本文選題：膠質(zhì)瘤 + 病理類型　； 參考：《中南大學(xué)》2014年博士論文

【摘要】：背景：高級(jí)別膠質(zhì)瘤是成人最常見的原發(fā)性顱內(nèi)惡性腫瘤。盡管近年來針對(duì)其手術(shù)、放療及化療等治療方案均取得了長足的進(jìn)展,患者的整體生存率依然較低,預(yù)后極差。世界衛(wèi)生組織(WHO)依據(jù)組織病理學(xué)將其分為4級(jí),其中膠質(zhì)母細(xì)胞瘤(GBM)是其中最常見、惡性程度最高的一類。GBM患者預(yù)后極差,其中位生存期為14.6個(gè)月,中位無進(jìn)展生存期為6.9個(gè)月,5年生存率為9.8%。 MicroRNA(MiRNA)是一種不參與蛋白質(zhì)編碼的小分子RNA,通過與靶基因mRNA3'-UTR序列互補(bǔ)的結(jié)合,抑制其表達(dá),實(shí)現(xiàn)對(duì)靶基因的負(fù)調(diào)控作用。最近的研究表明MiRNA的表達(dá)譜異常具有腫瘤特異性,并通過影響關(guān)鍵致癌／抑癌基因的表達(dá)參與腫瘤的發(fā)生、侵襲、抗凋亡等多種進(jìn)程。有證據(jù)表明單個(gè)MiRNA可影響多個(gè)靶基因的表達(dá)。目前,已有多個(gè)針對(duì)膠質(zhì)瘤與不同正常腦組織間MiRNA表達(dá)譜的差異的研究,均表現(xiàn)出了表達(dá)譜的差異,其在腫瘤進(jìn)展中的作用機(jī)制尚不完全明了。因此,發(fā)現(xiàn)并對(duì)在膠質(zhì)瘤中差異表達(dá)的MiRNA及其涉及的靶基因進(jìn)行深入研究有助于揭示其在細(xì)胞遷移、侵襲細(xì)胞骨架重排及血管形成中的作用,最終實(shí)現(xiàn)在診斷、治療中的應(yīng)用。 目的：針對(duì)不同病理類型膠質(zhì)瘤樣本MiRNA結(jié)果的篩查,篩選差異表達(dá)的MiRNA,用以發(fā)現(xiàn)新的膠質(zhì)瘤相關(guān)MiRNA并結(jié)合其表達(dá)模式推測其在膠質(zhì)瘤發(fā)生發(fā)展中的作用。深入研究MiR-125a-5p在GBM中的功能,并通過生物信息學(xué)的方式對(duì)其靶基因做出預(yù)測。 方法：分析膠質(zhì)瘤MiRNA數(shù)據(jù)庫,篩選出與病理類型相關(guān)的MiRNA。收集各種病理類型膠質(zhì)瘤標(biāo)本,并用qRT-PCR方法驗(yàn)證其表達(dá)量。向GBM細(xì)胞株轉(zhuǎn)染外源性MiR-125a-5p mimics,通過MTT法、劃痕實(shí)驗(yàn)與transwell侵襲實(shí)驗(yàn)分別檢測轉(zhuǎn)染前后細(xì)胞增殖、遷移、侵襲能力有無改變；應(yīng)用生物信息學(xué)方法,預(yù)測侵襲相關(guān)GMBMiRNAs可能的靶基因。 結(jié)果：(1)在膠質(zhì)瘤組織中發(fā)現(xiàn)42個(gè)在不同病理類型膠質(zhì)瘤中特異性表達(dá)的MiRNA,其中特異性下調(diào)的有7個(gè),特異性上調(diào)的有35個(gè)。(2)qRT-PCR實(shí)驗(yàn)結(jié)果與基因芯片結(jié)果相符,MiR-125a-5p在A/O等惡性程度較低的膠質(zhì)瘤中無明顯下調(diào),在AA/AOA/AO/GBM等惡性程度較高的膠質(zhì)瘤中下調(diào)明顯。其表達(dá)與MGMT的表達(dá)有相關(guān)性。(3)與對(duì)照組相比,轉(zhuǎn)染MiR-125a-5p mimics后96小時(shí)內(nèi)細(xì)胞增殖力的變化無統(tǒng)計(jì)學(xué)意義(P0.05)；(4)與陰性和空白對(duì)照組相比,轉(zhuǎn)染MiR-125a-5p mimics的GBM8401細(xì)胞侵襲及遷移能力明顯增強(qiáng)(P0.01);(5) MiR-125a-5p有多個(gè)靶基因,可能參與調(diào)節(jié)細(xì)胞生長、分化、凋亡、轉(zhuǎn)錄和信號(hào)轉(zhuǎn)導(dǎo)等多種過程相關(guān)。 結(jié)論：1.在膠質(zhì)瘤組織中發(fā)現(xiàn)42個(gè)在不同病理類型膠質(zhì)瘤中特異性表達(dá)的MiRNA,其中特異性下調(diào)的有7個(gè),特異性上調(diào)的有35個(gè)。 2. MiR-125a-5p在A/O等惡性程度較低的膠質(zhì)瘤中無明顯下調(diào),在AA/AOA/AO/GBM等惡性程度較高的膠質(zhì)瘤中下調(diào)明顯。其表達(dá)與MGMT的表達(dá)有相關(guān)性。 3.MiR-125a-5p的過表達(dá)可抑制GBM細(xì)胞的遷移及侵襲,對(duì)其生長增殖無明顯影響。
[Abstract]:Background : High - grade glioma is the most common primary intracranial malignant tumor in adults . Although in recent years , the overall survival rate of patients is low and the prognosis is extremely poor . Although in recent years , the overall survival rate of the patients is low and the prognosis is extremely poor . The World Health Organization ( WHO ) is divided into four grades according to the histopathology . The prognosis of the patients is extremely poor . Among them , the survival time is 14.6 months . Median progression - free survival is 6.9 months , and the 5 - year survival rate is 9.8 % .

MicroRNA ( MiRNA ) is a kind of small molecule RNA which is not involved in protein coding . It can inhibit the expression of target gene by binding to mRNA of mRNA3 ' - 3 ' of target gene . The recent research shows that the expression profile of MiRNA can influence the expression of multiple target genes .

Objective : To screen differentially expressed MiRNA for the screening of MiRNA results of different pathological types of glioma samples and to investigate the role of MiR - 125a - 5p in the development of glioma .

Methods : MiRNA of glioma was analyzed . MiRNA related to pathological type was selected . The expression of MiRNA was verified by qRT - PCR . The exogenous MiR - 125a - 5p - labeled cells were transfected with qRT - PCR . The proliferation , migration and invasion ability of transfected cells were detected by MTT assay , scratch test and transwell invasion assay .
Bioinformatic methods were used to predict the possible target genes of invasive related MRNA .

Results : ( 1 ) The specific expression of MiR - 125a - 5p and MiR - 125a - 5p in glioma tissues was significantly lower than that of control group , and the expression of MiR - 125a - 5p was significantly lower than that of control group ( P0.05 ) .
( 5 ) MiR - 125a - 5p has multiple target genes , which may be involved in regulating cell growth , differentiation , apoptosis , transcription and signal transduction .

Conclusion : 1 . MiRNA specifically expressed in 42 different pathological types of glioma was found in glioma tissues , among which there were 7 specific downregulation , 35 of which were up - regulated .

2 . MiR - 125a - 5p down - regulated in glioma with lower malignant degree , such as A / O and so on , down - regulated in high grade gliomas with higher malignant degree of AA / AOA / AO / . The expression of MiR - 125a - 5p was related to the expression of MGMT .

3 . The overexpression of MiR - 125a - 5p can inhibit the migration and invasion , and has no obvious effect on growth and proliferation .
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R739.41

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 ;Expression of miR-125b in the new,highly invasive glioma stem cell and progenitor cell line SU3[J];癌癥;2012年04期

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本文編號(hào)：2083744