AIM: To compare the influence of different transplant sites in bone marrow mesenchymal stem cell (MSC)-based therapy for liver fibrosis. METHODS: MSCs isolated from Sprague Dawley (SD) rats were induced into hepatocyt...AIM: To compare the influence of different transplant sites in bone marrow mesenchymal stem cell (MSC)-based therapy for liver fibrosis. METHODS: MSCs isolated from Sprague Dawley (SD) rats were induced into hepatocyte-like cells. Liver fibrosis in SD rats was induced with carbon tetrachloride. Following hepatocyte induction in vitro, 4',6-diamidino- 2-phenylindole (DAPI)-labeled MSCs were transplanted by intravenous, intrahepatic, and intraperitoneal injection. Histopathological staining, immunohistochemistry, and biochemical analysis were used to compare the morphological and functional liver regeneration among different MSC injection modalities. The expression differences of interleukins, growth factor, extracellular matrix, matrix metalloproteinases, and tissue inhibitor of metalloproteinase were examined by real-time reverse transcription-polymerase chain reaction (RT-PCR) andenzyme linked immunosorbent assay (ELISA). RESULTS: Four days after exposure to hepatocyte differentiation medium, MSCs that did not express hepatocyte markers could express α-fetoprotein, albumin, and cytokeratin 18. The results of histopathological staining, immunohistochemistry, and biochemical analysis indicated that intravenous injection is more effective at rescuing liver failure than other injection modalities. DAPI-labeled cells were found around liver lobules in all three injection site groups, but the intravenous group had the highest number of cells. PCR and ELISA analysis indicated that interleukin-10 (IL-10) was highest in the intravenous group, whereas il1β, il6, tnfα and tgfβ, which can be regulated by IL10 and are promoters of liver fibrosis, were significantly lower than in the other groups. CONCLUSION: MSC administration is able to protect against liver fibrosis. Intravenous injection is the most favorable treatment modality through promotion of IL10 expression.展开更多
AIM:To investigate the effects of small interference RNA(siRNA) targeting of Cdx2 on human gastric cancer MGC-803 cells in vitro and in vivo.METHODS:The recombinant pSilencer 4.1-Cdx2 siRNA plasmids were constructed a...AIM:To investigate the effects of small interference RNA(siRNA) targeting of Cdx2 on human gastric cancer MGC-803 cells in vitro and in vivo.METHODS:The recombinant pSilencer 4.1-Cdx2 siRNA plasmids were constructed and transfected into gastric cancer MGC-803 cells in vitro.The stable transfectants were selected.The effects of Cdx2 siRNA on growth,proliferation,cell cycle,apoptosis,migration and invasiveness of human gastric cancer MGC-803 cells were evaluated and the expression of phosphatase and tensin homolog(PTEN),caspase-9 and caspase-3 was observed in vitro by reverse transcription polymerase chain reaction(RT-PCR) and Western blotting analysis.We also investigated the effect of Cdx2 siRNA on growth of MGC-803 cells in nude mice in vivo.RESULTS:Cdx2 siRNA led to inhibition of endogenous Cdx2 mRNA and protein expression as determined by RT-PCR and Western blotting analysis.Cdx2 siRNA significantly inhibited cell growth and proliferation,blocked entry into the S-phase of the cell cycle,induced cell apoptosis,and reduced the motility and invasion of MGC-803 cells.Cdx2 siRNA also increased PTEN expression,and activated caspase-9 and caspase-3 in MGC-803 cells in vitro.In addition,siRNA targeting of Cdx2 inhibited the growth of MGC-803 cells and promoted tumor cell apoptosis in vivo in nude mice tumor models.CONCLUSION:Cdx2 was involved in regulating progression of human gastric cancer cells MGC-803.Manipulation of Cdx2 expression may be a potential therapeutic strategy for gastric cancer.展开更多
基金Supported by Millitary Medicine and Health Foundation of China, No. 08Z030
文摘AIM: To compare the influence of different transplant sites in bone marrow mesenchymal stem cell (MSC)-based therapy for liver fibrosis. METHODS: MSCs isolated from Sprague Dawley (SD) rats were induced into hepatocyte-like cells. Liver fibrosis in SD rats was induced with carbon tetrachloride. Following hepatocyte induction in vitro, 4',6-diamidino- 2-phenylindole (DAPI)-labeled MSCs were transplanted by intravenous, intrahepatic, and intraperitoneal injection. Histopathological staining, immunohistochemistry, and biochemical analysis were used to compare the morphological and functional liver regeneration among different MSC injection modalities. The expression differences of interleukins, growth factor, extracellular matrix, matrix metalloproteinases, and tissue inhibitor of metalloproteinase were examined by real-time reverse transcription-polymerase chain reaction (RT-PCR) andenzyme linked immunosorbent assay (ELISA). RESULTS: Four days after exposure to hepatocyte differentiation medium, MSCs that did not express hepatocyte markers could express α-fetoprotein, albumin, and cytokeratin 18. The results of histopathological staining, immunohistochemistry, and biochemical analysis indicated that intravenous injection is more effective at rescuing liver failure than other injection modalities. DAPI-labeled cells were found around liver lobules in all three injection site groups, but the intravenous group had the highest number of cells. PCR and ELISA analysis indicated that interleukin-10 (IL-10) was highest in the intravenous group, whereas il1β, il6, tnfα and tgfβ, which can be regulated by IL10 and are promoters of liver fibrosis, were significantly lower than in the other groups. CONCLUSION: MSC administration is able to protect against liver fibrosis. Intravenous injection is the most favorable treatment modality through promotion of IL10 expression.
基金Supported by Grants from the Natural Science Foundation of China,No.81060201 and No.81060277the Higher School Specialized Research Foundation for the Doctoral Program of China,No.20114503110002+1 种基金the Postdoctoral Science Foundation of China,No.201003342the Natural Science Foundation of Guangxi,No.2011GXNSFA018273
文摘AIM:To investigate the effects of small interference RNA(siRNA) targeting of Cdx2 on human gastric cancer MGC-803 cells in vitro and in vivo.METHODS:The recombinant pSilencer 4.1-Cdx2 siRNA plasmids were constructed and transfected into gastric cancer MGC-803 cells in vitro.The stable transfectants were selected.The effects of Cdx2 siRNA on growth,proliferation,cell cycle,apoptosis,migration and invasiveness of human gastric cancer MGC-803 cells were evaluated and the expression of phosphatase and tensin homolog(PTEN),caspase-9 and caspase-3 was observed in vitro by reverse transcription polymerase chain reaction(RT-PCR) and Western blotting analysis.We also investigated the effect of Cdx2 siRNA on growth of MGC-803 cells in nude mice in vivo.RESULTS:Cdx2 siRNA led to inhibition of endogenous Cdx2 mRNA and protein expression as determined by RT-PCR and Western blotting analysis.Cdx2 siRNA significantly inhibited cell growth and proliferation,blocked entry into the S-phase of the cell cycle,induced cell apoptosis,and reduced the motility and invasion of MGC-803 cells.Cdx2 siRNA also increased PTEN expression,and activated caspase-9 and caspase-3 in MGC-803 cells in vitro.In addition,siRNA targeting of Cdx2 inhibited the growth of MGC-803 cells and promoted tumor cell apoptosis in vivo in nude mice tumor models.CONCLUSION:Cdx2 was involved in regulating progression of human gastric cancer cells MGC-803.Manipulation of Cdx2 expression may be a potential therapeutic strategy for gastric cancer.
