Effect of Over-Expression of Zinc-Finger Protein (ZFX) on Self-Renewal and Drug-Resistance of Hepatocellular Carcinoma.

BACKGROUND X-chromosome-coupled zinc finger protein (ZFX) in the Zfy protein family is abundantly expressed in both embryonic and hematopoietic stem cells (HSCs). ZFX exist in various tumor cells and is correlated with proliferation and survival of tumor cells. As a malignant tumor with high invasiveness, hepatocellular carcinoma (HCC) may present resistance against chemotherapy and features of stem cells. This study aimed to explore the expression of ZFX in HCC cells, in an attempt to illustrate the role of ZFX in tumorigenesis. MATERIAL AND METHODS The expression of ZFX in tumor tissues was quantified by RT-PCR. The ZFX expression was then silenced to evaluate the stem cell-like features of HCC cells, including self-renewal, colony formation, and cell cycle, along with the sensitivity to cisplatin. Xenograft of ZFX-overexpressed HCC on nude mice was performed to evaluate the in vivo effect of ZFX on tumor growth. RESULTS Quantitative RT-PCR showed over-expression of ZFX in 51.8% of HCC tumors. The silencing of ZFX gene inhibited the self-renewal, colony formation, and proliferation ability of HCC cells (p<0.05 in all cases) via the cell cycle arrest at G0/G1 phase, in addition to the elevated sensitivity of tumor cells to cisplatin (p<0.001). Further studies showed that binding between ZFX and promoter regions of Nanog or SOX-2 regulatory factor initiate their expression in HCC cells. The xenograft experiment indicated the potentiation of tumor growth by ZFX over-expression. CONCLUSIONS ZFX is over-expressed in HCC cells, and correlates with stem cell-like features and pleiotropic characteristics.

Recent advances in the management of hepatocellular carcinoma.

Purpose: The fifth most common cancer of the gastrointestinal system is liver cancer. It is also one of the most common cancers worldwide. The available treatment options include surgery, percutaneous ablation, and liver transplantation. Some of the latest modalities for the management of hepatocellular carcinoma (HCC) are radiofrequency ablation, trans-arterial chemoembolization, radioembolization and systemic targeted agents like sorafenib. The process of choice of a particular treatment modality in HCC depends on the tumor stage, patient performance status and liver function reserve. In the recent past with progress in research, the short-term survival of HCC has improved but recurrent disease remains a fundamental problem as the pathogenesis of HCC is a multistep and complex process. The present review is focused on recent advances in the management of HCC. This review will also provide an insight on the upcoming latest modalities including the emerging role of hepatoma-derived growth factor (HDGF) overexpression in liver fibrosis and carcinogenesis.

Imbalance of tumor necrosis factor-α, interleukin-8 and interleukin-10 production evokes barrier dysfunction, severe abdominal symptoms and psychological disorders in patients with irritable bowel syndrome-associated diarrhea.

The present study aimed to explore the correlation between cytokine expression of tumor necrosis factor α (TNF­α), interleukin (IL)­8 and IL­10 with occludin production, abdominal symptoms and psychological factors in patients with irritable bowel syndrome­associated diarrhea (IBS­D). A total of 42 IBS­D patients and 20 healthy controls were included, which were recruited from QiLu Hospital in China. ELISA and immunohistochemical analysis were performed for evaluating the cytokines (TNF­α, IL­8 and IL­10) and occludin protein levels in the peripheral blood mononuclear cells (PBMCs) of all subjects. In addition, the abdominal symptoms and psychological status were assessed in IBS­D patients. Levels of TNF­α and IL­8 in the PBMCs of patients with IBS­D were significantly higher than those in the controls (P<0.001 and P=0.007, respectively), while IL­10 levels were significantly reduced in patients with IBS­D (P=0.047). Occludin production was significantly reduced in patients with IBS­D as compared with that in the controls (P<0.001). In patients with IBS­D, levels of TNF­α and IL­8 were negatively correlated with occludin levels (r=­0.34, P=0.028; r=­0.52, P<0.001, respectively). IL­10 showed a negative correlation with occludin production (r=0.05, P=0.748). Furthermore, TNF­α, IL­8 and IL­10 levels were significantly correlated with symptoms scores (r=0.74, P<0.001; r=0.55, P<0.001; r=­0.80, P<0.001, respectively) in patients with IBS­D. Within the IBS­D group, TNF­α expression was significantly increased in patients with a self­rating depression scale (SDS) score ≥50 (P=0.004) as compared with that in patients with an SDS score <50. Furthermore, IL­8 was significantly increased in IBS­D patients with a self­rating anxiety scale (SAS) or SDS score ≥50 (P=0.016, P=0.008, respectively) as compared with that in patients scoring <50. In conclusion, the results of the present study suggested that in IBS­D, an imbalance of cytokine production evoked colonic epithelial barrier dysfunction, abdominal symptoms and psychological disorders.

MicroRNA-1297 regulates hepatocellular carcinoma cell proliferation and apoptosis by targeting EZH2.

In this study, we suggested the level of miR-1297 was downreguled in the human hepatocellular carcinoma compared to the normal cells. We demonstrate ectopic expression of miR-1297 could significantly suppress hepatocellular carcinoma cells proliferation and enhance the cell apoptosis. In vitro reporter assay suggested EZH2 is a direct target gene of miR-1297. Furthermore, knockdown of EZH2 have the same effect with miR-1297 overeexpression in hepatocellular carcinoma cells. These findings provide evidence that miR-1297 plays a key role in inhibition of the hepatocellular carcinoma cells proliferation, and enhancing cell apoptosis through targeting EZH2, and strongly suggest that ex ogenous miR-1297 may have therapeutic value in treating hepatocellular carcinoma.

Efficacy and safety of anticoagulation therapy with different doses of enoxaparin for portal vein thrombosis in cirrhotic patients with hepatitis B.

BACKGROUND: Patients with cirrhosis have a high incidence of portal vein thrombosis (PVT), and optimal management of PVT in cirrhotic patients remains unclear. Currently, there is no paper on optimal doses of enoxaparin for the management of PVT with cirrhosis. AIMS: To evaluate the efficacy and safety of anticoagulation therapy with different doses of enoxaparin for PVT in cirrhotic patients with hepatitis B. MATERIALS AND METHODS: Sixty-five patients with hepatitis B-related cirrhosis and acute PVT were treated by different doses of enoxaparin. All the patients were assigned randomly to two groups: one group received enoxaparin 1 mg/kg subcutaneously every 12 h and the other group received enoxaparin 1.5 mg/kg subcutaneously every 24 h. Clinical, biochemical evaluation, Doppler ultrasound, and contrast-enhanced computed tomography were performed during the anticoagulation treatment. RESULTS: Of the 65 patients, 51 patients (78.5%) achieved complete/partial recanalization of PVT after 6 months of anticoagulation therapy. Child-Pugh scores were lower in the 51 patients who achieved complete/partial recanalization than those of the 14 nonresponders (P<0.01). No patients showed variceal bleeding during anticoagulation therapy in the two groups. The rates of nonvariceal bleeding with the use of 1.5 mg/kg every 24 h (23.5%) were higher than those with the use of 1 mg/kg every 12 h (6.4%). CONCLUSION: Anticoagulation therapy with different doses of enoxaparin for PVT in hepatitis B patients with cirrhosis is efficient and safe, and 1 mg/kg enoxaparin subcutaneously every 12 h is a better anticoagulation regimen in the treatment of PVT in cirrhotic patients.

The analysis of gamma-glutamyl transpeptidase gene in different type liver tissues.

AIM: To probe the value of gamma-glutamyl transpeptidase (GGT) messenger RNA in monitoring canceration of liver cells and for early diagnosis of hepatocellular carcinoma (HCC), by researching the types of GGT messenger RNA (GGTmRNA) in liver tissues and peripheral blood of different hepatopathy. METHODS: The three types of GGTmRNA (A, B, C) in liver tissues and peripheral blood from the patients with HCC, noncancerous hepatopathy, hepatic benign tumor, secondary carcinoma of liver, and healthy persons were detected by reverse-transcription polymerase chain reaction (RT-PCR). RESULTS: (1) In normal liver tissues, type A was predominantly found (100.00 %), type B was not found, type C was found occasionally (25.00 %). (2) The distribution of types of GGTmRNA in liver tissues with acute hepatitis, chronic hepatitis, cirrhosis, alcoholic hepatopathy was similar as in normal liver tissues (P>0.05), but type B was found in 3 of 18 patients with chronic hepatitis (16.67 %), and also in 3 of 11 patients with cirrhosis (27.27 %). (3) There was no significant difference of types of GGTmRNA between liver tissues with hepatic benign tumor, secondary carcinoma of liver and normal liver tissues (P>0.05). (4) Type B was predominant in cancerous tissues with HCC (87.5 %), the prevalence of type B in cancerous tissues was significantly higher than that in normal liver tissues (0/12) (P<0.05), but the prevalence of type A in cancerous tissues (46.88 %) was significantly lower than that in normal liver tissues (100.00 %) (P<0.05), and the prevalence of type C (6.25 %) in cancerous was the same as that in normal liver tissues (25.00 %) (P>0.05). In noncancerous tissues of livers with HCC, the main types were type A and type B, the prevalence of type A (85.71 %, 90.48 %) and type C (14.29 %, 9.52 %) in noncancerous tissues of liver with HCC was similar as that in normal liver tissues (A: 100.00 %; C: 25.00 %) (P>0.05), but the prevalence of type B (80.95 %, 76.19 %) in noncancerous tissues of livers with HCC was significantly higher than that in normal liver tissues (0/12) (P<0.05). (5) The prevalence of type B (37.5 %) in peripheral blood with HCC was higher than that in normal person (0/12) (P<0.05). In peripheral blood, type B was found in 4 of 11 cases of HCC with serum AFP negative. CONCLUSION: The shift of types of GGTmRNA from A to B in liver tissues may be closely related to the development of HCC, and the analysis of GGT gene may provide a useful tool for early diagnosis of HCC.

Amplification of screening sensitivity through selective destruction: theory and screening of a library of carbonic anhydrase inhibitors.

A new method for identifying enzyme inhibitors is to conduct their synthesis in the presence of the targeted enzyme. Good inhibitors form in larger amounts than poorer ones because the binding either speeds up synthesis (target-accelerated synthesis) or shifts the synthesis equilibrium (dynamic combinatorial libraries). Several groups have successfully demonstrated this approach with simple systems, but application to larger libraries is challenging because of the need to accurately measure the amount of each inhibitor. In this report, we dramatically simplify this analysis by adding a reaction that destroys the unbound inhibitors. This works similar to a kinetic resolution, with the best inhibitor being the last one remaining. We demonstrate this method for a static library of several sulfonamide inhibitors of carbonic anhydrase. Four sulfonamide-containing dipeptides, EtOC-Phe(sa)-Phe (4a), EtOC-Phe(sa)-Gly (4b), EtOC-Phe(sa)-Leu (4c) and EtOC-Phe(sa)-Pro (4d), were prepared and their inhibition constants measured. These inhibitors migrated to the carbonic anhydrase compartment of a two-compartment vessel. Although higher concentrations of the better inhibitors were observed in the carbonic anhydrase compartment, the concentration differences were small (1.83:1.71:1.54:1.46:1 for 4a:4b:4c:4d:5, where 5 is a noninhibiting dipeptide EtOC-Phe-Phe). Addition of a protease rapidly cleaved the weaker inhibitors (4d and 5). Intermediate inhibitor 4c was cleaved at a slower rate, and at the end of the reaction, only 4a and 4b remained. In a separate experiment, the ratio of 4a to 4b was found to increase over time to a final ratio of nearly 4:1. This is greater than the ratio of their inhibition constants (approximately 2:1). The theoretical model predicts that these ratios would increase even further as the destruction proceeds. This removal of poorer inhibitors simplifies identification of the best inhibitor in a complex mixture.

Homogeneous Catalytic Hydrosilylation of Pyridines.

The color change from yellow to violet allows the formation of the intermediate [Cp2 Ti(SiHMePh)(pyridine)] to be monitored in the reaction of PhMeSiH2 with a pyridine in the presence of the precatalyst [Cp2 TiMe2 ] [Eq. (a)]; the hydride [Cp2 TiH] is postulated to be the key intermediate in the catalytic cycle. The regioselective hydrosilylation of pyridine and also substituted pyridines can be catalyzed by the titanocene [Cp2 TiMe2 ]. R=Me, CO2 Et.