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山东大学学报(医学版)

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利用GFP/A549建立人NSCLC裸鼠原位种植转移模型

王亚伟1,刘春喜2,王秀问1   

  1. 山东大学齐鲁医院 1.肿瘤内科; 2. 心血管重构和功能研究教育部重点实验室, 山东 济南 250012
  • 收稿日期:2005-12-16 修回日期:1900-01-01 出版日期:2006-07-24 发布日期:2006-07-24
  • 通讯作者: 王亚伟

WANG Ya-wei1,LIU Chun-xi2,WaNG Xiu-wen1,WANG Xiu-mei1   

  • Received:2005-12-16 Revised:1900-01-01 Online:2006-07-24 Published:2006-07-24
  • Contact: WANG Ya-wei

摘要: 目的:利用人肺腺癌细胞株GFP/A549建立非小细胞肺癌(nonsmall cell lung cancer, NSCLC)转移模型。方法:将表达GFP的质粒pRNATU6.1/Neo转染人肺腺癌细胞A549,G418筛选获得稳定表达GFP细胞,对比转染前后细胞的生长活性和成瘤性。将转染后细胞原位种植裸鼠预定标准处死。HE染色和免疫组化检测转移灶的位置和数目。利用KODAK IS2000MM系统检测肿瘤播散情况。结果:获得了稳定表达GFP的转染后细胞;细胞的体外和体内生长未受转染的影响。裸鼠原位种植GFP/A549,利用KODAK IS2000MM确认4只有肝转移,9只有脑转移。HE染色、免疫组化确认4只有肝转移,11只有脑转移,与KODAK IS2000MM检测结果对比差异无统计学意义(P>0.05)。结论:GFP标记人肺腺癌细胞A549原位种植法能更简便的建立NSCLC转移模型;KODAK IS2000MM能够非侵入性和无创性检测肿瘤转移。

Abstract: 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 (P>0.05). 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(P>0.05). Pathological and immunohistochemistry examinations discovered brain metastases were found in 11 mice, liver metastases

Key words: NSCLC, Liver metastasis, Brain metastasis, GFP, Orthotopic transplantation

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