【摘要】：背景和目的： 中樞神經(jīng)系統(tǒng)腫瘤中腦腫瘤占90%，其中膠質(zhì)瘤是最常見(jiàn)的腦腫瘤。膠質(zhì)瘤細(xì)胞具有高度的侵襲、遷移及增殖能力。然而，促進(jìn)腫瘤進(jìn)展和復(fù)發(fā)的分子機(jī)制仍不確定。因此，治療腫瘤的方法和藥物非常有限，且多數(shù)患者的中位生存率不足1年，膠質(zhì)母細(xì)胞瘤患者的臨床生存率則更低。因而，探索膠質(zhì)瘤細(xì)胞遷移及侵襲的分子機(jī)制是極有臨床意義的，可以促進(jìn)更有效的治療手段或藥物的研發(fā)。MicroRNAs是一類(lèi)18至25個(gè)核苷酸長(zhǎng)度的非編碼單鏈的小RNAs，通過(guò)調(diào)控特定的有活性的mRNA靶點(diǎn)，，可在惡性腫瘤的進(jìn)展過(guò)程起重要作用。超過(guò)半數(shù)的人類(lèi)基因由microRNA調(diào)控轉(zhuǎn)錄抑制和（或）mRNA降解來(lái)調(diào)控。MicroRNAs也參與腫瘤細(xì)胞的凋亡、增殖、遷移及侵襲的調(diào)控。研究表明，microRNA與腫瘤的發(fā)生有密切關(guān)系，并可分為致癌microRNA和抑癌microRNA。MicroRNA218為抑癌microRNA。近年來(lái)，MicroRNA218在一些腫瘤中已廣泛被研究，研究表明其在腦膠質(zhì)瘤細(xì)胞中顯著表達(dá)下調(diào)。然而，MicroRNA218對(duì)膠質(zhì)瘤細(xì)胞的遷移和侵襲的生物學(xué)影響知之甚少。本研究旨在探討microRNA218在腦膠質(zhì)瘤組織樣本中的表達(dá)水平與病理分級(jí)的相關(guān)性，以及microRNA218的生物學(xué)功能。 方法： （1）收集經(jīng)病理確診的不同級(jí)別膠質(zhì)瘤的新鮮組織樣本38例，用液氮罐轉(zhuǎn)運(yùn)至㧟80℃冰箱，用于總RNA的提取。 （2）應(yīng)用定量RT-PCR檢測(cè)miRNA218在各腦膠質(zhì)瘤組織樣本中的表達(dá)水平，并分析其與膠質(zhì)瘤病理分級(jí)的相關(guān)性。 （3）應(yīng)用脂質(zhì)體2000將pcDNA3.1-microRNA218質(zhì)粒轉(zhuǎn)染膠質(zhì)瘤細(xì)胞U87、U251。 （4）使用細(xì)胞計(jì)數(shù)試劑盒-8（CCK-8）細(xì)胞計(jì)數(shù)法檢測(cè)microRNA218對(duì)膠質(zhì)瘤細(xì)胞U87、U251存活率的影響。 （5）應(yīng)用Transwell侵襲實(shí)驗(yàn)檢測(cè)microRNA218對(duì)膠質(zhì)瘤細(xì)胞U87的侵襲能力的影響。 （6）應(yīng)用細(xì)胞劃痕實(shí)驗(yàn)測(cè)定microRNA218對(duì)膠質(zhì)瘤細(xì)胞U87的遷移能力的影響。 結(jié)果： (1)MicroRNA218在WHOI-II膠質(zhì)瘤患者中的表達(dá)水平明顯高于WHOIII-IV患者，且microRNA218的表達(dá)水平與腦膠質(zhì)瘤病理分級(jí)呈負(fù)相關(guān)。 (2)MicroRNA218抑制U87MG和U251MG細(xì)胞增殖能力。 (3)MicroRNA218體外抑制膠質(zhì)瘤細(xì)胞U87遷移與侵襲能力。結(jié)論： (1)MicroRNA218表達(dá)水平與病理分級(jí)呈負(fù)相關(guān)。 (2)MicroRNA218具有抑制膠質(zhì)瘤遷移與侵襲的作用。-
[Abstract]:Background and objective: brain tumors account for 90% of central nervous system tumors, of which glioma is the most common brain tumor. Glioma cells have high ability of invasion, migration and proliferation. However, the molecular mechanism for tumor progression and recurrence remains uncertain. Therefore, the treatment of tumors and drugs are very limited, and the median survival rate of most patients is less than 1 year, and the clinical survival rate of glioblastoma patients is even lower. Therefore, it is of great clinical significance to explore the molecular mechanism of migration and invasion of glioma cells. MicroRNAs are a class of non-coding single-stranded RNs with 18 to 25 nucleotides, which play an important role in the progression of malignant tumors by regulating specific active mRNA targets. More than half of human genes are regulated by microRNA to regulate transcription inhibition and / or mRNA degradation. MicroRNAs are also involved in the regulation of apoptosis, proliferation, migration and invasion of tumor cells. The results showed that microRNAs were closely related to tumorigenesis and could be divided into carcinogenic microRNA and tumor suppressor microRNA.MicroRNA218 as tumor suppressor microRNA. In recent years, microRNA218 has been widely studied in some tumors, and it has been shown that the expression of microRNA218 is down-regulated in glioma cells. However, little is known about the biological effects of MicroRNA218 on the migration and invasion of glioma cells. The purpose of this study was to investigate the correlation between the expression of microRNA218 in glioma tissues and the pathological grading, and the biological function of microRNA218. Methods: (1) 38 samples of different grade gliomas were collected and transported to 80 鈩

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