胶质瘤来源外泌体通过高迁移率族蛋白B1促进胶质瘤干细胞形成

摘要 目的：研究胶质瘤来源外泌体中高迁移率族蛋白B1（HMGB1）对胶质瘤干细胞形成的影响及其意义。方法：使用外泌体提取试剂盒提取原代胶质母细胞瘤来源外泌体，通过透射电子显微镜、纳米粒度电位仪和Western blotting对外泌体进行鉴定；采用Western blotting检测外泌体中HMGB1的表达量；通过qRT-PCR、Western blotting、克隆球计数检测外泌体对胶质瘤干细胞形成的影响；siRNA敲低HMGB1的表达水平，并通过qRT-PCR、Western blotting、克隆球计数检测外泌体中HMGB1对胶质瘤干细胞形成的影响。结果：原代胶质瘤细胞可以分泌外泌体到肿瘤微环境并且外泌体中存在HMGB1；原代胶质瘤细胞来源外泌体可以上调邻近胶质瘤细胞干性相关分子CD133、OCT4、NANOG、SOX2的表达并促进干细胞克隆球的形成；通过siRNA敲低原代胶质瘤细胞HMGB1的表达后，外泌体中HMGB1的含量降低并且外泌体促进胶质瘤干细胞形成的作用减弱。结论：胶质瘤细胞来源外泌体可以通过HMGB1促进胶质瘤干细胞的形成。

Glioma-derived Exosomes Promote the Formation of Glioma Stem Cells through High Mobility Group Protein 1

ABSTRACT Objective: To investigate the effect of high mobility group protein B1 (HMGB1) in glioma-derived exosomes on the formation of glioma stem cells. Methods: The exosomes derived from primary glioblastoma were extracted using the exosome extraction kit, and the exosomes were identified by transmission electron microscopy, nano particle size potentiometer and Western blotting; Western blotting was used to detect the expression of HMGB1 in exosomes; the effect of exosomes on the formation of glioma stem cells was detected by qRT-PCR, Western blotting, and sphere counting; the expression level of HMGB1 was knocked down by siRNA and the effect of HMGB1 in exosomes on the formation of glioma stem cells was detected by qRT-PCR, Western blotting and sphere counting. Results: Primary glioma cells could secrete exosomes into tumor microenvironment and HMGB1 is present in the exosomes; primary glioma cells-derived exosomes could up-regulate the expression of stem-related genes CD133, OCT4, NANOG, SOX2 in adjacent glioma cells and promote the formation of tumor spheres. After knocking down the expression of HMGB1 in primary glioma cells by siRNA, the content of HMGB1 in exosomes is reduced and the role of exosomes in promoting the formation of glioma stem cells is attenuated. Conclusion: Exosomes derived from glioma cells could promote the formation of glioma stem cells through HMGB1.