ABSTRACT
ObjectiveTo detect the mRNA and protein expressions of Bmi-1 in hepatocellular carcinoma (HCC) tissue,pericarcinomatous tissue,portal vein tumor thrombus and normal liver tissue,and to investiage the significance of Bmi-1 in the genesis and progression of HCC.MethodsForty tissues of HCC were collected from the Tongji Hospital of Huazhong University of Science and Technology from January 2005 to December 2009.The mRNA and protein expressions of Bmi-1 in the HCC tissues (40 cases),pericarcinomatous tissues (40 cases),portal vein tumor thrombus ( 11 cases) and normal liver tissues ( 10 cases) were detected by immunohistochemistry,Western blot and real-time polymerase chain reaction.The relationship between the expressions of Bmi-1 and the clinicopathological factors was analyzed.Differences in each group were compared by using the Nemenyi test or Dunnet t test,and the relationship between the clinicopathological factors and the protein expression in the HCC tissues was analyzed by the chi-square test or Fisher exact probability.The survival curve was drawn by the Kaplan-Meier method,and the survival rate was analyzed by the Log-rank test.Results The median relative mRNA expressions of Bmi-1 in the normal liver tissues,pericarcinomatous tissues,HCC tissues and portal vein tumor thrombus were 0.96,2.60,7.51 and 29.95,respectively.The results of immunohistochemistry showed that the high protein expression rates of Bmi-1 in the normal liver tissues,pericarcinomatous tissues,HCC tissues and portal vein tumor thrombus were 10.0％,20.0％,67.5％ and 100.0％,respectively.The high protein expression rates of Bmi-1 in the HCC tissues and portal vein tumor thrombus were significantly higher than those in the normal liver tissues and pericarcinomatous tissues (x2 =17.25,22.77;22.04,23.95,P ＜ 0.05 ). High protein expression of Bmi-1 was also detected in 11 cases of HCC tissues with portal vein tumor thrombus.The results of western blot were consistent with those of the immunohistochemistry.The mRNA and protein expressions of Bmi-1 were correlated with Edmondson grade and portal vein metastasis ( x2 =5.572,P ＜ 0.05 ),whereas they were irrelevant to the tumor size,serum levels of α-fetoprotein and hepatitis B surface antigen ( x2 =0.000,0.019,0.663,P ＞0.05).Patients with high expression of Bmi-1 had poor prognosis.ConclusionsBmi-1 is correlated with the genesis and progression of HCC as well as the formation of portal vein tumor thrombosis.Patients with high Bmi-1 expression have poorer prognosis when compared with those with low Bmi-1 expression.
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This study examined the effect of long-term administration of low-dose FTY720 on survival of murine cardiac allograft and the possible mechanism. Murine models of abdominal heterotopic heart transplantation were established. Low-dose FTY720 (0.3 mg/kg) was administrated to the animals 4 days before the transplantation of cardiac allografts until the occurrence of rejection or the observation terminals. The animals without FTY720 treatment and those with syngeneic cardiac grafts transplanted served as controls. The mean survival time (MST) of grafts, and T lymphocyte subsets in grafts, peripheral blood and lymphoid organs were measured by histopathological examination or flow cytometry, and compared among groups. The results showed that the MST of allografts in FTY720-treated mice was more than 40 days, significantly longer than that in the untreated group (MST=8 days, P<0.01). After the long-term administration of FTY720, the proportion of CD4(+) and CD8(+) lymphocytes in peripheral blood was diminished significantly, but the proportion of CD4(+) lymphocytes was increased in mesenteric lymph nodes (MLNs) and spleen. Immunofluorescence staining revealed that the infiltration of CD4(+) and CD8(+) lymphocytes in allografts was significantly inhibited after long-term administration of low-dose FTY720. It was concluded that low-dose long-term administration of FTY720 could promote T lymphocytes in lymphatic organs and decrease their infiltration in allografts, resulting in the inhibition of rejection and the long-term survival of allografts.
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Objective To investigate the role of toll-like receptor 4 (TLR4) of endothelium or bone marrow derived cells in the acute lung injury (ALI) induced by lipopolyscccharide (LPS) in mice with reciprocal bone marrow transplantation. Method Chimeric mice were produced by reciprocal bone marrow transplantation between TLR4mut/mut and TLR4+/+ mice and divided into 4 groups: WT/WT (recipient/donor),WT/Mutant, Mutant/WT and Mutant/Mutant group. Six to eight weeks following transplantation, LPS was injected inot mice's tail vein in order to produce ALI model,and mice were sacrificed five hours later on.Samples of lung tissues were taken for the following analysis of wet/dry weight (W/D), lung permeabifity index (LPI), myeloperoxidase (MPO),levels of cytokines (TNF-α, IL-1β) and adhesion molecules (ICAM-1). Results Lung injury in the Mutant/Mutant mice was the mildest in the 4 groups. And lung injury in WT/Mutant mice was more serious than that in Mutant/WT mice. levels of MPO and ICAM-1 in WT/Mutant mice were much higher than those of Mutant/WT. In addition,the expression of ICAM-1 in WT/Mutant mice is comarable to that in WT/WT mice. Mutant/WT mice expressed higher levels of TNF-α and IL-1β than WT/Mutant mice. Conclusions Endothelial cell derived TLR4 plays ker-nel role in ALI induced by LPS via lung PMN recruitment,although bone marrow cells derived TLR4 are more im-portant for the release of cytokines.
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Objective To investigate the expression of livin in hepatocarcinoma tissues and its relationship with hepatitis B virus X protein. Methods Immunohistochemistry was used to detect the expression of livin in 30 cases of hepatic carcinoma tissues and 10 cases of normal liver tissues. Stable L02 cell hne tansfected with HBx was established. Real-time PCR and Western blot were used to observe the expression of livin gene before and after the HBx transfeetion. Results The protein expression of livin was higher in hepatocarcinoma tissues than in normal tissues, and correlated with the exist of portal vein cancer embolus (t = 2.24, P = 0.033 ; P<0.05), but it was not correlated with cancer size or classification pathologically(t = 1.688, P = 0.103 ; γ = - 0.137, P = 0.471 ; P＞0.05). Livin Mrna expression was up-regulated after the HBx gene transfection (t = 6.73, P<0.05). Conclusions Apoptosis inhibition induced by livin up-regulation may play a role in pathogenesis and development of hepatocarcinoma. HBx up-regulates livin expression at transcriptional level.
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The evolution of hepatocellular carcinoma (HCC) is a compound process which involves many kinds of genes and transductional pathways. The expression of the peptidyl-proplyl- isomerase PIN1 gene, the mutation in exon 3 of β-catenin and its correspondent abnormal expression and their roles in the hepatocellular carcinogeneisis were investigated. Among 29 pair cases of HCC and non-carcinoma tissues, the expression of PIN1 gene was detected by immunochemical staining. Mutations in exon 3 of β-catenin gene and differential expression of β-catenin gene were investigated by the methods of PCR-SSCP, direct sequencing and immunohistochemical technique as well. The results indicated: (1) 44.8% (13/29) cases of HCC presented higher level of PIN1 gene expression than non-cancerous tissues (x2 =32.63, P＜0.05), especially in cytoplasm and nucleus, while there was lower level of PIN1 expression in non-cancerous tissues; (2) 58.6% (17/29) HCC tissues showed β-catenin protein accumulation in cytoplasm and nucleus. 46.2% (6/13) HCC tissues indicated β-catenin protein accumulation with higher level of PIN1 expression, while 53.8% (7/13) HCC tissues indicated β-catenin protein accumulation with lower level or trace of PIN1 expression (x2 =0.00, P＞0.05); (3) 24.1% (7/29) of primary tumor lesions carried gene mutations in exon 3 of β-catenin, and accompanied by β-catenin protein accumulation. There was no mutation in non-cancerous tissues. All the mutation presented in tissues with low level of PIN1 expression. There was no mutation of β-catenin gene in tissues with high PIN1 expression level (x2=58.12, P＜0.05). So it was postulated that the increase of PIN1 gene expression could promote hepatocellular carcinogenesis via a way different from β- catenin gene mutation.
ABSTRACT
The evolution of hepatocellular carcinoma (HCC) is a compound process which involves many kinds of genes and transductional pathways. The expression of the peptidyl-proplyl-isomerase PIN1 gene, the mutation in exon 3 of beta-catenin and its correspondent abnormal expression and their roles in the hepatocellular carcinogeneisis were investigated. Among 29 pair cases of HCC and non-carcinoma tissues, the expression of PIN1 gene was detected by immunochemical staining. Mutations in exon 3 of beta-catenin gene and differential expression of beta-catenin gene were investigated by the methods of PCR-SSCP, direct sequencing and immunohistochemical technique as well. The results indicated: (1) 44.8% (13/29) cases of HCC presented higher level of PIN1 gene expression than non-cancerous tissues (chi2=32.63, P0.05); (3) 24.1% (7/29) of primary tumor lesions carried gene mutations in exon 3 of beta-catenin, and accompanied by beta-catenin protein accumulation. There was no mutation in non-cancerous tissues. All the mutation presented in tissues with low level of PIN1 expression. There was no mutation of beta-catenin gene in tissues with high PIN1 expression level (chi2=58.12, P<0.05). So it was postulated that the increase of PIN1 gene expression could promote hepatocellular carcinogenesis via a way different from beta-catenin gene mutation.