摘要
目的探究牙龈卟啉单胞菌来源的外泌体(P.gingivalis-Exo)对食管癌增殖迁移的影响。方法超速离心法提取P.gingivalis-Exo,通过透射电镜、纳米粒度分析仪对外泌体进行物理表征分析;共聚焦显微镜观察红色荧光染料DIR标记的外泌体被食管癌细胞(KYSE-30/KYSE-150)摄取能力;外泌体与食管癌细胞共培养后,采用MTT、Transwell、划痕和Western blot实验评估外泌体干预前后对食管癌细胞增殖、迁移和侵袭以及TGF-β1信号通路相关蛋白表达水平的影响。结果透射电镜观察P.gingivalis-Exo大小均一,形状如典型的外泌体“圆碟”状,P.gingivalis-Exo粒径约为130 nm;DIR标记的外泌体可被食管癌细胞摄取;与对照组相比,外泌体共培养组细胞增殖能力明显增强(P<0.05),侵袭细胞数量明显增多(P<0.05),划痕愈合速度也显著加快(P<0.05);Western blot分析TGF-β信号通路相关蛋白TGF-β1、Smad2表达明显增加。结论TGF-β1/Smad2通路可能参与了P.gingivalis-Exo促进食管癌细胞的增殖、迁移和侵袭。
Objective To investigate the effects of exosomes derived from P.gingivalis on the proliferation and migration of esophageal cancer cells.Methods Exosomes from P.gingivalis were isolated by ultracentrifugation,and morphology and particle size distribution of exosomes were identified by TEM and nanoparticle size analyzer respectively.Confocal microscopy was performed to observe the uptake of DIR-labeled red fluorescent exosomes by esophageal cancer cells(KYSE-30/KYSE-150).After co-culture with exosomes and esophageal cancer cells,MTT,transwell,wound healing and western blot experiments were used to evaluate the effects of exosomes on proliferation,migration and invasion of esophageal cancer cells before and after intervention,as well as the expression levels of TGF-β1-signaling pathway-related proteins.Results The size of P.gingivalis-exo was uniform,shaped like a typical exosome"disc",and the size of P.gingivalis-exo was about 130 nm.Results of confocal microscopy showed that red fluorescent dye DIR-labeled exosomes were taken up by esophageal cancer cells.Compared with the control group,cell proliferation ability of exosome co-culture group was significantly enhanced(P<0.05),the number of invasive cells was significantly increased(P<0.05),the rate of scratch healing was also markedly accelerated(P<0.05).Western blot results showed that TGF-β1,Smad2 expression increased markedly.Conclusion TGF-β1/Smad2 pathway may be involved in P.gingivalis-Exo to promote the proliferation,migration and invasion of esophageal cancer cells.
作者
王浩洁
史浩
齐义军
朱梦玺
陈攀
兰子君
张灏
梁高峰
高社干
WANG Hao-jie;SHI Hao;QI Yi-jun;ZHU Meng-xi;CHEN Pan;LAN Zi-jun;ZHANG Hao;LIANG Gao-feng;GAO She-gan(Medical College,Henan University of Science and Technology,Luoyang 471023,China;Henan Key Laboratory of Cancer Epigenetics,The First Affiliated Hospital School of Clinical Medicine of Henan University of Science and Technology,Luoyang 471023,China;Institute of Oncology Precision Medicine and Pathology,Jinan University Medical College,Guangzhou 510632,China)
出处
《食管疾病》
2019年第4期17-22,共6页
Journal of Esophageal Diseases
基金
国家自然科学基金(81741147)
中国博士后基金(2019M652533)
