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

【摘要】：研究背景:膠質(zhì)細(xì)胞瘤是常見(jiàn)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(xì)胞生長(zhǎng)因子受體(FGFR)是一類酪氨酸激酶膜受體。FGFR信號(hào)通路涉及腫瘤細(xì)胞的各種活動(dòng),如腫瘤的增殖,血管生成,侵襲及腫瘤干細(xì)胞多功能性的維持等。文獻(xiàn)顯示FGFR在膠質(zhì)瘤中表達(dá)上升,提示FGFR是治療膠質(zhì)瘤的潛在靶點(diǎn)。NVP-BGJ398是目前可獲得的強(qiáng)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(xì)胞構(gòu)成的而是由腫瘤細(xì)胞組成的,這個(gè)官腔可以給膠質(zhì)瘤提供額外的血供,促進(jìn)惡性腫瘤的發(fā)展。目的:檢測(cè)FGFR抑制劑NVP-BGJ398對(duì)抑制膠質(zhì)瘤細(xì)胞增殖、遷移和侵襲的影響;檢驗(yàn)NVP-BGJ398對(duì)膠質(zhì)瘤細(xì)胞形成血管生成擬態(tài)的影響;探索NVP-BGJ398影響膠質(zhì)瘤細(xì)胞的機(jī)制。方法:1、進(jìn)行WB實(shí)驗(yàn)和免疫組化染色分析蛋白表達(dá);2、運(yùn)用增殖實(shí)驗(yàn)、平板克隆實(shí)驗(yàn)和皮下移植瘤模型檢測(cè)膠質(zhì)瘤細(xì)胞生長(zhǎng);3、進(jìn)行侵襲實(shí)驗(yàn)和遷移實(shí)驗(yàn)分析腫瘤細(xì)胞的侵襲和遷移能力;4、運(yùn)用小管生成實(shí)驗(yàn)和CD34/PAS雙染檢測(cè)腫瘤細(xì)胞血管擬態(tài)的形成;5、統(tǒng)計(jì)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(xì)胞中的FGFR磷酸化激活成為pFGFR和降低膠質(zhì)瘤細(xì)胞中Vimentin、MMP2和MMP14 的表達(dá)(P0.05);2、CCK-8實(shí)驗(yàn)結(jié)果表明經(jīng)過(guò)NVP-BGJ398的處理兩株細(xì)胞的增殖速度明顯的下降(P0.05);平板克隆實(shí)驗(yàn)對(duì)照組中克隆形成數(shù)量遠(yuǎn)大于經(jīng)過(guò)NVP-BGJ398 實(shí)驗(yàn)組(P0.05);3、劃痕實(shí)驗(yàn)中對(duì)照組U251MG細(xì)胞遷移速度明顯快于NVP-BGJ398實(shí)驗(yàn)組(P0.05);Transwell遷移和侵襲實(shí)驗(yàn)結(jié)果表明穿膜細(xì)胞數(shù)在NVP-BGJ398處理后都明顯的減少(P0.05);4、小管形成實(shí)驗(yàn)中NVP-BGJ398實(shí)驗(yàn)組中膠質(zhì)瘤細(xì)胞小管周長(zhǎng)和交點(diǎn)數(shù)較對(duì)照組明顯的減少;5、U87MG皮下成瘤實(shí)驗(yàn)中NVP-BGJ398實(shí)驗(yàn)組(0.19±0.06g)腫瘤重量明顯輕于對(duì)照組(0.45±0.11g)(P0.05);6、CD34/PAS雙染VM計(jì)數(shù)結(jié)果顯示皮下瘤模型NVP-BGJ398實(shí)驗(yàn)組中VM的數(shù)量(13.85±3.96)較對(duì)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(shí)驗(yàn)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(duì)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(shí)驗(yàn)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(xì)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(jī)制與能夠抑制FGFR的激活下調(diào)vimentin、MMP14和MMP2的表達(dá)有關(guān)。提示成纖維細(xì)胞生長(zhǎng)因子受體是治療膠質(zhì)瘤的一個(gè)有效靶點(diǎn)。
[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(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R739.41

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相關(guān)期刊論文 前1條

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本文編號(hào)：1503254