【摘要】：目的:不同膠質(zhì)瘤細(xì)胞系對(duì)全反式維甲酸(ATRA)的反應(yīng)不同,可能與脂肪酸結(jié)合5(FABP5)結(jié)合ATRA后激活促細(xì)胞增殖通路有關(guān)。此外,有研究報(bào)道,FABP5與腫瘤的發(fā)生、發(fā)展以及轉(zhuǎn)移相關(guān),而FABP5在膠質(zhì)瘤中的功能尚不清楚。本研究旨在探討FABP5蛋白在不同級(jí)別膠質(zhì)瘤中的表達(dá)、功能及其與ATRA敏感性的關(guān)系,為后續(xù)進(jìn)一步研究FABP5相關(guān)的膠質(zhì)瘤靶向治療藥物提供新思路。方法:1.免疫組化檢測(cè)23例腫瘤旁正常腦組織以及137例不同級(jí)別膠質(zhì)瘤中FABP5的表達(dá),統(tǒng)計(jì)分析FABP5的表達(dá)與臨床病理參數(shù)之間的關(guān)系;并根據(jù)臨床隨訪資料,繪制生存曲線。2.體外培養(yǎng)人腦膠質(zhì)瘤細(xì)胞株A172、T98G和U251,利用蛋白免疫印跡法(Western blot)檢測(cè)3種膠質(zhì)瘤細(xì)胞系中FABP5蛋白表達(dá)水平;采用si RNA技術(shù)構(gòu)建相關(guān)細(xì)胞模型,下調(diào)A172、T98G細(xì)胞中FABP5蛋白表達(dá)水平,利用噻唑藍(lán)(MTT)比色法、劃痕實(shí)驗(yàn)和Transwell法分別檢測(cè)細(xì)胞增殖活力、遷移及侵襲變化。3.不同濃度ATRA(0,1,5,10mM)處理3種細(xì)胞,進(jìn)行蘇木素-伊紅(HE)染色,觀察用藥前后膠質(zhì)瘤細(xì)胞形態(tài)的變化;利用MTT法檢測(cè)ATRA對(duì)膠質(zhì)瘤細(xì)胞增殖活力的影響;利用Western blot和免疫細(xì)胞化學(xué)(ICC)法檢測(cè)膠質(zhì)瘤細(xì)胞用藥前后FABP5蛋白表達(dá)變化。結(jié)果:1.FABP5在膠質(zhì)瘤中的表達(dá)明顯高于正常腦組織(P0.05),且與膠質(zhì)瘤的組織學(xué)分級(jí)、預(yù)后相關(guān)(P均0.05)。2.FABP5在U251、A172、T98G細(xì)胞系中均有表達(dá),表現(xiàn)出依次遞增。與對(duì)照組相比較si RNA下調(diào)FABP5后能明顯降低A172、T98G細(xì)胞的增殖活力,遷移和侵襲能力顯著下調(diào)。3.用藥前后三種細(xì)胞形態(tài)均無(wú)明顯差異;MTT檢測(cè)結(jié)果顯示僅在48h時(shí)10mM ATRA對(duì)A172細(xì)胞有明顯抑制細(xì)胞生長(zhǎng)作用,不同濃度ATRA對(duì)A172、T98G細(xì)胞及1,5mM ATRA對(duì)A172均無(wú)明顯抑制作用;用藥前后FABP5蛋白總量及細(xì)胞亞分布均無(wú)明顯改變。結(jié)論:FABP5可以促進(jìn)膠質(zhì)瘤細(xì)胞的增殖、遷移與侵襲能力,并與膠質(zhì)瘤患者預(yù)后相關(guān)。膠質(zhì)瘤細(xì)胞系對(duì)ATRA欠敏感,與FABP5蛋白的表達(dá)無(wú)明顯相關(guān)性。
[Abstract]:AIM: Different glioma cell lines respond differently to all-trans retinoic acid (ATRA) and may be related to activation of the cell proliferation pathway after fatty acid binding 5 (FABP5) binding to ATRA. In addition, some studies have reported that FABP5 is associated with tumorigenesis, development and metastasis, while the function of FABP5 in glioma remains unclear. The expression and function of FABP5 in different grades of gliomas and the relationship between FABP5 and ATRA sensitivity provide new ideas for further study of FABP5-related targeted therapy drugs for gliomas. Methods: 1. Immunohistochemistry was used to detect the expression of FABP5 in 23 normal brain tissues adjacent to tumors and 137 gliomas of different grades. The expression of FABP5 protein in human glioma cell lines A172, T98G and U251 was detected by Western blot, and the related cell models were constructed by Si RNA technique to down-regulate the expression of FAB in A172 and T98G cells. P5 protein expression was detected by MTT colorimetry, scratch test and Transwell method respectively. 3. Three kinds of cells were treated with different concentrations of ATRA (0,1,5,10 mM) and stained with hematoxylin-eosin (HE) to observe the morphological changes of glioma cells before and after treatment. Results: 1. The expression of FABP5 in gliomas was significantly higher than that in normal brain tissues (P 0.05), and correlated with histological grade and prognosis of gliomas (P 0.05). 2. FABP5 was found in U251, A172, T98G cell lines. Compared with the control group, the proliferation activity, migration and invasion ability of A172 and T98G cells were significantly decreased after down-regulation of FABP5 by Si RNA. There was no significant inhibitory effect of ATRA on A172, T98G cells and 1,5mM ATRA, and no significant change in total FABP5 protein and cell subdistribution before and after treatment. Conclusion: FABP5 can promote the proliferation, migration and invasion of glioma cells, and is related to the prognosis of glioma patients. No significant correlation was found.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R739.41

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