杨毅 张志军

【摘 要】目的：探究肺癌细胞分泌的外泌体对顺铂的敏感性调节。方法：通过超速离心从肺癌细胞A549上清液中分离得到外泌体，采用透射电子显微镜（TEM）来观察外泌体的形貌，蛋白质印迹法检测CD63和CD81的蛋白表达；将肺癌细胞来源外泌体一组用顺铂处理，另一组不进行处理，将2组外泌体与肺癌细胞共同培养，然后用CCK-8法来检测外泌体在同源细胞中对顺铂的敏感性调节过程中的作用。结果：外泌体在透射电子显微镜下观察到具有盘状的形貌，直径约30-100nm；蛋白质印迹法显示外泌体富含CD63和CD81蛋白；未顺铂处理的exosome干预细胞组较PBS处理的细胞组细胞存活率高4%，而顺铂处理的exosome干预细胞组较未顺铂处理的exosome干预细胞组细胞存活率高24%，P<0.05。结论：肺癌细胞来源的顺铂处理的外泌体降低了肺癌细胞对顺铂的敏感性，这可能为肺癌的化疗提供一种新的靶点和思路。

【關键词】肺癌；外泌体； 化疗

【中图分类号】R197 【文献标识码】A 【文章编号】1004-7484（2019）02-0091-02

Lung cancer cell-derived cisplatin-treated exosomes reduce the sensitivity of lung cancer cells to cisplatin

Yang yi Zhang zhi jun（Corresponding author）

（Center for reproductive medicine， Taihe hospital， Shiyan442000， Hubei， China）

【Abstract】Objective：To investigate the sensitivity of exosomes secreted by lung cancer cells to cisplatin. Methods：Exosomes were isolated from the supernatant of lung cancer cell A549 by ultracentrifugation. The morphology of exosomes was observed by transmission electron microscopy （TEM）. The protein expression of CD63 and CD81 was detected by Western blotting. One group was treated with cisplatin and the other group was not treated. Two groups of exosomes were co-cultured with lung cancer cells， and then CCK-8 method was used to detect the sensitivity of exosomes to cisplatin in homologous cells. The role of the adjustment process. Results：Exosomes were observed to have a disk-like morphology under transmission electron microscopy with a diameter of about 30-100 nm. Western blotting showed that exosomes were rich in CD63 and CD81 proteins. The non-cisplatin-treated exosome intervention cell group had a 4% higher cell viability than the PBS-treated cell group， while the cisplatin-treated exosome intervention cell group had a 24% higher cell viability than the non-cisplatin-treated exosome intervention cell group， P < 0.05. Conclusion：Lung cancer cell-derived cisplatin-treated exosomes reduce the sensitivity of lung cancer cells to cisplatin， which may provide a new target and idea for chemotherapy of lung cancer.

【Key word】Lung cancer； Exosome； Chemotherapy

肺癌的发病率和致死率在所有恶性肿瘤中居于首位，并且逐年上升，严重危害着人民健康。肺癌起病隐匿，病情发展迅速，影响肺癌预后的最主要因素是分期，约75%～85%的肺癌患者在诊断时就已属晚期，失去彻底切除机会[1-3]。外泌体（exosome，Exo）是由多种细胞如树突状细胞、肿瘤细胞等经内吞逆出芽形成许多小囊泡，直径约30～100nm。它可以在不直接接触的情况下完成细胞间生物信号传导[4]。本研究成功从肺癌细胞中提出外泌体，并揭示了肺癌细胞来源的外泌体能调节肺癌细胞对顺铂化疗敏感性，这为肺癌的治疗提供了一种新的思路和靶点

1 资料与方法

1.1 细胞培养 肺癌细胞A549 购于上海细胞库。……