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Establishment of NSCLC in situ metastasis model using GFP/A549 cells

WANG Ya-wei~1,WANG Xiu-wen~1,WEI Jun-min~1,LIU Chun-xi~2(1.Department of Oncology;2.Key Laboratory of Cardiovascular Remondeling and Function Research,ChineseMinistry of Education and Chinese Ministry of Public Health,Qilu Hospital of Shandong University,Jinan 250012,Shandong,China)  
Objective: To construct the nude mouse model of human non-small cell lung cancer(NSCLC) metastasis by using GFP/A549 cells.Methods: The vector pRNAT-U6.1/Neo was transfected into A549 cells and selected the stabilizing GFP cells with G418,and then the in vivo and in vitro activity of GFP/A549 and A549 cells were compared.Nude mice were performed orthotopic transplantation of GFP/A549 cells and killed when achieved the reserved standard.Pathological and immunohistochemistry analysis were used to detect the metastasis,and KODAK IS2000MM was used to check the dissemination and the metastasis of tumor.Results: Stable clones expressing GFP were isolated,and there were no significant difference between in vivo and in vitro growth of two cell lines(P0.05).The NSCLC metastasis model was established via orthotopic transplantation.The levels of CEA and LRP of primary tumors and metastases determined by LEICA IM50 had no significant difference(P0.05).Pathological and immunohistochemistry examinations discovered brain metastases were found in 11 mice,liver metastases in 4,whereas KODAK IS2000MM showed 9 and 4,respectively.Both had no significant difference((P0.05).) Conclusion: GFP labeled A549 cells orthotopic transplantation can create NSCLC metastasis model more easily than transitional methods.KODAK IS2000MM could be used to monitor tumor metastasis and dissemination non-invasively.
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