本文關(guān)鍵詞： 膠質(zhì)瘤 成纖維細胞生長因子受體 NVP-BGJ398 血管生成擬態(tài)　出處：《南方醫(yī)科大學》2017年碩士論文　論文類型：學位論文

【摘要】：研究背景:膠質(zhì)細胞瘤是常見的中樞神經(jīng)系統(tǒng)惡性腫瘤,患者的半數(shù)生存期不足兩年。成纖維細胞生長因子受體(FGFR)是一類酪氨酸激酶膜受體。FGFR信號通路涉及腫瘤細胞的各種活動,如腫瘤的增殖,血管生成,侵襲及腫瘤干細胞多功能性的維持等。文獻顯示FGFR在膠質(zhì)瘤中表達上升,提示FGFR是治療膠質(zhì)瘤的潛在靶點。NVP-BGJ398是目前可獲得的強效選擇性抑制FGFR的藥劑。血管生成擬態(tài)是一種與血管相似的官腔結(jié)構(gòu),但它的管壁不是由血管內(nèi)皮細胞構(gòu)成的而是由腫瘤細胞組成的,這個官腔可以給膠質(zhì)瘤提供額外的血供,促進惡性腫瘤的發(fā)展。目的:檢測FGFR抑制劑NVP-BGJ398對抑制膠質(zhì)瘤細胞增殖、遷移和侵襲的影響;檢驗NVP-BGJ398對膠質(zhì)瘤細胞形成血管生成擬態(tài)的影響;探索NVP-BGJ398影響膠質(zhì)瘤細胞的機制。方法:1、進行WB實驗和免疫組化染色分析蛋白表達;2、運用增殖實驗、平板克隆實驗和皮下移植瘤模型檢測膠質(zhì)瘤細胞生長;3、進行侵襲實驗和遷移實驗分析腫瘤細胞的侵襲和遷移能力;4、運用小管生成實驗和CD34/PAS雙染檢測腫瘤細胞血管擬態(tài)的形成;5、統(tǒng)計分析。結(jié)果:1、Western blotting顯示NVP-BGJ398處理可以有效的抑制膠質(zhì)瘤細胞中的FGFR磷酸化激活成為pFGFR和降低膠質(zhì)瘤細胞中Vimentin、MMP2和MMP14 的表達(P0.05);2、CCK-8實驗結(jié)果表明經(jīng)過NVP-BGJ398的處理兩株細胞的增殖速度明顯的下降(P0.05);平板克隆實驗對照組中克隆形成數(shù)量遠大于經(jīng)過NVP-BGJ398 實驗組(P0.05);3、劃痕實驗中對照組U251MG細胞遷移速度明顯快于NVP-BGJ398實驗組(P0.05);Transwell遷移和侵襲實驗結(jié)果表明穿膜細胞數(shù)在NVP-BGJ398處理后都明顯的減少(P0.05);4、小管形成實驗中NVP-BGJ398實驗組中膠質(zhì)瘤細胞小管周長和交點數(shù)較對照組明顯的減少;5、U87MG皮下成瘤實驗中NVP-BGJ398實驗組(0.19±0.06g)腫瘤重量明顯輕于對照組(0.45±0.11g)(P0.05);6、CD34/PAS雙染VM計數(shù)結(jié)果顯示皮下瘤模型NVP-BGJ398實驗組中VM的數(shù)量(13.85±3.96)較對照組(26.40±5.06)明顯的減少(P0.05);7、免疫組織染色顯示NVP-BGJ398實驗組中Vimentin、MMP2和MMP14平均光密度(2.50±0.36,1.89±0.95,1.26±0.15)明顯淺于對照組(4.13±1.15,4.90±0.52,3.77±0.39)(P0.05)。結(jié)論:在體內(nèi)外實驗中發(fā)現(xiàn)FGFR抑制劑NVP-BGJ398具有良好的抗膠質(zhì)瘤作用。NVP-BGJ398除了抑制腫瘤細胞的增殖、遷移以及侵襲,還具有抑制血管生成擬態(tài)生成的作用。其機制與能夠抑制FGFR的激活下調(diào)vimentin、MMP14和MMP2的表達有關(guān)。提示成纖維細胞生長因子受體是治療膠質(zhì)瘤的一個有效靶點。
[Abstract]:Background: glioma is a common malignant tumor of the central nervous system. Fibroblast growth factor receptor (FGFR) is a type of tyrosine kinase receptor. FGFR signaling pathway involves various activities of tumor cells, such as tumor proliferation, angiogenesis, Invasion and multifunctional maintenance of tumor stem cells. The literature shows that the expression of FGFR is increased in gliomas. It is suggested that FGFR is a potential target for the treatment of glioma. NVP-BGJ398 is a potent and selective inhibitor of FGFR. However, its wall is composed not of vascular endothelial cells but of tumor cells, which can provide additional blood supply to glioma and promote the development of malignant tumor. Objective: to detect the inhibitory effect of FGFR inhibitor NVP-BGJ398 on the proliferation of glioma cells. The effects of NVP-BGJ398 on the angiogenesis of glioma cells were examined. The mechanism of NVP-BGJ398 affecting glioma cells was explored. Plate clone assay and subcutaneous tumor transplantation model were used to detect the growth of glioma cells. Invasion and migration experiments were carried out to analyze the invasion and migration ability of tumor cells. Microtubule formation test and CD34/PAS double staining were used to detect tumor blood vessels. Results NVP-BGJ398 treatment could effectively inhibit the activation of FGFR phosphorylation into pFGFR in glioma cells and decrease the expression of Vimentinine MMP2 and MMP14 in glioma cells. The results showed that NVP-BGJ398 treatment could effectively inhibit the activation of FGFR phosphorylation into pFGFR and decrease the expression of Vimentinine MMP2 and MMP14 in glioma cells. The proliferation rate of the two cells decreased significantly (P 0.05), the number of clone formation in the control group was much larger than that in the NVP-BGJ398 experimental group, and the migration rate of U251MG cells in the scratch test group was significantly faster than that in the NVP-BGJ398 experimental group. The results showed that the number of perforating cells decreased significantly after NVP-BGJ398 treatment. In the tubule formation test, the circumference and intersection number of glioma cells in the NVP-BGJ398 experimental group were significantly reduced compared with those in the control group. In the subcutaneous tumorigenesis experiment of U87MG, the number of the glioma cells in the NVP-BGJ398 experimental group was significantly reduced. Tumor weight (0.19 鹵0.06g) was significantly lighter than that in control group (0.45 鹵0.11g). The results of VM counting of CD34 / pas double staining showed that the number of VM in the subcutaneous tumor model NVP-BGJ398 group was 13.85 鹵3.96, significantly lower than that in the control group (26.40 鹵5.06g). The immunohistochemical staining showed that Vimentinine MMP2 and MMP14 average optical density in NVP-BGJ398 experimental group were 2.50 鹵0.36 鹵1.89 鹵51.26 鹵0.15). Conclusion: in vivo and in vitro, FGFR inhibitor NVP-BGJ398 has a good anti-glioma effect. NVP-BGJ398 can not only inhibit the proliferation of tumor cells, but also inhibit the proliferation of tumor cells. Migration and invasion also inhibit angiogenesis mimicopoiesis. The mechanism is related to the down-regulation of FGFR activation and down-regulation of the expression of MMP14 and MMP2. It suggests that fibroblast growth factor receptor is an effective target for the treatment of glioma.
【學位授予單位】：南方醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R739.41

【參考文獻】

相關(guān)期刊論文 前1條

1 武世伍;俞嵐;承澤農(nóng);宋文慶;周蕾;陶儀聲;;Expression of Maspin in Non-small Cell Lung Cancer and Its Relationship to Vasculogenic Mimicry[J];Journal of Huazhong University of Science and Technology(Medical Sciences);2012年03期

，

本文編號：1503254