摘要
目的:研究缝隙连接蛋白43(CX43)对胶质母细胞瘤(S24-GBM)细胞间网络连接的影响及其在GBM放射抵抗中的作用。方法:采用基因敲除S24-GBM干细胞(S24-GBMSCs)表面CX43的表达或生胃酮(CBX)特异性阻断CX43的功能后,用多光子激光扫描显微镜观察S24-GBMSCs移植裸鼠大脑中肿瘤细胞表面微管(TMs)的形成和小分子物质(Ca^2++和SR101)在细胞间的传递。采用MRI检测放射对GBM体积变化的影响,记录和分析裸鼠生存时间。结果:与对照组相比,敲除CX43基因后S24-GBMSCs的TMs明显缩短,TMs参与形成的网络连接减少;细胞间Ca^2+同步性和SR101扩散能力下降,肿瘤体积缩小,荷瘤裸鼠生存时间显著延长(P均<0.01);CBX可以抑制Ca^2+和SR101的扩散,延缓GBM的生长,但不影响TMs的形成(P>0.05)。采用CBX阻断TMs形成缺陷的S24-GBMSCs表面CX43的功能可以进一步增强上述效应,提高S24-GBM对放射的敏感性(P均<0.05)。结论:CX43参与S24-GBM细胞间网络连接的形成,影响细胞间信号分子的传递,在S24-GBM放射抵抗中起着重要作用。
Objective To investigate the impact of connexin 43(CX43)on the connection of S24 glioblastoma multiforme(S24-GBM)cellular network and to explore its role on radio-resistance.Methods Specific lentiviral vectors were used to knockout CX43 in S24-GBM stem cells(S24-GBMSCs).Alternatively,carbenoxolone(CBX)was used to block transmission of CX43.Subsequently,the animal subjects grafted with S24-GBMSCs were monitored under a multiphoton laser scanning microscope(MPLSM).Dynamic changes of tumor microtubes(TMs)and transmission of Ca^2+and SR101 in the cellular network were recorded.To study the radiosenstivity of S24-GBM before and after CX43 inhibition,MRI scanning of the brains was taken before and after radiation to assess the tumor sizes.Survival time of each subject was also recorded.Results In comparison with control group,knockout of CX43 in S24-GBMSCs led to shorter TMs,less TM connected cells,lower Ca^2+synchronicity and SR101 fluorescence,as well as decreased tumor sizes and prolonged survival time(all P<0.01),which were independent from radiation.However,CBX only demonstrated inhibition on the growth of tumors and the diffussion of Ca^2+and SR101,without affecting TMs formation.These above-mentioned alterations could be enhanced by the combination of gap43 knockout in S24-GBMSCs with blockage of CX43 by CBX(all P<0.05).Conclusion CX43 plays a critical role in the radioresistance of S24-GBM by influencing the formation of S24-GBM cellular network and the transmission of important signaling molecules including Ca^2+and SR101.
作者
黄露露
刘胜文
张孟贤
于世英
邱红
胡国清
Huang Lulu;Liu Shengwen;Zhang Mengxian;Yu Shiying;Qiu Hong;Hu Guoqing(Department of Oncology,Tongji Hospital,Tongji Medical College of Huazhong University of Science and Technology,Wuhan 430030,China;Department of Neurosurgery,Tongji Hospital,Tongji Medical College of Huazhong University of Science and Technology,Wuhan 430030,China)
出处
《中华放射肿瘤学杂志》
CSCD
北大核心
2020年第5期383-387,共5页
Chinese Journal of Radiation Oncology
基金
国家自然科学基金项目(81772680)
华中科技大学双一流自主创新学科医师资助项目(3011540024,5001540074,5001540095)
武汉市中青年医学骨干人才培养工程(2016whzqnyxggrcl)。
