發(fā)布時間：2019-01-11 13:10

【摘要】：研究目的:1.研究GRM5(代謝性谷氨酸受體5)在膠質(zhì)瘤表達(dá)與膠質(zhì)瘤病理級別的相關(guān)性;2.研究GRM5對膠質(zhì)瘤U251細(xì)胞的遷移及侵襲能力的影響。研究方法:1.為了研究膠質(zhì)瘤組織中GRM5表達(dá)情況,我們收集了本院神經(jīng)外科13例膠質(zhì)瘤標(biāo)本,所有標(biāo)本均依據(jù)2007年世界衛(wèi)生組織(World Health Origanization,WHO)中樞神經(jīng)系統(tǒng)腫瘤分類標(biāo)準(zhǔn)進(jìn)行病理診斷并分級。應(yīng)用免疫組化的方法檢測膠質(zhì)瘤標(biāo)本中GRM5的表達(dá)情況。2.為了研究GRM5對膠質(zhì)瘤U251細(xì)胞的遷移能力的影響,我們設(shè)計合成了2條特異性針對人GRM5基因的小分子干擾RNA(GRM5-siRNA-1247,GRM5-siRNA-1587),及對照RNA(GRM5-siRNA-NC);轉(zhuǎn)染U251細(xì)胞株48h后用Western blot(蛋白質(zhì)印跡法)檢測GRM5的表達(dá)情況;3.用轉(zhuǎn)染后的U251細(xì)胞做劃痕實驗和Tanswell實驗,驗證轉(zhuǎn)染小分子RNA(GRM5-siRNA-1247,GRM5-siRNA-1587),干擾GRM5表達(dá)后U251細(xì)胞的遷移和侵襲能力的影響。實驗結(jié)果:1.免疫組化結(jié)果顯示GRM5在人低級別和高級別膠質(zhì)瘤中均有表達(dá),且隨腫瘤級別增高,表達(dá)量也增高,結(jié)果具有統(tǒng)計學(xué)意義(P0.01)。2.Western Blot實驗結(jié)果顯示U251細(xì)胞轉(zhuǎn)染小分子干擾RNA(GRM5-siRNA-1247)后GRM5表達(dá)量明顯降低,差異具有統(tǒng)計學(xué)意義(t=3.020,P0.05)。3.劃痕實驗結(jié)果顯示U251細(xì)胞轉(zhuǎn)染小分子干擾RNA(GRM5-siRNA-1247)后,細(xì)胞劃痕傷口愈合率明顯低于對照組,差異具有統(tǒng)計學(xué)意義(24h t=6.751,P0.01;48h t=8.668 P0.01)。4.Tanswell實驗結(jié)果顯示U251細(xì)胞轉(zhuǎn)染小分子干擾RNA(GRM5-siRNA-1247)后,細(xì)胞侵襲能力明顯低于對照組,差異具有統(tǒng)計學(xué)意義(t=20.726,P0.01)。結(jié)論:1.GRM5在各級別膠質(zhì)瘤中都有表達(dá),且表達(dá)量與膠質(zhì)瘤級別呈正相關(guān)。2.GRM5能促進(jìn)膠質(zhì)瘤U251細(xì)胞遷移及侵襲的能力。
[Abstract]:Objective: 1. To study the correlation between the expression of GRM5 (metabolic glutamate receptor 5) in gliomas and the pathological grade of gliomas. 2. To study the effect of GRM5 on the migration and invasion of U251 glioma cells. Methods: 1. In order to study the expression of GRM5 in glioma tissues, we collected 13 glioma specimens from neurosurgery department of our hospital, all of which were based on WHO (World Health Origanization, in 2007. WHO) Central nervous system tumor classification criteria for pathological diagnosis and grading. Immunohistochemical method was used to detect the expression of GRM5 in glioma specimens. 2. In order to study the effect of GRM5 on the migration ability of U251 glioma cells, we designed and synthesized two small interfering RNA (GRM5-siRNA-1247,GRM5-siRNA-1587) specific to human GRM5 gene and control RNA (GRM5-siRNA-NC). The expression of GRM5 was detected by Western blot (Western blot after transfection of U251 cell line for 48 h. The effect of small molecule RNA (GRM5-siRNA-1247,GRM5-siRNA-1587) transfection on the migration and invasion of U251 cells after GRM5 expression was tested by scratch test and Tanswell assay. Experimental results: 1. Immunohistochemical results showed that GRM5 was expressed in both low-grade and high-grade gliomas, and the expression level increased with the increase of tumor grade. The results showed that the expression of GRM5 in U251 cells was significantly decreased after transfection of small molecule interfering RNA (GRM5-siRNA-1247) (P0.01), and the difference was statistically significant (p0.05). The results of scratch test showed that after transfection of small molecule interfering RNA (GRM5-siRNA-1247), the wound healing rate of U251 cells was significantly lower than that of the control group (24 hours after transfection with small molecule interference RNA (P0.01). The results of 4.Tanswell assay showed that the invasion ability of U251 cells transfected with small molecule interfering RNA (GRM5-siRNA-1247) was significantly lower than that of the control group (tr 20.726 P0.01). Conclusion: the expression of 1.GRM5 is positively correlated with the grade of glioma, and 2.GRM5 can promote the migration and invasion of U251 cells.
【學(xué)位授予單位】：廣州醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R739.41

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