exosomes在肺癌细胞对顺铂敏感性降低过程中的作用研究

Effect of exosomes derived from lung cancer cells under cisplatin condition on sensitivity of cells to cisplatin

目的研究肺癌细胞分泌的exosomes在同源细胞对顺铂的敏感性调节过程中的可能作用。方法采用超速离心和Exoquick-TC结合的方法从肺癌细胞系A549和H1975的上清液中分离出exosomes,透射电子显微镜观察其形态,western blot检测其CD63蛋白表达;共聚焦显微镜观察细胞胞吞exosomes过程;分别提取顺铂处理肺癌细胞来源exosomes和未处理肺癌细胞来源exosomes,并使用2种exosomes分别处理肺癌细胞,采用CCK-8法检测上述获得的exosomes对肺癌细胞顺铂敏感性的影响。结果透射电子显微镜下观察肺腺癌细胞来源的exosomes具有特征性的盘状结构,直径30-100 nm,western blot结果显示exosomes富含CD63蛋白;显微镜下可观察到exosomes可进入细胞;同时经顺铂处理过肺癌细胞分泌的exosomes可降低同源细胞对顺铂的敏感性。结论减少exosomes的分泌和传递可能会增加顺铂化疗的疗效,这为肺癌的治疗提供了一种新的思路。

Objective To study whether the sensitivity of lung cancer cells to cisplatin could be influenced by exosomes derived from the cells. Methods Exosomes were isolated from the supernatant of lung cancer cells with the method of ultracentrifugation and Exoquick-TC in combination. The morphology were observed with transmission electron microscopy, and the protein expression of CD63 were detected with western blot. The state of the cells treated with exosomes was observed with confocal microscopy. Exosomes were extracted from supernatant of lung cancer ceils cultured in cisplatin condition or normal condition respectively, which were applied to the cells and the cell viability was detected with CCK-8. Results Lung adenocarcinoma cell-derived exosomes showed discoid structure with a double membrane , and a diameter of 30-100 nm. Exosomes were enriched CD63 protein and could be observed to enter ceils under the microscope. The exosomes obtained under cisplatin condition could reduce the sensitivity of cells to cisplatin. Conclusions Reducing the secretion and transmission of exosomes might increase the efficacy of cisplatin, which provides a new idea about the treatment of lung cancer.