ABSTRACT
Gut microbiome is closely related to human health. Changes in intestinal microbial composition promote the development of tumors. Specific intestinal microorganisms and their metabolites regulate host physiological functions and tumor microenvironment, significantly affecting the anti-cancer treatment response and its adverse events. Strategies targeting gut microbiome have shown promising prospects in diagnosis and treatment of cancer.
ABSTRACT
Objective To cultivate,identify and sort bronchoalveolar stem cells(BASC)derived from normal adult mouse lung.Methods After enzymatic digestion of lung tissue with dispase and collagenase in combination,the Sca-1+ cells were isolated from the obtained pulmonary cells by magnetic cell sorting.These Sca-1+ cells were cultured in dishes coated with collagen and mouse fibroblast cell line Swiss-3T3 under a serum-free culture system for BASC,which were identified by the dual-color immunofluorescent staining clara cell specific antigen(CCA)and surfactant protein C(SP-C).Finally,these pure BASC were isolated by the flow cytometry.Results One lung of normal adult mouse could yield(1.6-1.8)?107 nucleated cells in this enzyme digestion procedure.The percentage of Sca-1+ cells we sorted from lung tissue was much higher than the unsorted [(87.3?5.9)% and(9.6?1.8)%,P
ABSTRACT
Objective To construct the eukaryotic expression vector of mouse 24p3 gene and express it in NIH3T3 fibroblast cell line and verify its biological function of delivering iron into cells. Methods 24p3cDNA was cloned by RT-PCR and the eukaryotic expression vector of mouse 24p3 gene was constructed by gene recombination technique and the recombinant plasmid was verified by restriction enzyme digestion analysis and sequenceing, then transfected into NIH3T3 cells using DOTAP transfection reagent. Then the positive clones were selected and amplified. The expression of 24p3 gene in the culture supernatant of positive clones was analyzed by ELISA and Western blotting and its biological activity of delivering iron into cells was tested by atomic absorption spectrometer. Results The eukaryotic expression vector was constructed successfully. The NIH3T3 cells that stably expressed 24p3 were screened out. 24p3/SIP24 protein could express in the culture supernatant of positive clones and deliver iron into NIH3T3 cells. Conclusion The 24p3 gene was expressed successfully in the transfected NIH3T3 cells with good biological activity, which laid the foundation for further studying its biological function and mechanism involved in the iron metabolism.