Anatomical trisectionectomy using anterior approach and hanging maneuver for the treatment of giant liver cancer / 中华消化外科杂志

Anterior approach refers to a method of hepatectomy which is first to resect the hepatic parenchyma and then to free the liver; hanging maneuver refers to placing a tape before the inferior vena cava for hanging the liver during hepatectomy.In October 2011,anatomical trisectionectomy was performed on a 54-year-old male patient with large hepatocellular carcinoma in the left medical lobe and right lobe with anterior approach and hanging maneuver.The diameter of the tumor was 16 cm,and was in the ⅢA/T3NOM0 stage.The indocyanine green retention at fifteen minutes was 5.4％,and the ratio of hepatic left lateral lobe volume over the standard total liver volume was 44％.The left bile duct was slightly dilated because of the compress of the tumor.The operation started with the isolation and dissection of the inflow vessels,including the right hepatic artery,the right portal vein,the middle hepatic artery,the portal vein branches of left internal lobe.The hepatic parenchyma transection was performed along the fight side of the falciform ligament.A tape was passed between the anterior surface of inferior vena cava and liver,and the liver was suspended during the transection.The left bile duct was cut at the right side of round ligament,and then the middle hepatic vein and the right hepatic vein were resected.The ligaments around the liver were dissected and the right hepatic lobe was removed.Finally,the end-toend anastomosis between the left hepatic duct and the common hepatic duct was performed.The operation lasted for 4 hours and the intra-operative blood loss was 350 mL.The patient was recovered well.At the end of 4 months after surgery,magnetic resonance cholangiopancreatography showed that the anastomosis of the bile duct was unobstructed,and there was no recurrence of tumor inside the liver.

Re-expression of RASSF1A by 5-Aza-CdR induced demethylation of the promoter region in human biliary tract carcinoma cells / 华中科技大学学报（医学）（英德文版）

Hypermethylation of the promoter region is an important mean for the transcriptional repression of a number of cancer-associated genes, and over-expression and/or increased activity of DNA methyltransferase are considered to be the main cause of promoter hypermethylation. In order to further explore the epigenetic mechanism of tumor suppressor gene RASSF1A inactivation, 5-aza-2'-deoxycytidine (5-Aza-CdR), a DNA methyltransferase inhibitor, was used to treat the human biliary tract carcinoma cell line QBC-939 at the concentration of 5 micromol/L for 24 h in this study. After the chemical intervention with 5-Aza-CdR, the methylation status in the promoter region of RASSF1A gene was detected by methylation specific PCR (MS-PCR), and the expression alteration of RASSF1A mRNA and protein were observed by RT-PCR and Western Blot respectively. Following the treatment with 5-Aza-CdR, methylation status in the promoter region of RASSF1A gene was reversed from methylation to unmethylation. A 280 bp DNA band which represented RASS1FA expression at transcriptional level and a 40 kDa (1 kDa=0.9921 ku) protein band which represented RASSF1A expression at protein level were detected by RT-PCR and Western Blot respectively in the experimental group cells and there were no corresponding bands in the control group cells. The experimental results suggest that 5-Aza-CdR can induce demethylation in the promoter region of RASSF1A. It can also reverse epigenetic transcriptional silencing caused by DNA methylation and induce the re-expression of RASSF1A in QBC-939. This study also suggest that the mechanism of RASSF1A inactivation is very closely related to the methylation of the promoter region, which may provide a new epigenetic understanding for tumor related gene inactivation and the pathogenesis of biliary tract carcinoma.

Re-expression of RASSF1A by 5-Aza-CdR Induced Demethylation of the Promoter Region in Human Biliary Tract Carcinoma Cells / 华中科技大学学报（医学）（英德文版）

Hypermethylation of the promoter region is an important mean for the transcriptional repression of a number of cancer-associated genes, and over-expression and/or increased activity of DNA methyltransferase are considered to be the main cause of promoter hypermethylation. In order to further explore the epigenetic mechanism of tumor suppressor gene RASSF1A inactivation,5-aza-2'-deoxycytidine (5-Aza-CdR), a DNA methyltransferase inhibitor, was used to treat the human biliary tract carcinoma cell line QBC-939 at the concentration of 5 μmol/L for 24 h in this study. After the chemical intervention with 5-Aza-CdR, the methylation status in the promoter region of RASSF1A gene was detected by methylation specific PCR (MS-PCR), and the expression alteration of RASSF1A mRNA and protein were observed by RT-PCR and Western Blot respectively. Following the treatment with 5-Aza-CdR, methylaiton status in the promoter region of RASSF1A gene was reversed from methylation to unmethylation. A 280 bp DNA band which represented RASS1FA expression at transcriptional level and a 40 kDa (1kDa=0.9921 ku) protein band which represented RASSF1A expression at protein level were detected by RT-PCR and Western Blot respectively in the experimental group cells and there were no corresponding bands in the control group cells. The experimental results suggest that 5-Aza-CdR can induce demethylation in the promoter region of RASSF1A. It can also reverse epigenetic transcriptional silencing caused by DNA methylation and induce the re-expression of RASSF1A in QBC-939. This study also suggest that the mechanism of RASSF1A inactivation is very closely related to the methylation of the promoter region, which may provide a new epigenetic understanding for tumor related gene inactivation and the pathogenesis of biliary tract carcinoma.

Effect of expression of PTEN and mTOR Gene Protein in the development of cholangiocarcinoma / 中国普通外科杂志

Objective To explore the expression and relationship of PTEN and mTOR in the development of cholangiocarcinoma,determine the effect of the PI3K/PTEN/AKT/mTOR signal transduction on the(development) of cholangiocarcinoma.Methods The expression of mTOR and PTEN in the cholangiocarcinoma was detected by the immunohistochemistry and RT-PCR method.Results Compared to normal tissues,the(expression) of mTOR gene in cholangiocarcinoma significantly increased,but the expression of PTEN gene(decreased).There was a negative correlation between mTOR gene and PTEN gene expression in(cholangiocarcinoma). Conclusions The expression of mTOR gene in cholangiocarcinoma was increased and the expression of PTEN was decreased.It suggested that the mTOR gene and PTEN gene could play an important role in the process of development of cholangiocarcinoma.

5-aza-2-deoxycytidine inhibits the growth of bile duct cancer cell line in vivo and in vitro / 中国普通外科杂志

Objective To study the effects of 5-aza-2-deoxycytidine (ZdCyd), a methylation inhibitor, on the growth of bile duct cancer cell line QBC939 in vivo and in vitro. MethodMTT method and flow cytometry were used to detect the growth and apoptosis of QBC939 after being treated with different dosage of ZdCyd singly or in combination with other chemotherapy drugs. Nude mouse model was used to test the effect of ZdCyd. Results5-aza-2-deoxycytidine blocks cell cycle at G1 phase increasing apoptosis rate. These effects within certain extent are dose and time dependent.5-aza-2-deoxycytidine also coordinates with other chemotherapy drugs in the anti-tumor effects. In nude mouse model the tumor occurrence rate decreased in 5-aza-2-deoxycytidine pro-treated cells of QBC939, and ZdCyd also impedes the growth of transplanted tumor. Conclusion 5-aza-2-deoxycytidine inhibits the growth of QBC939 in vivo and in vitro by inducing cell apoptosis and enhances the antitumor effect of chemotherapy drugs.

Effect of down-regulating DNMT3b expression by transfection with antisense gene on the growth of human cholangiocarcinoma cell line QBC939 / 中国普通外科杂志

Objective To study the effect of transfection with antisense DNMT3b gene eukaryotic expression plasmid on the growth of human cholangiocarcinoma cell line QBC939,and to explore the role of DNMT3b in the cholangiocarcinoma tumorigenesis.Methods The constructed antisense DNMT3b gene eukaryotic expression plasmid was transfected into QBC939 cells using liposome.The expression level of DNMT3b protein was detected by Western blot after stable transfection.The growth curves of transfected cells and un-transfected cells were observed by MTT method respectively.The cell proliferation ability was also observed by the test of colony formation in soft agar.The alterations of the cell cycle and the apoptosis rate were detected by FCM.Results Following the transfection,the protein level of DNMT3b decreased significantly;transfection with antisense DNMT3b gene eukaryotic expression plasmid did not affect the cell growth curve of QBC939,and did not decrease the cell colony formation rate(P=0.717);transfection with antisense DNMT3b gene also did not result in cell cycle alterations or induce cell apoptosis(P=0.089).Conclusions Transfection with antisense DNMT3b gene eukaryotic expression plasmid can down-regulate the expression level of DNMT3b in QBC939.It can not affect the growth and proliferation of human cholangiocarcinoma cell line QBC939,nor alter the cell cycle and induce cell apoptosis.