ABSTRACT
Objective: To investigate the relationship between the radioresistance of pancreatic cancer cell SW1990 and the induction of the Epithelial-Mesenchymal Transition (EMT). Methods: The radio-resistant pancreatic cancer cell SW1990-R were established by using the method of radiating pancreatic cancer cell SW1990 step by step and repeatedly. Then the changes of the morphology of the cell was observed by inverted phase contrast microscope, the radioresistance of SW1990-R was detected by colony-forming assay, and the apoptosis rate of the two cell lines after radiation were measured by flow cytometry. Then invasiveness and EMT-related genes was measured by trans-well test and qRT-PCR. Finally, the model of transplanted tumor on nude mouse was used to confirm the relationship between the radioresistance of pancreatic cancer cell SW1990 and the induction of EMT. Results: Compared with SW1990, SW1990-R had a lower radiosensitivity (survival fraction in 2 Gy, SF2: 0.326 3±0.007 3 vs 0.840 8±0.001 9, P<0.05) and lower apoptosis rate[(6.12±1.27) % vs (16.87±1.73)%, P<0.05]. Meanwhile, the invasive ability of SW1990-R were significant higher than that of SW1990 cell. According to the result of both in vivo and in vitro experiment, SW1990-R had a higher expression level of Vimentin and Snail, and lower expression level of E-cadherin when compared with SW1990. Conclusion: Compared with SW1990, the radio-resistant pancreatic cancer cell SW1990-R can induce the Epithelial-mesenchymal transition.
Subject(s)
Pancreatic Neoplasms , Animals , Apoptosis , Cadherins , Cell Line, Tumor , Epithelial-Mesenchymal Transition , Mice , PancreasABSTRACT
In order to isolate the putative zona pellucida-binding proteins (ZPBPs) from boar spermatozoa, a new, simple method has been developed. The new isolation strategy made the most of the highly specific interactions between the components of the gametes. Detergent-extracted boar sperm proteins were submitted to affinity chromatography on a ZP-Sepharose column. SDS-PAGE analyses of the retained fraction under reducing conditions revealed that in addition to a component of Mr 38,000, the predominant ZPBPs contained at least three low molecular weight proteins (Mr less than 20 kDa). The isolated ZPBPs were effective in blocking sperm-oocyte binding in vitro. Using immunofluorescence microscopy, the ZPBPs were shown to be localized primarily in the sperm head, especially in the acrosomal cap region.
Subject(s)
Membrane Proteins/isolation & purification , Spermatozoa/chemistry , Zona Pellucida/metabolism , Animals , Cell Membrane/chemistry , Cell Membrane/metabolism , Cell Separation , Chromatography, Affinity , Electrophoresis, Polyacrylamide Gel , Female , Fluorescent Antibody Technique , Male , Membrane Proteins/metabolism , Sperm-Ovum Interactions , Spermatozoa/metabolism , SwineABSTRACT
A trisialoganglioside not found before in human tissue was isolated from showed this to be II3(NeuAc)3-LacCer or GT3. The ganglioside constituted 40% of the trisialogangliosides of human lung or 0.2 nmol ganglioside per g tissue.
