The Study Of Macrophages Fuse With Glioma Cells And Promote Glioma Cell Metastasis And Invasion

Central nervous system tumors cause serious damage to people’s health and life, among which the most common seen is the glioma. Malignant glioma has strong invasion abitily, and is difficult to be eradicated completely through neurosurgery operation. The subsequent treatment with radiotherapy and chemotherapy is hard to completely eliminate the residual tumor cells, which eventually lead to the recurrence of the glioma, causing patients’ poor prognosis and high death rate. Therefore, patients with malignant glioma survive less than 2 years in average.In recent years, with the introduction of the cancer stem cell theory, correlational researchers begin to survey the malignant gliomas and its biological characteristics again. Several studies have shown that the malignant glioma contains glioma stem cells, which are of low content but have super strong self-renewal and multi-differentiation capacity, and play a very important role in the occurrence and development of the tumor. Therefore, the research of the malignant gliomas’ biological characteristics has profound significance on the acquaintance of the malignant gliomas’ invasive growth. In addition, numerous study results have shown that the inflammatory cells are widely involved in the occurrence and development of the tumor. The immune microenvironment of the malignant gliomas and its infiltrating immune cells both take part in the processes of the tumor angiogenesis, the immune escape and the invasion and metastasis. Among the tumor infiltrating immune cells, the tumor-associated macrophages is the most important subgroup and makes up the highest proportion. However, relatively few numbers of studies on the relationship between macrophages and glioma cells are made at this stage, so its role in the invasion progress of glioma is still unclear. Besides, whether the macrophage cells can be fused with glioma in order to produce the biological effect lacks research.In the first part of this study, the author attempts to infect the GL261 mice glioma cells and the RAW264.7 mice macrophage with lentivirus Lv-RFP and pAOV.SYN.GFP, flow cytometry sorting each positive cells, successfully establishing the GL261 mice glioma cell with stable expression of red fluorescent protein(RFP) and the RAW264.7 mouse macrophage with stable expression of green fluorescent protein(GFP); Live cell workstation monitors that the 24 h co-culture of GL261-RFP and RAW264.7-GFP shows the phenomenon of spontaneous fusion and the presence of a split and the fusion efficiency improves 5 times with the PEG-induce. This part establishes the foundation of further discussion of the tumor stem cell formation with the fusion of macrophages and glioma cell and its role in the invasion. In the second part, the author adopts the flow sorting technique to isolate the CD133+ glioma cell and CD133- glioma cell from the mice transplanted tumor model, and co-cultures them with macrophages, and later carries out the experiment of glioma cell migration and invasion. The result shows that the migration ability of CD133+ glioma cell and CD133- glioma, which are the source of GL261 transplantation tumor, has no obvious difference, and the co-culture with macrophages does no significant change to the invasion ability of both the cells. Compared with CD133- glioma cell, CD133+ glioma cell has stronger invasion ability. After the co-culture with macrophages, the invasion ability of both CD133- glioma cell and CD133+ glioma cell improves largely. And compared with CD133- glioma cell, the invasion ability of the CD133+ glioma cell improves more obviously.