摘要
目的探讨肿瘤细胞与成纤维细胞之间是否存在线粒体转移现象,研究发生线粒体转移后对肿瘤细胞抗药性的影响。方法分别采用荧光蛋白标记肿瘤细胞和成纤维细胞线粒体。采用激光共聚焦显微镜观察细胞和线粒体的形态。采用流式细胞仪分析并分选双阳性细胞群体。采用MTT法检测阿霉素对细胞的增殖抑制作用。结果成功构建HCT-15/GFP细胞和NIH3T3/mito-RFP细胞。从两种细胞共培养体系中,分选出GFP+/RFP+双阳性细胞,比率为3.32%±0.58%。HCT-15/GFP+/RFP+对阿霉素的敏感性较HCT-15/GFP显著下降,化疗抗性提高了3.18倍。结论成纤维细胞中的线粒体能向肿瘤细胞进行转移。发生线粒体转移的肿瘤细胞对阿霉素出现了耐药现象。这为阿霉素耐药提出了新的作用机制。
ObjectiveTo explore whether the existance of mitochondrial transference between cancer cells and fibroblast cells,and to study the effect of mitochondrial transference on drug resistance of tumor cells.MethodsThe cell lines were labeled by fluorescent protein.The morphology of cells and mitochondria had been observed by confocal microscopy.The double positive population of fluorescent protein cells was sorted by flow cytometry.MTT assay had been used to test inhibition effect of doxorubicin on proliferation of cells.ResultsHCT-15/GFP cells and NIH3T3/mito-RFP cells had been successfully constructed.The HCT-15/GFP+/GRP+cells were sorted and their ratio was3.32%±0.58%from the co-cultured mixture cells.The sensitivity of HCT-15GFP+/RFP+to doxorubicin was dramatically decreased and the resistance to chemotherapy had been increased3.18fold.ConclusionThe tranference of mitochondria from fibroblast cells to cancer cells can contribute the resistance to chemotherapy.This provide a new mechanism for resistance to doxorubicin.
作者
郭向华
弓建华
GUO Xiang-hua1;GONG Jian-hua(Beijing Institute of Hepatology, Beijing You An Hospital, Capital Medical University, Beijing 100069, China;Institute of Medicinal Biotechnology Chinese Academy of Medical Sciences, Beijing 100050, China)
出处
《临床和实验医学杂志》
2017年第23期2312-2315,共4页
Journal of Clinical and Experimental Medicine
基金
协和青年基金资助项目(编号:33320140092)
关键词
肿瘤细胞
线粒体转移
阿霉素耐药
Tumor cells
Mitochondrial transference
Doxorubicin resistance
作者简介
弓建华,E-mail:ann_gong@hotmail.com
