乳腺癌细胞来源的外泌体对乳腺癌BT549细胞增殖、侵袭和迁移的影响  

EFFECTS OF BREAST CANCER-DERIVED EXOSOMES ON PROLIFERATION,INVASION,AND MIGRATION OF BREAST CANCER BT549 CELLS

作　　者：王军[1] 郑文祥 周娜[1] 高帅 侯琳[1] WANG Jun;ZHENG Wenxiang;ZHOU Na;GAO Shuai;HOU Lin(Department of Biochemistry and Molecular Biology, School of Basic Medicine, Qingdao University, Qingdao 266021, China)

机构地区：[1]青岛大学基础医学院生物化学与分子生物学系,山东青岛266021 [2]青岛大学附属妇女儿童医院心脏中心

出　　处：《精准医学杂志》2021年第1期33-37,共5页Journal of Precision Medicine

基　　金：国家自然科学基金资助项目(81472542);山东省重点研发项目(2019GSF107025)。

摘　　要：目的研究乳腺癌细胞来源的外泌体对乳腺癌BT549细胞的增殖、侵袭和迁移的影响。方法使用超速离心的方法对人乳腺癌BT549细胞培养液中的外泌体进行提取和纯化,以通用透射电子显微镜观察外泌体的形态和大小,蛋白免疫印迹实验鉴定外泌体的标志蛋白,以CCK-8实验、细胞划痕实验和Transwell实验检测外泌体对BT549细胞增殖、侵袭和迁移能力的影响。结果透射电子显微镜下外泌体的形状为椭圆形,外包被一层磷脂膜,直径40~110 nm;蛋白免疫印迹实验结果显示外泌体内标志蛋白TSG101以及外标志蛋白CD63均明显富集;CCK-8实验结果显示共培养24 h后,实验组的BT549细胞增殖力较对照组明显增强(F=12.99~50.95,P<0.001);细胞划痕实验结果显示外泌体明显促进了BT549细胞的迁移(t=8.93,P<0.01);Transwell实验结果显示实验组BT549细胞迁移数较对照组明显增多(t=4.77,P<0.01)。结论乳腺癌细胞来源的外泌体能促进肿瘤细胞的增殖、侵袭和迁移,表明肿瘤来源外泌体可作为交叉信号的传递平台,在促进肿瘤的生长和肿瘤转移方面发挥自分泌作用。Objective To investigate the effects of breast cancer-derived exosomes on the proliferation,invasion,and migration of breast cancer BT549 cells.Methods The exosomes from human breast cancer BT549 cell culture medium were extracted and purified by ultracentrifugation.The morphology and size of the exosomes were assessed by universal transmission electron microscopy.Marker proteins of the exosomes were identified by Western blotting.Cell Counting Kit-8(CCK-8)assay,wound healing assay,and Transwell assay were used to determine the effects of exosomes on the proliferation,invasion,and migration of BT549 cells.Results Under a transmission electron microscope,the exosomes were elliptic,with a diameter of 40-110 nm,covered by a phospholipid membrane.Western blotting showed that both the interior marker protein TSG101 and the exterior marker protein CD63 in the exosomes were significantly enriched.The CCK-8 assay showed that the proliferation of BT549 cells in the experimental group was significantly increased as compared with that in the control group after 24 h of co-culture(F=12.99-50.95,P<0.001);the wound healing assay showed that exosomes significantly promoted the migration of BT549 cells(t=8.93,P<0.01);the Transwell assay showed that the number of migrating BT549 cells in the experimental group was significantly increased than that in the control group(t=4.77,P<0.01).Conclusion Breast cancer-derived exosomes can promote the prolife-ration,invasion,and migration of tumor cells,suggesting that tumor-derived exosomes can play an autocrine role in promoting tumor growth and metastasis by acting as a cross-signal transmission platform.

关 键 词：乳腺肿瘤 细胞系 肿瘤 外泌体 细胞增殖 细胞运动 肿瘤侵润 体外研究 

分 类 号：R737.9[医药卫生—肿瘤]