肺癌A549裸鼠移植瘤模型高效建立方法的应用

Exploration and application of effective method to establish nude mice xenograft model of human lung cancer cells A549

目的:探讨裸鼠高效移植瘤模型的建立方法,为进行肿瘤的体内相关研究和抗癌药物的筛选提供实验基础。方法:肺癌A549细胞接种于裸鼠皮下,出瘤后制备单细胞悬液和组织块分别接种和塞入皮下,观察初次培养细胞成瘤组、二次单细胞悬液成瘤组和二次组织块成瘤组的出瘤率和移植瘤的生长情况,PI单染流式细胞仪检测细胞周期的变化,细胞毒实验采用MTT法。结果:3组中二次组织块成瘤组的出瘤率显著高于另外两组,细胞增殖块,移植瘤的变异小;细胞周期分析显示二次组织块成瘤组的细胞S和G2/M期比例增高,细胞增殖指数显著高于另外两组;但细胞毒MTT的实验结果显示3组肿瘤细胞对化疗药物顺铂的敏感性差异无显著性(P>0.05)。结论:二次组织块成瘤可显著提高A549的成瘤率,且不影响肿瘤细胞对化疗药物的反应性,是一种高效的裸鼠移植瘤模型建立方法。

Objective：To study an effective method to establish xenograft model in nude mice.Methods：Human cancer A549 cells were subcutaneously inoculated in nude mice.After tumor formation,the single cells suspension or mini-tissue samples were prepared,inoculated and subcutaneously imbedded.Three kinds of different tumor samples were subcutaneously transplanted into nude mice.The tumor formation rate and xenograft growing characteristics were observed and analyzed.PI staining and subsequent flow cytometric analysis was used to analyze cell cycle.The cells viability was determined by MTT assay.Results：Mini-tissue samples significantly improved the xenograft formation.The percent of S and G2/M phase cells was higher in mini-tissue samples derived heterogeneous tumors by flow cytometric analysis.However,MTT results demonstrated that the cytotoxic activity of cisplatin against foregoing three group cells did not show significant difference.Conclusion：The tumor formation rate is significantly improved by mini-tissue samples without affecting the sensibility of chemotherapeutic cisplatin.Transplanting with mini-tissue samples is an effective method to establish the model of nude mice xenograft.