【作者】 樊婷婷； 赵毅； 李娟； 张敬； 王福金； 王爱国； 王靖宇；

【Author】 FAN Ting-ting;Zhao Yi;LI Juan;Zhang Jing;Wang Fu-jin;Wang Ai-guo;Wang Jing-yu;Laboratory animal center,Dalian Medical University;

【机构】 大连医科大学实验动物中心；

【摘要】 目的建立Hras12V癌基因诱导的小鼠肝肿瘤细胞系,为携带Ras癌基因肝肿瘤细胞的生物学特性及治疗研究提供生物学材料。方法选取9月龄Hras12V转基因雄鼠一只,采取肝肿瘤组织,通过组织培养的方法获取肝肿瘤细胞系。并通过PCR、Cell cycle以及Western Blot等一系列实验鉴定该转基因小鼠肝癌细胞系的基因型和细胞生物特性。结果通过肝肿瘤组织原代培养的方法获取原代肝肿瘤细胞后,经反复传代最终获得了来自Hras12V转基因雄鼠的稳定肝肿瘤细胞系MMHRL3。该细胞系的细胞形态较为一致,增殖状态良好。分子生物学检测结果表明,该细胞系携带Hras12V转基因,具有肿瘤细胞特性。而且MMHRI3细胞中MEK/ERK和AKT/mTOR信号通路激活。这些生物学特性与Hras12V转基因小鼠的肝肿瘤特性相同。结论本实验室成功建立了Hras12V转基因肝癌雄鼠的肝肿瘤细胞系。MMHRL3细胞系为肝肿瘤发生发展的机制研究及靶向MEK/ERK和.AKT/mTOR信号通路的分子药物的开发提供了良好的研究对象。更多还原

【Abstract】 ObjectiveTo establish a mouse hepatic tumor cell line induced by Hras12 V oncogene for providing biological materials to study the biological characteristics of hepatic tumor cells harboring ras oncogene and related therapy strategies.MethodsA 9 month-old Hras12 V transgenic male mice was sampled and the liver tumor cell lines were obtained by primary tissue culture method.The genetic and basic biological characteristics of the transgenic mice hepatic tumor cell line were identified by PCR,Cell cycle,and Western Blot.ResultsAfter the primary hepatic tumor cells were obtained by the primary tissue culture method,the stable hepatic tumor cell line of Hras12 V transgenic male mice was obtained by repeated passages.The morphology of MMHRL3 cell line was uniform and the proliferation was in good condition.Molecular biological tests showed that the cell line carried the Hras12 V transgene,and have the tumor cell characteristic.In addition,MMHRL3 cell line cells effectively activated MEK/ERK and AKT/mTOR signaling pathway.These biological characteristics were the same as those of the liver tumors of Hras12 V transgenic mice.Conclusion We successfully established a hepatic tumor cell line induced by Hras12 V oncogene.MMHRL3 cell line provides a good research object for the mechanism researches related to liver tumors and the development of molecular drugs targeting MEK/ERK and AKT/mTOR signaling pathway.更多还原