Objective To investigate the expression of caspase-3 in xenograft that was treated with targeted therapy with magnetic nanoparticles in nude mice. Methods QBC939 cell lines were injected into nude mice subcutaneously to establish the model of human cholangiocarcinoma xenograft. After two weeks of tumor inoculation, the animal models were divided randomly into 4 groups: group A received placebo (sodium chloride), group B were treated with magnetic nanoparticles (250 mg/kg), group C were treated with magnetic nanoparticles (150 mg /kg) combined with inner-stent, group D with magnetic nanoparticles (250 mg /kg) combined with inner-stent (the inner-stent was used to generate the magnetic targeting effect). The 21th day after treatment, expression of caspase-3 in tumor cells of each groups were measured with histochemical method and RT-PCR. Results The quantity of caspase-3 in tumor cells that were treated with magnetic nanoparticles (250 mg/kg) combined with inner-stent was the most (P＜0.05), and the quantity of caspase-3 in cells of group C was significantly more than that of the other two groups (P＜0.05). While the quantity of caspase-3 in group B was more than that of the control group(P＜0.05). Conclusion The use of magnetic nanoparticles combined with inner-stent may increase the expression of caspase-3, and the expression is dose-dependent with magnetic nanoparticles.
【Abstract】Objective To study the antitumor activity of HSVtk/CD combinative gene toward human cholangiocarcinoma in vivo. Methods Nude mouse models with transplanted subcutaneous cholangiocarcinoma were constructed and divided into 4 groups randomly, each group had 8 mice. Adenovirus solution free from suicide gene was injected in subcutaneous tumors of each mouse of control group. Adenovirus solution containing cytosine deaminase (CD), thymidine kinase (tk) and HSVtk/CD fusional gene were injected into single suicide gene either HSVtk or CD was transinfected into the tumor cells by injecting viras into subcutaneous tumor of mice of CD gene,tk gene and fused CD and tk gene group respectively. 24 hours after the injection, 5fluorocytosine (5FC) and ganciclovir (GCV) were injected introabdominally in each mouse. Growth of the tumors were monitored.Results Tumor growth of the genetransfection groups was suppressed in different degrees. Compared with the control group, the suppressing rates of the genetransfection groups were 41.2%, 55.7% and 70.0% respectively (P＜0.05). Histological examination showed good tumor growth in the control group, and tumor necrosis in the other 3 groups, particularly obvious in the group transfected with pAd(HSVtk/CD).Conclusion Combinative gene system has a b antitumor effect on cholangiocarcinoma in vivo. But it’s not powerful enough to eliminate tumor thoroughly because of insufficient “Bystander effect”.
【Abstract】ObjectiveTo study the effect of transfection with antisense DNMT3b gene eukaryotic expression vector on the expression of DNMT3b gene in human cholangiocarcinoma cell line QBC-939. MethodsThe constructed antisense DNMT3b gene eukaryotic expression vector was transfected into the human cholangiocarcinoma cell line QBC-939 by using lipofectamine transfection reagents, and positive cell clones were obtained by using G418 selection after transfection. Whether the constructed recombinant vector was transfected into QBC-939 cells successfully was confirmed by amplifying the exogenous neoR gene with PCR method. The expression of DNMT3b gene mRNA and protein were detected by semi-quantitative RT-PCR and FCM methods respectively. ResultsFollowing the transfection of antisense DNMT3b gene eukaryotic expression vector, the mRNA level of DNMT3b gene in QBC-939 cells of human cholangiocarcinoma decreased from 0.956±0.053 to 0.209±0.023, and the protein level of DNMT3b gene also decreased from (75.38±3.22)% to (29.87±3.46)%. There were very significant differences on the expression levels of DNMT3b gene between non-tranfections group and the antisense DNMT3b gene eukaryotic expression vector transfection group (P＜0.01). ConclusionTransfection with antisense DNMT3b gene eukaryotic expression vector significantly reduces the expression level of DNMT3b gene in human cholangiocarcinoma cell line QBC-939, and this study may provide a valid tool and method to investigate the function of DNMT3b gene and its role in cholangiocarcinoma.
ObjectiveTo study the relationship between the expression of sialyl Lewisx (SLeX) antigen and CD44v6 products and biological behaviors in cholangiocarcinomas. MethodsThe expression of SLeX and CD44v6 in 43 cases of cholangiocarcinoma tissue was respectively investigated by catalyzed signal amplification immunohistochemical technique.The relationship between expression of SLeX and CD44v6 and the clinicopathological factors of cholangiocarcinoma was analyzed.ResultsThe positive expression rate of SLeX and CD44v6 in cholangiocarcinoma was 67.4% and 62.8% respectively,which was significantly higher than that in control group (20.0%,P＜0.05).The high level expression of SLeX and CD44v6 were correlated with the TNM phase, differentiation degree,metastasis to lymph nodes and viscera in cholangiocarcinoma (P＜0.05). Moreover,there was a positive correlation between the SLeX and CD44v6 expression in cholangiocarcinoma (r=0.49,P＜0.001).Conclusion Expression of SLeX and CD44v6 could be helpful in predicting the biological behavior and prognosis of cholangiocarcinoma.
Nucleus plasma ratio was measured and silver-binding nucleolar organizer (AgNORs) were counted in 31 cases of cholangiocarinoma (11 cases were well-differentiated, 10 case moderately differentiated and 10 cases poorly differentiated) and in 17 cases of atypical epithelial hyperplasia related to hepatolithiasis (9 cases were simple hyperplasia, 8 cases atypical epithelial hyperplasia) by AgNORs techique and image analysis.The results showed that mucleus plasma ratio and AgNORs counts increased significantly from well-differentiated to poorly differentiated cholangiocarcinoma (Ｐ＜0.01). No statistically significant differance was shown between nucleus plasma ratio of atypical hyperplasia and well-differentiated cholangiocarinoma.The data imply that chronic proliferative cholngitis in the presence of hepatolithiasis can progress to atypical epithelial hyperplasia which may be an important precursor of cholangiocarcinoma.
In this series of 34 cases, 2 patients performed hepatic dect-jejunal anatomosis, 9 were PTCD external drainage, 8 were installation of internal drainage tubes through the PTCD, 9 were laparotories, 3 were cheemotherapeutic perfusison through artery and 3 were untreated. According to the follow-up results, the authors recommend that the internal drainage through PTCD is the better method to treat unresectable carcinoma of bile duct for proper patients.
Objective To evaluate the expression of γ-synuclein protein （SNCG） in carcinoma of bile duct andnormal bile duct tissues， and its clinical significance. Methods The expressions of SNCG were detected by SP immuno-histochemical in 60 cases of cholangiocarcinoma and 34 cases of normal bile duct tissues， and to analysis its relationship with the clinical pathological characteristics of cholangiocarcinoma. Results The positive expression rate of SNCG in carcinoma of bile duct tissues was 73.33% （44/60）， which was higher than that in normal bile duct tissues （P＜0.01）. The positive expression of SNCG in carcinoma of bile duct tissues was correlated with the depth of tumor invasion and lymph node metastasis （P＜0.01）， but not related to patients’ age， gender， and the degree of tumor differentiation （P＞0.05）. Conclusions The expression of SNCG is correlated with the cholangiocarcinoma occurrence， development， invasion， and metastasis. SNCG plays an important role in the infiltration and metastasis of carcinoma of bile duct. SNCG is expected to become a new cancer tumor marker， which can provide a basis to prognosize and to formulate the corres-ponding therapy plan.
ObjectiveTo establish a predictive model for survival and study it’s clinical value by reviewing the information of patients with hilar cholangiocarcinoma. MethodsMedical record of 196 patients with hilar cholangiocarcinoma were analyzed retrospectively. Seventeen possible clinicopathologic factors were selected. Cox model was used for univariate and multivariate analysis. Prognostic index (PI) was calculated based on the results of multivariate analysis. Patients with different PI were divided into three different risk level groups in order to compare the survival rate. Individual expected survival rate was calculated based on the median PI. Log cumulative hazards function plot was used to test Cox model proportional hazards assumption (PH assumption). ResultsThe significant prognostic factors influencing the survival rate were surgical procedure, surgical margin, and preoperative total bilirubin level (Plt;0.05). The predictive formula was PI=0.815×preoperative total bilirubin level+0.580×surgical margin－0.713×surgical procedure. According to the value of PI, all patients were divided into 3 groups, low risk group (PI≤－0.642), middle risk group (－0.642lt;PIlt;1.364), high risk group (PI≥1.364), and survival rate declined between groups and in groups with statistically significant difference (Plt;0.05). ConclusionThis model for survival can predict the prognosis of patients with hilar cholangiocarcinoma individually and help to conduct individual clinical therapy.
ObjectiveRecent advancements in the researches on cholangiocarcinoma (CC) related genes methylation in CC were reviewed and the clinical significances of aberrant DNA methylation for the diagnosis and treatment of CC were discussed. MethodsRelevant literatures about the relation between CC-related genes methylation and CC published recently were collected and reviewed. ResultsThe genesis of CC resulted from abnormal expressions of many genes. Many researches had shown that the abnormal methylation of CC-related genes had a close relation with CC. Epigenetic alteration had been acknowledged as an important mechanism contributing to early CC carcinogenesis. ConclusionsAbnormal methylation of CC-related genes is related with CC. The detection of CC-related genes methylation might provide new specific biomarkers for early noninvasive diagnosis of this disease. Using epigenetic agents such as azacytidine to modulate the activities of DNA methyltransferase and reverse the methylation status of CC-related gene might be an attractive strategy for future treatment of CC, which could be combined with conventional therapies.
ObjectiveTo explore the clinical significance of promoter hypermethylation of O6-methylguanine-DNA methyltransferase (MGMT) in cholangiocarcinoma. MethodsPromoter methylation status of MGMT gene and expression of MGMT protein were detected in cholangiocarcinoma by methylationspecific PCR and immunohistochemical staining, respectively. ResultsAberrant methylation of MGMT gene was detected in 17 patients (47.2%). Twentyone cases showed negative immunoreactivities. Of 21 patients with negative MGMT expression, 14 patients had aberrant methylation of MGMT gene. In 15 patients with positive MGMT expression, aberrant methylation of MGMT gene was only found in three cases. There was a negative correlation between promoter methylation status of MGMT gene and the expression of MGMT protein (rs=-0.816, Plt;0.05). Promoter methylation status of MGMT gene was related to depth of invasion, degree of differentiation, and TNM stage (Plt;0.05), but not to age of patient, gender, pathological type, and lymph node metastasis (Pgt;0.05). ConclusionsHypermethylation of MGMT promoter is a frequency molecular event in cholangiocarcinoma and may be involved in carcinogenesis. Methylation status of MGMT gene may be used to evaluate malignant degree of cholangiocarcinoma.