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Objective To investigate whether the sodium arsenite inhibiting proliferation of hepatocellar carcinoma(HCC) cells and having a relation with the expression of promyelocytic leukemia (PML) protein .Methods The immunohistochemistry , Western blot analysis ,immunofluorescence assay and quantitative PCR were used to examine the PML gene and protein expression in HCC tissue and cells .Results The immunohis to chemical staining showed that the PML protein was expressed in nucleus of well‐differentiated ,moderately differentiated and poorly differentiated HCC tissues randomly selected from 15 cases of HCC under‐going hepatic resection .Western blot analysis showed that PML protein was expressed at varying levels in all 24 HCC tissue sam‐ples ,HuH7 ,HepG2 ,Hep3B ,SMMC‐7721 and L02 cells .The immunofluorescence assay demonstrated that PML protein grains were found in the nucleis of the HuH7 ,HepG2 ,Hep3B and SMCC‐7721 cells ,in which HuH7 and Hep3B expressed more PML proteins than HepG2 and SMMC‐7721 cells .24 HCC tissue samples ,Hep3B ,HepG2 ,SMCC‐7721 and HuH7 cells all expressed PML gene .Sodium arsenite not only down‐regulated the PML protein expressed in HuH7 and primary HCC cells ,but also inhibited the HCC cell growth of HepG2 ,Hep3B ,HuH7 and SMMC‐7721 ,with the exposure time extension ,the inhibiting effect of sodium arsenite on HCC cells was enhanced .Conclusion Both HCC cell lines and tissues generally express ther PML protein and gene , PML protein may serve as the molecular basis for the direct targeting effects of arsenical agents .
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Objective To observe the PML protein expression of hepatocellar carcinoma tissue and cells lines and As 2 O3 regulate its expression .Methods Immunohistochemistry was used to examine the PML protein expression of hepatocellar carcinoma tissue . Western blot analysis were used to observe PML protein expression of hepatocellar carcinoma tissue of 12 cases ,5 hepatocellar car-cinoma cell lines ,such as HuH7 ,HepG2 ,Hep3B ,SMMC-7721 ,MHC97H .Western blot analysis was used to detected the PML pro-tein expression of these hepatocellar carcinoma cell lines after 72-96 h treated with 0 .25 μg/mL of As2 O3 .Results Immunohisch-enmical staining showed that the PML protein was expressed in both cytoplasm and nucleus ,did not well-distributed in hepatocellar carcinoma cells .There was no significant differences of PML protein expressed among differently differentiated stages of hepatocel-lar carcinoma cells .Western blot analysis found that hepatocellar carcinoma tissues of 12 cases with hepatocellar carcinoma ex-pressed PML protein ,and there was significant difference of PML protein expressed among 12 cases suffer with hepatocellar carci-noma .hepatocellar carcinoma cell lines ,such as HuH7 ,HepG2 ,Hep3B ,SMMC-7721 and MHC97H all expressed PML protein ,and there was little difference of PML protein expressed among hepatocellar carcinoma cell lines .The PML protein expression of HuH7 ,HepG2 ,Hep3B ,SMMC-7721 and MHC97H cell after 72-96 h treated with 0 .25 μg/mL of As2O3 significant decreased . Conclusion Hepatocellar carcinoma tissue and cells may express PML protein ,and As2 O3 may regulate this protein expression as well .PML protein may be the target molecule of As2 O3 treating HCC .
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Objective To evaluate the curative effect of different operative methods for duodenal diverticulum. Methods Articles of operations for duodenal diverticulum that were published between January 2000 to August 2006 were retrieved from China Journal Full-text Database, and the symptom remission rate and complication rate were underwent Meta-analysis. All data were analyzed by chi-square test. Results There was significant difference in symptom remission rate between Billroth Ⅱ subtotal gastrectomy, Roux-en-Y gastrojejunostomy and diverticulectomy, diverticulum inverting suture, sphincteroplasty of Oddi (χ~2= 21.269, 4.285, 12.914, 19.266, 3.938, P < 0.05). The difference in complication rate between Billroth Ⅱ subtotal gastrectomy, Roux-en-Y gastrojejunostomy and diverticulectomy, diverticulum inverting suture reached statistical significance (χ~2 =4.164, 9.166, 7.926,4.659, 4.858, P < 0.05). Conclusion For duodenal diverticulitis and the related complications which can not be effectively managed by internal medical treatment and with obvious symptom, Billroth Ⅱ subtotal gastrectomy and Roux-en-Y gastrojejunostumy are safe and effective.
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Objective To study the expression and cellular distribution of?-catenin in hepatocellular carcinoma and in HepG2 and SMCC-7721 cells.Methods Immunohistochemistry,immunocytochemistry techniques and Western blotting analysis were used to examine the expression and distribution of?-catenin in the cell membrane,cytoplasm and nucleus of bepatocellular carcinoma tissues,HepG2 and SMCC-7721 cells.Results Immunohistochemical staining showed that the?-catenin poorly,unevenly,even not distributed on the mem- brane of hepatocellular carcinoma cells.The expression of?-catenin on the membranes of low differentiated liver cancer cells was lower than that in the highly differentiated liver cancer cells,but it was strongly expressed in the cytoplasm of hepatocellular carcinoma cells,The ex- pression of?-catenin in the membranes of hepatocellular carcinoma cells was found to be reduced obviously in 18 out of 20 patients(90%).?-catenin was found to be poorly,unevenly,even not distributed in the membrane cells in 10 patients(50%).An obvious accumulation, but with poor-distribution,of?-catenin was observed in the cytoplasm of 17 cases(85%).Significant differences in the?-catenin expres- sion in both membrane and cytoplasm were found among differently differentiated stages of liver cancer cells.In 7 of 9 cases(77.8%)with highly differentiated liver cancer cells,?-catenin was continuously and evenly distributed in both the membrane and cytoplasm,while in 4 of 5(80%)cases with poorly differentiated liver cancer cells the expression of?-catenin in the membrane was found to be significantly poor and showed a uneven distribution in the cytoplasm.The?-catenin expression was positive in the cytoplasm of 5 of 6(83.3%)cases with moderatdy differentiated liver cancer cells.No expression of?-catenin was found in the membrane of HepG2 and SMCC-7721 cells,while it was highly expressed in both cytoplasm and nucleus.Western blotting showed that the?-catenin expressed in both cytoplasm and nucleus of all the liver cancer tissue,and also in HepG2 and SM(3C-7721 cells.Conclusions The expression of?-catenin was down-regulated in the membrane of hepatocellular carcinoma cells and HepG2 and SMCC-7721 cells,while?-catenin was strongly expressed in cytoplasm,and it was obviously accumulated in the nucleus.The distribution of?-catenin was related to the stage of differentiation of cancer cells.
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Objective To isolate hepatic stem cells from adult rats,and to examine the process and morphologic changes of isolated hepatic stem cells differentiated into hepatic cells and bile duct epithelial cells under in vitro culturing conditions.Methods AAF/PH was used to stimulate the oval cells.Selective enzymatic digestion and density gradient centrifugation were employed to isolate and purify these cells from heterogeneous liver cell population.The hepatic stem cells were induced to proliferate and differentiate into hepatic parenchymal cells by the addition of dexamethasone,DMSO and several growth factors.The morphologic changes of hepatic oval cells to that of hepatic cells and bile duct epithelial cells were examined.Immunofluorescence method was used to detect the hepatic stem cells c-kit and the expression of OV6 antigen.The albumin of hepatic stem cells and the CK19 expression were identified by RT-PCR analysis.Results The activity of freshly isolated oval cells were over 90%,with different colors and size.The diameter of oval cells was about 1/4~1/6 that of hepatocytes.Large and round hepatocytes appeared first during differentiation,and the cell morphology changed when the oval cells differentiated to hepatocytes.Freshly isolated oval cells was positive for c-kit and OV6 with immunofluorescence staining,and RT-PCR analysis indicated that the albumin and CK19 mRNA were expressed in the new isolated cells.Conclusion The oval cells could be induced to differentiate into a variety of cell lineages including hepatocytes and bile duct epithelial cells in the presence of DMSO and several growth factors.Several antigens such as c-kit,OV6,CK19 and albumin may be expressed in the freshly isolated hepatic stem cells,and the course of morphologic changes may be observed under in vitro conditions of induction and differentiation.
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The levels of cyclic nucleotides(CAMP and CGMP)in the serum and braest tissues of 60 patients with breast cancer or benign breast diseases were measured.It was found that the CAMP level was significantly lower,the CGMP significantly higher and the CAMP/ CGMP ratio was reduced in the serum and breast tissues of the patients with breast cancer.Our findings suggest that CAMP and CGMP might be factors of importance in the origin and development of breast cancer.