Establishment Of Metastatic Lung Cancer Animal Models And Observation By In Vivo Imaging In Mice

Objective: Establishment of metastatic lung cancer animal models, validate the metastasis-related gene expression and verify some key microRNAs that have influence on invasion and migration ability of the high metastatic human lung cancer animal model SPC-A-1sci by the molecular pathology method;To establish the fluorescent reporter gene vector, observe the development and metastasis of human lung cancer animal model in vivo, provide a reference for the application of vivo imaging technology in cancer animal model by the vivo imaging technology in small animal and based on human lung cancer animal model.Methods:Validate the differential characteristics between SPC-A-lsci (high-metastatic human lung cancer animal model) and SPC-A-1(low-metastatic human lung cancer animal model) via tail vein injection; Analyze the expression levels of several metastasis promoting genes between the high and low metastatic cell lines by RT-PCR, Western blotting, FICC, IHC and so on. The metastasis promoting genes are CD44, OPN, MMP-9, EGFR. Find some key microRNAs that have influence on invasion and migration ability of the high metastatic human lung cancer cell line SPC-A-lsci through siRNA. We established xenograft mouse models for studying human lung cancer by the fluorescent reporter gene vector, detected the imaging results of the model though the vivo imaging technology in small animal, the observed indexes included the labeling efficacy,sensitivity. Photonic Signals area, Photonic Signals strength and so on.Results:After8weeks vivo experiments,100%(12/12)of mice injected with highly metastatic cell lines developed lung metastases, while low metastatic cell lines was16.67%(2/12). Compared with the low metastatic cell lines, metastasis-associated genes CD44, OPN,MMP-9, EGFR in highly metastatic cells were significantly increased. To validate the influence of significant differences expression miRNAs on invasion and migration ability, we choosed23microRNAs to verify the influence of significant differences expression miRNAs on invasion and migration ability and find microRNA-148a and microRNA-200c can significantly inhibit the high metastatic human lung cancer cell line SPC-A-lsci invasion and migration ability. We established the method that is transfection GFP and GFP/Luc double fluorescent reporter gene vector into the cells, we labeled five cell lines, SMMC-7721, SPC-A-1, SPC-A-lsci, NCI-H460, MDA-MB-231sci, the labeling efficacy was more than90percent. The sensitivity of the vivo imaging technology in small animal is100GFP cells in vitro and1×105GFP cells in vivo or100Luc cells. We constructed the subcutaneous transplanting cancer model and orthotopic transplanting cancer model, and observed the growth and metastasis of human lung cancer animal model in vivo.Conclusions:Established the vivo imaging technology in small animal,observed the growth and metastasis of human lung cancer animal model in vivo, and the characteristics of human lung cancer animal model by the molecular pathology method, provided a reference for cancer animal model application and clinical lung study.