放射治療是神經膠質瘤的治療中很重要的一個療法。然而對腫瘤施於高劑量的放射治療後，腫瘤卻經常在放射治療照射區域再度復發，使得神經膠質瘤的治癒率非常的低，因此降低腫瘤在放射治療後的復發率是很急需解決的問題。在本實驗室之前的研究指出，放射治療會增加腫瘤相關巨噬細胞與基質衍生細胞因子(SDF-1)的表現增加，進而影響腫瘤的侵襲性。因此本研究主要目的在於進一步探討阻斷SDF-1/CXCR4作用的藥物AMD3100是否能增強放射療效，並觀察骨髓衍生細胞(Bone-marrow derived cells)在放射治療後不同天數上腦腫瘤微環境的變化。結果顯示跟病危時期相比在放射治療後第4天，高 (32 Gy)、 低 (8 Gy) 劑量放射治療組別的單核球CD11b、M2型巨噬細胞CD68聚集程度及血管周邊細胞的密度都有顯著提升，而在高劑量放射治療組血管密度與M1型巨噬細胞F4/80則有顯著的下降。而在藥物治療完後一天，CD68及CD11b在各組別的表現，都比放射治療第4天有著顯著下降的表現，反而在F4/80聚集程度普遍有顯著的增加，但血管密度及血管周圍細胞重合程度大多也維持為顯著下降。在化療藥物及放射治療合併治療組別顯示AMD3100能有效阻斷骨髓衍生細胞進入腫瘤細胞區域，並能顯著的降低血管密度及血管周邊細胞包圍血管的表現。此外，結果顯示使用藥物AM3100本身即可延長動物的存活，而在合併兩種不同高低劑量的放射治療結果顯示只能有效的能增強高劑量的放射治療效果並確實延長動物的存活時間。
此研究顯示腫瘤再度復發會刺激骨髓衍生細胞進入並活化成M2型巨噬細胞CD68幫助腫瘤再度生長。骨髓衍生細胞會隨著腫瘤在經過放射治療後腫瘤微環境的改變被活化成不同腫瘤相關巨噬細胞幫助其生長，更加確定骨髓衍生細胞在腫瘤經過放射治療後在腫瘤細胞區域所扮演的角色。了解骨髓衍生細胞在放射治療後所扮演的角色有助日後對腫瘤復發的治療能有更大的突破，以期達到更加腫瘤的治療效果。

Radiotherapy plays a major role in the treatment of glioblastoma multiforme (GBM).The high doses of radiation delivered in the treatment of patients with GBM, the tumors invariably recur within the irradiation field, resulting in a low cure rate. Therefore, reducing the recurrence rate after radiation therapy is urgent problem. Lab early study has demonstrated that radiation-induced brain tumor invasiveness is associated with the level of SDF-1 production by tumor cells. This study further examined if AMD3100, a drug to block the interaction of SDF-1 with its receptor CXCR4, could enhance the efficacy of radiation therapy and explored the change of tumor microenvironments after radiotherapy in a murine astrocytoma tumor model, ALTS1C1. Our first result demonstrated that the administration of AMD3100 could prolong the surviving time of tumor-bearing mice after 32 Gy of radiation therapy, but not 8 Gy of radiation therapy. The immunohistochemical (IHC) staining study further showed that the number of CD11b+ TAMs or CD68+ M2 macrophages, and the density of pericyte cells were significantly increased at 4 days after radiation therapy, but the microvascular density (MVD) and the number of F4/80+ M1 macrophages were significantly decreased. The administration of AMD3100 s significantly decreased the number of CD68+ or CD11b+ macrophages, but increased the number of F4/80+ macrophages. The MVD and the pericyte coverage rate were also decreased significantly. In summary, this study shows that tumor recurrence stimulates the bone marrow-derived cells to enter and the CD68 positive activated alternative M2 macrophages help regrowth of the tumor and the administration of AMD3100following radiotherapy can effectively block the recruitment of bone marrow-derived cells, reduce the density of blood vessels and pericyte cell coverage, and enhance the efficacy of high dose of radiation therapy for brain tumor.

內容
摘要 2
Abstract 3
致謝 4
1.緒論 6
1.1 癌症 6
1.2 神經膠質瘤(Gliomas) 6
1.3 腫瘤微環境(Tumor microenvironment) 7
1.3.2 免疫調控 8
1.4骨髓衍生細胞與單核球 8
1.5 AMD3100(普樂沙福, Plerixafor) 10
1.5.1 AMD3100之副作用 10
2.材料與方法 12
2.1細胞培養 12
2.2 動物實驗 12
3.實驗結果 18
3.探討治療後小鼠存活率的增加 18
3.2.探討放射治療後小鼠腦腫瘤內單核球CD11b的變化 18
3.3探討M 2型巨噬細胞在放射治療之腦腫瘤微環境所扮演的角色 22
3.4探討M 1型巨噬細胞F4/80在放射治療之腦腫瘤微環境所扮演的角色 24
3.6血管中平滑肌細胞的觀察 31
4.討論 32
5.圖表 36
6.引用文獻 76

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