肺癌干细胞分泌外泌体增强肺癌细胞对化疗敏感性的研究

Study on the effects of Exosomes derived from lung cancer stem cells to enhance the chemo-sensitivity of lung cancer cells

目的研究化疗药物诱导后肺癌干细胞分泌外泌体(Exosomes)对肺癌细胞化疗敏感性的影响。方法采用流式荧光细胞分选技术从人肺癌细胞系A549中分离出肺癌干细胞,采用1μmol/L吉非替尼培养液诱导肺癌干细胞分泌外泌体并提取。将外泌体作用于人肺癌细胞系A549,噻唑蓝(MTT)法检测干预前后肺癌细胞的增殖能力和对化疗药物的敏感程度。Western blot检测细胞耐药相关蛋白表达水平。结果采用超速离心方法能提取化疗药物诱导后肺癌干细胞分泌的外泌体;MTT法显示这类外泌体能够促进人肺癌细胞系A549的增殖,导致人肺癌细胞系A549对吉非替尼的IC_(50)值明显增大(P<0.05)。化疗药物诱导后肺癌干细胞分泌外泌体能增强肺癌细胞MDR-1、ABCG2和LRR耐药相关蛋白的表达。结论化疗药物诱导后肺癌干细胞来源的外泌体能增强肺癌细胞增殖活性,并诱导肺癌细胞对化疗药物的耐药。

Objective It is to investigate the effects of Exosomes derived from lung cancer stem cells which induced by chemotherapeutic drugs on the chemo-sensitivity of lung cancer cells. Methods Lung cancer stem cells were isolated by fluorescence activated cell sorting (FACS) from human lung adenocarcinoma cell line A549. The 1umol/L gefitinib culture medium was used to induce lung cancer stem cells to secrete Exosomes. Then the Exosomes were extracted and used to effect lung cancer cell line A549. MTT method was used to evaluate the capability of cell proliferation and chemoresistance. Western Blot was used to detect the expression level of drug resistance related proteins. Results Ultracentrifugation could extract Exosomes from lung cancer stem cells induced by chemotherapeutic drugs. MTT showed that Exosomes could promote the proliferation of human lung cancer cell line A549 and make the IC 50 value of gefitinib increased significantly( P <0.05). Western Blot detection showed that the secretion of Exosomes from lung cancer stem cells induced by chemotherapeutic drugs could enhance the expression of MDR-1, ABCG2 and LRR in lung cancer cells. Conclusion The Exosomes derived from lung cancer stem cells which induced by chemotherapeutic drugs could promotes lung cancer cell proliferation and induces chemotherapeutic drug resistance of lung cancer cells.