本文選題：格列本脲　切入點(diǎn)：膠質(zhì)母細(xì)胞瘤　出處：《重慶醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的腫瘤是嚴(yán)重危害人類健康的四大疾病(心臟病、腫瘤、腦血管病、阿爾茨海默病)之一,由細(xì)胞異常分化、增殖形成,化療作為治療腫瘤的主要手段,常常引起眾多副反應(yīng),如:免疫功能極其低下、骨髓功能受到抑制等,并且在化療過程中,患者還出現(xiàn)對藥物的耐藥性,這更增加了醫(yī)療上治療的難度。星形膠質(zhì)瘤是腫瘤中常見類型,按照其病理學(xué)特點(diǎn),世界衛(wèi)生組織將其分為四個病理級別,其中膠質(zhì)母細(xì)胞瘤是Ⅳ級,屬于高惡性膠質(zhì)瘤,侵襲性較強(qiáng),發(fā)病率極高,且術(shù)后容易復(fù)發(fā),致死率高,目前臨床治療效果不盡如意。因此,研究膠質(zhì)瘤的發(fā)病機(jī)制并探尋新的治療手段迫在眉睫。方法:本課題使用藥物格列本脲作用膠質(zhì)瘤細(xì)胞U251和U87,初步探索其對U251和U87的影響及機(jī)制研究。應(yīng)用CCK8、劃痕法、Transwell遷移與侵襲、Hocheset染色、流式細(xì)胞檢測、p H探針、蛋白免疫印跡(Western Blot)等多種分子生物學(xué)和電生理學(xué)實(shí)驗(yàn)技術(shù),檢測格列本脲對膠質(zhì)瘤U251和U87細(xì)胞增殖生長的影響及機(jī)制。體外培養(yǎng)人膠質(zhì)瘤U251和U87細(xì)胞,隨機(jī)分為實(shí)驗(yàn)組及對照組。利用不同濃度的格列本脲作用于體外培養(yǎng)的U251和U87細(xì)胞,共培養(yǎng)48 h,以未加入格列本脲的細(xì)胞組別為對照組,CCK8測定細(xì)胞的增殖能力。劃痕法及Transwell觀察細(xì)胞遷移和侵襲。Hocheset染色及流式細(xì)胞術(shù)檢測細(xì)胞凋亡變化。p H探針觀察細(xì)胞內(nèi)酸堿度改變。Western Blot檢測細(xì)胞中Kir4.1和MCT1的表達(dá)水平結(jié)果CCK8顯示在一定濃度范圍內(nèi),格列本脲可抑制細(xì)胞增殖(P0.05),并呈濃度依賴性(P0.05);劃痕法顯示格列本脲抑制細(xì)胞遷移(P0.05),Transwell法顯示實(shí)驗(yàn)組遷移與侵襲細(xì)胞數(shù)量明顯減少(P0.05),且隨著藥物濃度增高細(xì)胞數(shù)量逐漸減少(P0.05);Hochest法及流式檢測顯示實(shí)驗(yàn)組出現(xiàn)明顯凋亡特征,且隨著格列本脲濃度的升高,凋亡現(xiàn)象越明顯(P0.05);熒光探針顯示隨著藥物濃度的增加,熒光強(qiáng)度逐漸減弱(P0.05),提示藥物濃度越高,胞內(nèi)p H逐漸降低;Western Blot結(jié)果表明實(shí)驗(yàn)組細(xì)胞的Kir4.1和MCT1蛋白含量明顯降低(P0.05),且隨藥物濃度的增高而下降(P0.05)。結(jié)論1格列本脲能夠抑制膠質(zhì)母細(xì)胞瘤增殖生長;2格列本脲通過影響Kir4.1和MCT1的表達(dá),參與調(diào)節(jié)內(nèi)環(huán)境酸堿度,發(fā)揮增殖抑制作用。
[Abstract]:Objective tumor is one of the four major diseases (heart disease, tumor, cerebrovascular disease, Alzheimer's disease) which is seriously harmful to human health. It often causes many side effects, such as the extremely low immune function, the suppression of bone marrow function, and the drug resistance of patients during chemotherapy. This adds to the difficulty of medical treatment. Astrocytoma is a common type of tumor. According to its pathological characteristics, the World Health Organization divides glioblastoma into four pathological grades, among which glioblastoma is grade 鈪，

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