative gastric epithelial cells attach weakly to each other, compared with non-tumorigenic, RUNX3-positive cells. It was found that the promoter activity of the gene that encoded the tight junction protein claudin-1 was upregulated via the binding of RUNX3 to the RUNX

consensus sites. The tumorigenicity of gastric epithelial cells from RUNX3-negative mice was significantly reduced by restoration of claudin-1 expression, whereas knockdown of claudin-1 increased the tumorigenicity of human gastric click here cancer cells. It was concluded that the tight junction protein claudin-1 was a direct transcriptional target of RUNX3.37 All these serve to highlight the complex interactions between host and bacterial factors. Environmental factors that have been implicated include salt-preserved food and dietary nitrite, smoking and even metabolic disturbances such as hyperglycemia and hyperlipidemia. In an ecological study, the respective importance of high salt and nitrate intake for gastric cancer mortality was assessed at the population level in 24 countries.38 There was a significant correlation between gastric cancer mortality and sodium as well as nitrate in both men and women.

pylori infection is the initiation of an inflammatory response in which cytokines are the main mediators. Genetic polymorphisms can either accentuate or attenuate the host’s response to inflammation, thus affecting the interaction with environmental factors and H. pylori, the pattern and severity of gastric inflammation and the clinical outcome. Interleukin (IL)-1β is a potent proinflammatory cytokine and is involved in the host’s response to many antigenic challenges. El Omar et al. studied the correlation of these IL-1β genotypes with hypochlorhydria Selleckchem PF-2341066 and gastric atrophy in a Caucasian population of gastric cancer relatives. Genetic polymorphisms in the

IL-1 gene cluster significantly increased the risk of precancerous gastric lesions.27 There is a strong association between genetic polymorphisms

in the IL-1β gene cluster27–30 in tumor necrosis factor-α, IL-1031 and in the IL-8 promoter,32,33 and the risk of gastric cancer. Interestingly a meta-analysis of the role of IL-1β and IL-1 receptor antagonist gene polymorphisms in gastric cancer risk showed an association in Caucasians, but not in Asians.34 Apart from genetic polymorphisms in proinflammatory genes, differences in tumor suppressor genes may be important. RUNX3, a member of the human runt-related transcription factor family, is a possible tumor suppressor gene for gastric cancer.35 RUNX3 expression is ZD1839 solubility dmso frequently suppressed by promoter hypermethylation in gastric cancer cell lines and tissues. A recent study showed that persistent H. pylori infection could induce RUNX3 methylation, with the subsequent loss of RUNX3 expression potentially affecting gastric carcinogenesis.36 Another recent study attempted to identify RUNX3 target genes that promote cell–cell contact. Tumorigenic RUNX3-negative gastric epithelial cells attach weakly to each other, compared with non-tumorigenic, RUNX3-positive cells. It was found that the promoter activity of the gene that encoded the tight junction protein claudin-1 was upregulated via the binding of RUNX3 to the RUNX

consensus sites. The tumorigenicity of gastric epithelial cells from RUNX3-negative mice was significantly reduced by restoration of claudin-1 expression, whereas knockdown of claudin-1 increased the tumorigenicity of human gastric find more cancer cells. It was concluded that the tight junction protein claudin-1 was a direct transcriptional target of RUNX3.37 All these serve to highlight the complex interactions between host and bacterial factors. Environmental factors that have been implicated include salt-preserved food and dietary nitrite, smoking and even metabolic disturbances such as hyperglycemia and hyperlipidemia. In an ecological study, the respective importance of high salt and nitrate intake for gastric cancer mortality was assessed at the population level in 24 countries.38 There was a significant correlation between gastric cancer mortality and sodium as well as nitrate in both men and women.

2A), suggesting that AIB1 may regulate apoptosis. To determine the role of AIB1 in drug resistance of CCA cells, some common chemotherapeutic drugs including tamoxifen, cisplatin, mitomycin C, and 5-FU were used to treat CCA cells. As shown in Fig. 3 and Supporting Fig. 3, down-regulation of AIB1 enhanced the sensitivity of QBC939 and SK-ChA-1 cells to these drugs. In contrast, HCCC9810 cells were more resistant to these drugs after up-regulation of AIB1 expression. Furthermore, protein lysates obtained from cells exposed to different

doses of cisplatin were studied with western blot. The results showed that AIB1 knockdown enhanced cisplatin-induced apoptosis of QBC939 and SK-ChA-1 cells, as demonstrated by increased degradation of poly ADP-ribose polymerase (PARP) (Fig. 4A, upper and middle panels). Conversely, overexpression of AIB1 suppressed cisplatin-induced IWR-1 concentration apoptosis of HCCC9810 cells (Fig. Dabrafenib clinical trial 4A, lower panel). To investigate the role of AIB1 in CCA growth and drug resistance in vivo, we compared the growth of AIB1-knockdown and control QBC939 xenograft tumors in nude mice with or without cisplatin treatment. As shown in Fig. 4B and Supporting Fig. 4, AIB1-knockdown QBC939 tumors were much smaller than control tumors in the absence of cisplatin, demonstrating that down-regulation of AIB1 inhibits CCA growth. In the presence of cisplatin, the growth of AIB1-knockdown QBC939 tumors was

further inhibited, whereas the growth of control tumors was not affected, suggesting that down-regulation of AIB1 enhances the sensitivity of CCA to cisplatin. Consistently, PCNA expression was significantly

decreased in three representative AIB1-knockdown tumors, and knockdown of AIB1 significantly learn more enhanced the apoptosis of cells in tumors treated with cisplatin as demonstrated by increased degradation of PARP (Fig. 4C). To determine the mechanism by which knockdown of AIB1 sensitizes CCA cells to chemotherapeutic drug-induced apoptosis, the expression of several antiapoptosis genes was assessed in control and AIB1-knockdown CCA cells. The results revealed that down-regulation of AIB1 suppressed the messenger RNA (mRNA) level of Bcl-2 but not cFLIPL, Bcl-XL, or Mcl-1 in QBC939 cells (Fig. 5A). Furthermore, the effect of AIB1 on Bcl-2 protein expression was investigated by western blotting. As shown in Fig. 5B, AIB1 knockdown inhibited the expression of Bcl-2 protein in both QBC939 and SK-ChA-1 cells, whereas overexpression of AIB1 enhanced the Bcl-2 protein expression in HCCC9810 cells. In addition, down-regulation of AIB1 suppressed Akt activation in QBC939 and SK-ChA-1 cells, whereas overexpression of AIB1 promoted activation of Akt in HCCC9810 cells (Fig. 5B). Because it has been reported that Akt can up-regulate Bcl-2 expression,9 we examined whether inhibition of Akt activation by LY294002 can down-regulate Bcl-2 expression in CCA cells. As shown in Fig.