本文選題：腦膜瘤 + 骨膜蛋白��； 參考：《南方醫(yī)科大學(xué)》2017年碩士論文

【摘要】：研究背景與目的腦膜瘤是起源于蛛網(wǎng)膜帽狀細(xì)胞的顱內(nèi)最常見(jiàn)的非神經(jīng)上皮性腫瘤。絕大多數(shù)腦膜瘤為良性腫瘤(WHO I級(jí)),但仍然有高達(dá)25%左右的腦膜瘤為非典型腦膜瘤(WHO II級(jí))和惡性腦膜瘤(WHO III級(jí))。惡性腦膜瘤的預(yù)后最差,中位生存期僅為1.5年,總體復(fù)發(fā)率高達(dá)80%,腫瘤的病理分級(jí)和侵襲是影響復(fù)發(fā)和預(yù)后的關(guān)鍵因素。因此,尋找新的有效治療方法成為了目前改善惡性腦膜瘤預(yù)后的首要目標(biāo)。在我們的前期研究中,發(fā)現(xiàn)骨膜蛋白(periostin,POSTN)在不同級(jí)別中的腦膜瘤間質(zhì)中呈不同程度的表達(dá),并且其表達(dá)量也隨著病理等級(jí)的增高而增高。骨膜蛋白作為一種細(xì)胞外基質(zhì)(extracellular matrix,ECM)蛋白,可與整合素受體相結(jié)合,參與腫瘤微環(huán)境的調(diào)控,促進(jìn)腫瘤的侵襲與增殖,導(dǎo)致腫瘤發(fā)生上皮間質(zhì)轉(zhuǎn)化(epithelial-mesenchymal transition,EMT)等惡性行為。但骨膜蛋白在腦膜瘤中的作用及分子機(jī)制尚未有研究報(bào)道。本項(xiàng)研究通過(guò)對(duì)骨膜蛋白在惡性腦膜瘤中的分子機(jī)制的研究,探討了促進(jìn)其侵襲和增殖的信號(hào)通路及關(guān)鍵蛋白,可為惡性腦膜瘤的綜合治療提供潛在的治療靶點(diǎn)及新的啟示。研究?jī)?nèi)容與方法1.惡性腦膜瘤原代細(xì)胞的培養(yǎng)與鑒定收集新鮮標(biāo)本進(jìn)行原代細(xì)胞培養(yǎng)并純化,觀察原代細(xì)胞形態(tài),測(cè)定增殖能力,免疫熒光鑒定細(xì)胞。2.骨膜蛋白對(duì)惡性腦膜瘤細(xì)胞功能及表型的研究不同級(jí)別的腦膜瘤組織進(jìn)行qRT-PCR和Western-blot檢測(cè)POSTN的表達(dá)量,免疫組化法觀察POSTN在不同級(jí)別腦膜瘤中的表達(dá)程度及在惡性腦膜瘤中的表達(dá)部位,Spearman相關(guān)性檢測(cè)法檢測(cè)POSTN與惡行腦膜瘤侵襲的相關(guān)性,Western-blot檢測(cè)分離純化后的惡性腦膜瘤細(xì)胞與腫瘤相關(guān)成纖維細(xì)胞中POSTN的表達(dá)量。3.骨膜蛋白影響惡性腦膜瘤惡性表型的機(jī)制研究原代培養(yǎng)的惡性腦膜瘤細(xì)胞分別轉(zhuǎn)染過(guò)表達(dá)POSTN病毒與空載病毒,Western-blot檢測(cè)兩組細(xì)胞EMT相關(guān)蛋白的變化情況以及Akt、pAkt的變化情況。研究結(jié)果1.原代培養(yǎng)的腦膜瘤細(xì)胞經(jīng)鑒定EMA及Vimentin表達(dá)陽(yáng)性,惡性腦膜瘤原代細(xì)胞增殖能力強(qiáng)于良性腦膜瘤細(xì)胞,良性腦膜瘤細(xì)胞中β-半乳糖苷酶染色陽(yáng)性率更高。2.惡性腦膜瘤中POSTN蛋白水平和mRNA水平表達(dá)都高于良性腦膜瘤(P0.01),POSTN與惡性腦膜瘤侵襲顯著相關(guān)(r=0.738,p0.001),POSTN表達(dá)于惡性腦膜瘤的間質(zhì)中,原代培養(yǎng)中的成纖維細(xì)胞POSTN含量高于腦膜瘤細(xì)胞(P0.01)。3.惡性腦膜瘤細(xì)胞過(guò)表達(dá)POSTN后E-ca明顯下調(diào),N-ca、p-Akt、Vimentin、Snail明顯上調(diào)(P0.001);增殖能力及侵襲能力增強(qiáng)(P0.001)。研究結(jié)論惡性腦膜瘤與良性腦膜瘤相比,具備更強(qiáng)的增殖能力,也更具有侵襲性,這可能是導(dǎo)致高級(jí)別腦膜瘤預(yù)后更差的原因。由高級(jí)別腦膜瘤中的腫瘤相關(guān)成纖維細(xì)胞分泌的細(xì)胞外基質(zhì)蛋白——骨膜蛋白,可能是調(diào)控高級(jí)別腦膜瘤惡性表型的一個(gè)關(guān)鍵蛋白。骨膜蛋白可促進(jìn)Akt的磷酸化使其相關(guān)通路激活,影響高級(jí)別腦膜瘤的EMT過(guò)程從而促進(jìn)腫瘤的侵襲。因此,針對(duì)這一調(diào)控機(jī)制的靶向治療,或許可以對(duì)高級(jí)別腦膜瘤的綜合治療提供新的靶點(diǎn),改善惡性腦膜瘤患者的預(yù)后。
[Abstract]:Background and objective meningiomas are the most common non neuroepithelial tumors of the intracranial arachnoid cap cells. Most meningiomas are benign tumors (WHO I grade), but there are still about 25% of meningiomas as atypical meningiomas (WHO II) and malignant meningiomas (WHO III). The worst prognosis of malignant meningiomas is the middle stage of malignant meningioma. The survival time is only 1.5 years and the overall recurrence rate is up to 80%. The pathological classification and invasion of the tumor are the key factors affecting the recurrence and prognosis. Therefore, finding new effective treatments has become the primary goal of improving the prognosis of malignant meningioma. In our previous study, we found the meninges of Periostin (POSTN) in different levels. The expression of the tumor is expressed in varying degrees and its expression increases with the increase of the pathological grade. As an extracellular matrix (extracellular matrix, ECM) protein, periosteum protein can be combined with integrin receptor, participate in the regulation of tumor microenvironment, promote tumor invasion and proliferation, and lead to epithelial mesenchymal transformation of tumor (EP Ithelial-mesenchymal transition, EMT) and other malignant behaviors. However, the role and molecular mechanism of periosteum protein in meningioma have not been reported. This study has explored the signal pathways and key proteins that promote the invasion and proliferation of the periosteum protein in the malignant meningioma, and can be used as a synthesis of the malignant meningioma. Combination therapy provides potential therapeutic targets and new implications. Research contents and methods 1. primary cells of malignant meningioma are cultured and identified to collect fresh specimens for primary cell culture and purification, observe the morphology of primary cells, determine the proliferation ability, and immunofluorescence identification of.2. periosteum for the function and phenotype of malignant meningioma cells. The expression of POSTN was detected by qRT-PCR and Western-blot in different meningioma tissues. The expression level of POSTN in different meningiomas and the expression of POSTN in different meningiomas were observed by immunohistochemistry. The correlation between POSTN and the invasion of malignant meningioma was detected by Spearman correlation detection. Western-blot was used to detect the separation and purity of the meningioma. The expression of POSTN in malignant meningioma cells and tumor related fibroblasts.3. effect of periosteum protein on malignant phenotype of malignant meningioma; the primary cultured malignant meningioma cells were transfected with POSTN virus and airborne virus respectively. The changes of EMT related proteins in two groups of cells were detected by Western-blot and Akt, pA KT changes. Results 1. primary cultured meningioma cells were identified as positive for EMA and Vimentin. The primary cell proliferation of malignant meningioma was stronger than that of benign meningioma cells. The positive rate of beta galactosidase staining in benign meningioma cells was higher than that in.2. malignant meningioma. The expression of POSTN protein and mRNA was higher than that of benign meningioma. P0.01, POSTN was significantly associated with malignant meningioma (r=0.738, p0.001). POSTN was expressed in the stroma of malignant meningioma. The content of POSTN in primary cultured fibroblasts was higher than that of meningioma cells (P0.01).3. malignant meningioma cells. Colonization and invasiveness enhanced (P0.001). Conclusions malignant meningiomas are more proliferative and aggressive than benign meningiomas, which may be the cause of worse prognosis in the high grade meningioma. The extracellular matrix protein secreted by the tumor related fibroblast in the high level meningioma, the periosteum egg White, may be a key protein in the regulation of the malignant phenotype of high grade meningiomas. Periosteum can promote the phosphorylation of Akt to activate its pathway, affect the EMT process of the high level meningioma and promote the invasion of the tumor. Therefore, targeted therapy for this regulatory mechanism, or permits for the comprehensive treatment of high level meningiomas, may be provided. The new target is to improve the prognosis of patients with malignant meningioma.

【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)】：R739.45

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