Decreased macrophage migration inhibitor factor expression up-regulats p27 in hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To observe the expression of macrophage migration inhibition factor (MIF) and p27 in hepatocellular carcinoma tissue, and to investigate the effect of MIF on the expression of p27 in hepatocellular carcinoma (HCC) cells.</p><p><b>METHODS</b>Immunohistochemistry and quantitative RT-PCR were performed to detect the expression of MIF and p27 in HCC tissues and peri-tumor tissues. Specific small interfering RNA (siRNA) targeting MIF gene was chemically synthesized and then transfected at the concentration of 50 nmol/L and 100 nmol/L into PLC cells and Hep3B cells. The mRNA levels of MIF and p27 after MIF siRNA treatment were quantified by real-time RT-PCR.</p><p><b>RESULTS</b>MIF protein and mRNA were over-expressed in the HCC tumor tissues compared to these in the peri-tumor tissues (P less than 0.01). The expression of p27 protein and mRNA was significantly lower in the HCC tumor tissues compared to these in the peri-tumor tissues (P less than 0.01). Compared to normal liver cell line L-02, HCC cell lines expressed higher level of MIF (F=61.036, P less than 0.01) and lower level of p27 (F=529.853, P less than 0.01). In MIF siRNA treated PLC and Hep3B cells, the MIF mRNA was decreased in a dose-dependent manner (F=320.1, P less than 0.01; F=201.2, P less than 0.01). The p27 mRNA was significantly up-regulated in MIF siRNA treated PLC and Hep3B cells compared to control siRNA transfected cells (F=419.4, P less than 0.01; F=459.9, P less than 0.01).</p><p><b>CONCLUSIONS</b>MIF is over-expressed in HCC tumor tissues, and the expression of p27 is repressed by MIF.</p>

Influence of liver regeneration after partial hepatectomy on the development of liver metastasis of colon cancer in rats / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the stimulated effect of liver regeneration on colon cancer cells in remnant liver in rats.</p><p><b>METHODS</b>Rat models with liver metastases or retro-peritoneal metastases of colon cancer were established: animals underwent 37% or 70% liver resection and were compared with a sham laparotomy (15, 25, 15 cases, respectively). Metastases were performed two weeks before resection. Rats were killed 3 weeks after the resection. Total body weight, liver and tumor weights were recorded. The human colon adenocarcinoma cell line Lovo was cultured in the presence of portal serum withdrawn 24 hours and 14 days after partial hepatectomy (PH). DNA synthesis was assessed by flow cytometry analysis for 5-Bromodeoxyuridine (5-BrdU) incorporation.</p><p><b>RESULTS</b>The tumor growth was accelerated in the remnant liver in 70% PH group, but the tumors in 37% PH group and retro-peritoneal site were not influenced by PH. Compared with the control group, after cultured 72 hours with portal serum withdrawn 24 h after PH, a higher 5-BrdU incorporation was found in the Lovo cell lines (P < 0.05), and it reached the peak after 120 hours of culture (P < 0.05). No difference was found between the groups when cultured with the portal serum withdrawn 14 d after PH (P > 0.05).</p><p><b>CONCLUSIONS</b>PH may accelerate the growth of residual microscopic tumor in the liver which contributes to local recurrence. It has no systemic effect and effects on the cancer cell lines in extrahepatic sites. The excision extension is related to the stimulating effects on the cancer cell line, and subtotal hepatectomy is presumably a major determinant.</p>

Interference of osteopontin expression inhibits the invasion and metastasis of human hepatocellular carcinoma cell lines / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of osteopontin (OPN) on the invasion and metastasis of human hapatocellular carcinoma (HCC).</p><p><b>METHODS</b>HCC cell lines (HCC-LM3) were transfected with the chemically synthesized small interfering RNA (siRNA). Real-time PCR and Western blot were used to quantify the mRNA and OPN protein levels. The malignant phenotypes including cellular growth, colony formation and invasion capability of the HCC cells were analyzed.</p><p><b>RESULTS</b>The OPN mRNA and proteins levels were decreased by 75% and 80% in OPN siRNA treated cells. Colony formation and migratory capability were reduced in OPN siRNA treated cells (P < 0.05).</p><p><b>CONCLUSION</b>The specific siRNA is able to reduce the OPN expression at both the mRNA and protein levels and significantly inhibits the invasiveness of HCC cells.</p>

The roles of caveolin-1 and endothelial nitric oxide synthase in the development of portal hypertension in rats with liver cirrhosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the relationship between the dynamic changes of caveolin-1 with the degrees of liver fibrosis and portal venous pressure (PVP) in the process of rat liver cirrhosis formation induced by dimethylnitrosamine (DMN); also to investigate the mechanisms of caveolin-1 in the regulation of endothelial nitric oxide synthase (eNOS).</p><p><b>METHODS</b>Liver cirrhosis was induced in rats by DMN. The degrees of liver fibrosis and PVP were measured. NOS activity was assessed by citrulline generation. Protein expressions of caveolin-1, eNOS and caveolin-1-eNOS interactions were examined by Western blot and immunoprecipitation, respectively.</p><p><b>RESULTS</b>Four weeks after DMN administration, liver fibrosis was at its peak and then decreased gradually. Immunoprecipitation and Western blot demonstrated that there was enhanced binding of caveolin-1 with eNOS in the process of rat liver cirrhosis. An increase in caveolin-1 expression was detected but the expression of eNOS was lower in cirrhotic tissues than in normal liver tissues. Caveolin-1 protein levels were positively correlated with the degrees of liver fibrosis and the levels of PVP (r=0.967, P < 0.01; r=0.922, P < 0.01, respectively), while NOS catalytic activity was negatively correlated with the degrees of liver fibrosis and levels of PVP (r= 0.973, P < 0.01; r=-0.947, P < 0.01) respectively.</p><p><b>CONCLUSIONS</b>Caveolin-1 upregulation is associated with the development of portal hypertension in liver cirrhosis. Over-expression of caveolin-1 in perisinusoidal cells may promote caveolin-1-eNOS binding and reduce the activity of eNOS.</p>

Expression of MIF, VEGF and p16 proteins and their correlation with clinicopathological features in cervical cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression of macrophage migration inhibitory factor (MIF), p16 and vascular endothclial growth factor (VEGF) proteins and their relationship with clinicopathological features in cervical cancer.</p><p><b>METHODS</b>Tissue microarray (TMA) and immunohistochemistry were used to detect the expression of MIF, p16 and VEGF proteins in specimens of 10 normal cervical epithelial tissues, 18 cervical intraepithelial neoplasia (CIN II, III) and 31 cervical squamous cell carcinomas. Western blotting was used to detect the expression of MIF, p16 and VEGF proteins in fresh samples of 3 normal cervical epithelial tissues, 3 CIN (III) and 6 cervical squamous cell carcinomas (3 Ib and 3 IIb).</p><p><b>RESULTS</b>Positive expression rates of MIF were 0, 72.2% and 93.5% in the normal, CIN and carcinoma samples, 20.0%, 33.3% and 71.0% for p16, and 10.0%, 44.4% and 74.2% for VEGF, respectively. The expression rates and levels of the three genes were significantly higher in cervical carcinomas than those in CIN. MIF expression was significantly higher in the cases with lower differentiation (17 cases, P = 0.021), and was positively correlated with VEGF expression (P = 0.0045). VEGF expression rate was significantly higher in both cases of poorly differentiated carcinomas and those with stage II b carcinoma or beyond (P = 0.004, P = 0.008). p16 expression was not found to be correlated with tumor differentiation or clinical stage. It was showed by Western blotting that the expression levels of MIF, VEGF and p16 were significantly higher in the carcinomas than those in CIN or normal tissues.</p><p><b>CONCLUSION</b>Expression of MIF, VEGF and p16 are probably involved in the process of cervical carcinogenesis. MIF expression is correlated with tumor differentiation. VEGF expression is correlated with both tumor differentiation and clinical stage.</p>

Correlations of expressions of macrophage migration inhibitor factor and cyclin D1 with tumor size and metastasis of hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore the possible relationship between the expressions of macrophage migration inhibitor factor (MIF), cyclin D1, cyclin-dependent kinase 4 (CDK4), phosphorylated-retinoblastoma susceptibility gene product Rb protein (phospho-Rb) and the development of hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>93 HCC tissues and 5 normal liver tissues were used to investigate the expressions of MIF, cyclin D1, CDK4 and phospho-Rb by tissue microarray and immunohistochemistry methods.</p><p><b>RESULTS</b>The expression rates of MIF, cyclin D1, CDK4 and phospho-Rb in the HCC tissues were 71%, 41%, 82% and 14% respectively, and in the normal liver tissues, they were 0%, 0%, 80% and 20% respectively. The expression rates of MIF and cyclin D1 were significantly different between the tumor and the normal liver tissues and the expression rates of CDK4 and phospho-Rb were not significantly different between the tumor and the normal liver tissues. The rate difference (69% versus 48%) of MIF expression between the larger tumors (> 3.5 cm) and the smaller tumors (< 3.5 cm) was of statistical significance (P < 0.01). The expression rate (62%) of cyclin D1 in the tumors with metastasis was significantly higher than the expression rate (35%) in the tumors without metastasis (P < 0.05). MIF expression was positively correlated with cyclin D1 expression in the tumor tissues (P < 0.01). CDK4 and phospho-Rb expressions were not significantly associated with the tumor sizes and metastasis status.</p><p><b>CONCLUSION</b>Our results indicate that MIF and cyclin D1 might be related to the growth and metastasis of HCC.</p>

Study on Cysteinyl Leukotriene Receptor Antagonist Depressing Acute Asthma Airway Inflammation / 实用儿科临床杂志

Objective To investigate the effects of montelukast(MK),a selective cysteinyl leukotriene receptor 1 antagonist on interleukin(IL)-5 and eotaxin expression as well as serum total immunoglobulin E(IgE) production during MK treatment of mouse acute asthma airway inflammation.Methods Sensitized Balb/c mice were challenged by ovabumin(OVA)to establish the acute asthmatic mo-(del).Twenty-five mg/kg of MK(MK group) or Saline(Saline group)were given by intravenously for 3 days.Cellular infiltration of bronchoalveolar larvage fluid(BALF) were assessed by Wright staining.Production of IL-5 and eotaxin in the lung or BALF and serum IgE were determined by enzyme linked immunosorbent assay(ELISA).The expression of IL-5 and eotaxin mRNA were measured by semi-quantified reverse transcriptase-polymerase chain reaction(RT-PCR).Plasma MK level was determined by liquid chromatography.Results After 24 h OVA challenge,the numbers of total white blood cells,neutrophils and eosinophils(EOS)of BALF were(26.0?18.9)?10~7/L,(5.92?8.09)?10~7/L and(0.74?0.88)?10~7/L in MK treatment group.They were significantly reduced compared with those in Saline group,respectively(P80%.The level of IL-5 in BALF and lung tissue were(48.52?14.45) ng/L and(27.40?9.62) ng/g protein in MK group,which significantly declined compared with that in saline group;BALF eotaxin level declined too.Serum IL-5 and total IgE level were also significantly reduced;IL-5 mRNA expression in lung significantly decreased.Eotaxin and its mRNA expression in lung were not decreased significantly.Conclusion MK(exerts) its anti-inflammatory effect mainly through the suppression of IL-5 and IgE production.