Vasculogenic mimicry(VM)is a newly-defined tumor microcirculation pattern in highly aggressive malignant tumors.We recently reported tumor growth and VM formation of gallbladder cancers through the contribution of the...Vasculogenic mimicry(VM)is a newly-defined tumor microcirculation pattern in highly aggressive malignant tumors.We recently reported tumor growth and VM formation of gallbladder cancers through the contribution of the ephrin type a receptor 2(EphA2)/focal adhesion kinase(FAK)/Paxillin signaling pathways.In this study,we further investigated the anti-VM activity of norcantharidin(NCTD)as a VM inhibitor for gallbladder cancers and the underlying mechanisms.In vivo and in vitro experiments to determine the effects of NCTD on tumor growth,host survival,VM formation of GBC-SD nude mouse xenografts,and vasculogenic-like networks,malignant phenotypes i.e.,proliferation,apoptosis,invasion and migration of GBC-SD cells.Expression of VM signaling-related markers EphA2,FAK and Paxillin in vivo and in vitro were examined by immunofluorescence,western blotting and real-time polymerase chain reaction(RT-PCR),respectively.The results showed that after treatment with NCTD,GBCSD cells were unable to form VM structures when injecting into nude mouse,growth of the xenograft was inhibited and these observations were confirmed by facts that VM formation by three-dimensional(3-D)matrix,proliferation,apoptosis,invasion,migration of GBC-SD cells were affected;and survival time of the xenograft mice was prolonged.Furthermore,expression of EphA2,FAK and Paxillin proteins/mRNAs of the xenografts was downregulated.Thus,we concluded that NCTD has potential antiVM activity against human gallbladder cancers;one of the underlying mechanisms may be via blocking the EphA2/FAK/Paxillin signaling pathway.展开更多
Objective: To investigate whether the side population cells (SP cells) exist in human gallbladder carcinoma cell line and the differences of drug resistance gene ABCG2 expression in SP cells, non-SP cells and GBC-SD c...Objective: To investigate whether the side population cells (SP cells) exist in human gallbladder carcinoma cell line and the differences of drug resistance gene ABCG2 expression in SP cells, non-SP cells and GBC-SD cell lines. Methods: Fluorescence activated cell sorter (FACS) was used to sort the SP and non-SP cells from GBC-SD cell line of gallbladder carcinoma of human being. Then, the sorting cells were cultured and detected the expression of ABCG2 gene among the SP cells, non-SP cells and GBC-SD cell lines by using reverse transcription polymerase chain reaction (RT-PCR), immunofluo-rescence chemistry, western blot and flow cytometry techniques. Results: A very small fraction cells (0.64 ± 0.08%) were iso-lated through FACS analysis which had the potency of stem cells and highly expressed ABCG2 gene (89.56 ± 3.86%). On the contrary, there were nearly no expression in non-SP cells (1.32 ± 0.49%) and lower expression in GBC-SD cell line (12.37 ± 1.61%). Conclusion: The side population cells that had the potency of stem cells existed in human gallbladder carcinoma cell line and over-expressed the drug resistance gene ABCG2. They may be play an important role in drug resistance of tumor.展开更多
文摘Vasculogenic mimicry(VM)is a newly-defined tumor microcirculation pattern in highly aggressive malignant tumors.We recently reported tumor growth and VM formation of gallbladder cancers through the contribution of the ephrin type a receptor 2(EphA2)/focal adhesion kinase(FAK)/Paxillin signaling pathways.In this study,we further investigated the anti-VM activity of norcantharidin(NCTD)as a VM inhibitor for gallbladder cancers and the underlying mechanisms.In vivo and in vitro experiments to determine the effects of NCTD on tumor growth,host survival,VM formation of GBC-SD nude mouse xenografts,and vasculogenic-like networks,malignant phenotypes i.e.,proliferation,apoptosis,invasion and migration of GBC-SD cells.Expression of VM signaling-related markers EphA2,FAK and Paxillin in vivo and in vitro were examined by immunofluorescence,western blotting and real-time polymerase chain reaction(RT-PCR),respectively.The results showed that after treatment with NCTD,GBCSD cells were unable to form VM structures when injecting into nude mouse,growth of the xenograft was inhibited and these observations were confirmed by facts that VM formation by three-dimensional(3-D)matrix,proliferation,apoptosis,invasion,migration of GBC-SD cells were affected;and survival time of the xenograft mice was prolonged.Furthermore,expression of EphA2,FAK and Paxillin proteins/mRNAs of the xenografts was downregulated.Thus,we concluded that NCTD has potential antiVM activity against human gallbladder cancers;one of the underlying mechanisms may be via blocking the EphA2/FAK/Paxillin signaling pathway.
基金Supported in part by the National Natural Science Foundation of China (No. 30471963).
文摘Objective: To investigate whether the side population cells (SP cells) exist in human gallbladder carcinoma cell line and the differences of drug resistance gene ABCG2 expression in SP cells, non-SP cells and GBC-SD cell lines. Methods: Fluorescence activated cell sorter (FACS) was used to sort the SP and non-SP cells from GBC-SD cell line of gallbladder carcinoma of human being. Then, the sorting cells were cultured and detected the expression of ABCG2 gene among the SP cells, non-SP cells and GBC-SD cell lines by using reverse transcription polymerase chain reaction (RT-PCR), immunofluo-rescence chemistry, western blot and flow cytometry techniques. Results: A very small fraction cells (0.64 ± 0.08%) were iso-lated through FACS analysis which had the potency of stem cells and highly expressed ABCG2 gene (89.56 ± 3.86%). On the contrary, there were nearly no expression in non-SP cells (1.32 ± 0.49%) and lower expression in GBC-SD cell line (12.37 ± 1.61%). Conclusion: The side population cells that had the potency of stem cells existed in human gallbladder carcinoma cell line and over-expressed the drug resistance gene ABCG2. They may be play an important role in drug resistance of tumor.
