【摘要】：目的:原發(fā)的成人腦腫瘤中膠質(zhì)瘤是最常見(jiàn)的類(lèi)型。它侵襲性強(qiáng),與周?chē)M織界限不清晰。因此,常規(guī)的治療手段如手術(shù)治療和放射治療對(duì)它作用都很有限。不但如此,膠質(zhì)瘤的耐藥性也導(dǎo)致其化學(xué)治療收獲甚微。因此,膠質(zhì)瘤患者需要更有效的治療方案,而這些治療方案的建立需要我們找到新的分子靶點(diǎn)。核黃素轉(zhuǎn)運(yùn)基因2(RFT2)是SLC52家族的一員,它在核黃素的代謝中起著重要的作用。最近,一些研究已經(jīng)證實(shí),RFT2在許多人類(lèi)腫瘤(食管鱗狀細(xì)胞癌、胃癌、宮頸癌)的發(fā)生發(fā)展中十分重要。本研究旨在探究RFT2在人類(lèi)膠質(zhì)瘤中的表達(dá)情況及其對(duì)膠質(zhì)瘤細(xì)胞生物學(xué)特性的作用,進(jìn)而為膠質(zhì)瘤的分子治療提供新的靶點(diǎn)。方法:為了檢測(cè)RFT2在正常腦組織和人類(lèi)腦膠質(zhì)瘤組織中的表達(dá)水平,我們對(duì)120例不同級(jí)別的膠質(zhì)瘤組織、24例正常腦組織進(jìn)行了免疫組織化學(xué)染色。利用RFT2-siRNA-3抑制膠質(zhì)瘤細(xì)胞系LN308和LN229中RFT2的表達(dá)。使用MTT、平板克隆形成實(shí)驗(yàn)、流式細(xì)胞術(shù)、Transwell實(shí)驗(yàn)、Western blot和裸鼠皮下荷瘤模型從體內(nèi)和體外兩方面來(lái)進(jìn)行膠質(zhì)瘤細(xì)胞增殖、遷移、侵襲等表型的實(shí)驗(yàn)評(píng)估。結(jié)果:1、與正常腦組織相比RFT2在膠質(zhì)瘤組織中是高表達(dá)的。而且,膠質(zhì)瘤中RFT2的表達(dá)水平與WHO分級(jí)呈正相關(guān)。2、與對(duì)照組和無(wú)義序列轉(zhuǎn)染組相比,敲低LN308和LN229細(xì)胞RFT2的表達(dá)后細(xì)胞增殖被明顯抑制(P0.01),細(xì)胞集落形成率明顯下降(P0.01)。敲低RFT2可有效抑制移植瘤的體內(nèi)生長(zhǎng)(P0.001)。3、敲低RFT2后LN308和LN229細(xì)胞阻滯在G1/G0期(P0.05)。在LN308和LN229細(xì)胞中敲低RFT2可以明顯升高其凋亡細(xì)胞的比例(P0.05)。各組中細(xì)胞周期和凋亡相關(guān)調(diào)節(jié)蛋白(p21、p27、Bcl-2、Bax)的表達(dá)都明顯改變。4、敲低RFT2后LN308和LN229細(xì)胞的遷移和侵襲能力受到抑制,其相關(guān)調(diào)節(jié)蛋白MMP-2和MMP-9的表達(dá)水平也相應(yīng)下降。結(jié)論:本研究發(fā)現(xiàn),膠質(zhì)瘤組織中RFT2呈高表達(dá),它的表達(dá)水平與膠質(zhì)瘤WHO分級(jí)呈正相關(guān)。敲低RFT2促進(jìn)了LN308和LN229細(xì)胞的細(xì)胞周期阻滯、細(xì)胞凋亡,抑制了細(xì)胞增殖、遷移和侵襲的能力。不但如此,裸鼠荷瘤模型也進(jìn)一步更好地證明了敲低RFT2后膠質(zhì)瘤細(xì)胞的增殖受到抑制。這些結(jié)果提示,RFT2可以成為膠質(zhì)瘤分子治療的新靶點(diǎn)。
[Abstract]:Objective: gliomas are the most common type of primary adult brain tumors. It is aggressive, and its boundaries with surrounding tissues are not clear. As a result, conventional treatments such as surgery and radiotherapy have limited effects on it. Moreover, the drug resistance of gliomas leads to little success in chemotherapy. Therefore, glioma patients need more effective treatment protocols, and the establishment of these protocols requires us to find new molecular targets. Riboflavin transporter gene 2 (RFT2) is a member of SLC52 family and plays an important role in riboflavin metabolism. Recently, some studies have confirmed that RFT2 plays an important role in the development of many human tumors (esophageal squamous cell carcinoma, gastric cancer, cervical cancer). The purpose of this study was to investigate the expression of RFT2 in human glioma and its effect on the biological characteristics of glioma cells, and to provide a new target for the molecular therapy of glioma. Methods: in order to detect the expression of RFT2 in normal brain tissues and human glioma tissues, immunohistochemical staining was performed in 120 gliomas of different grades and 24 normal brain tissues. The expression of RFT2 in glioma cell line LN308 and LN229 was inhibited by RFT2-siRNA-3. The phenotypes of glioma cell proliferation, migration and invasion were evaluated in vivo and in vitro by MTT, plate clone formation assay, flow cytometry, Transwell assay, Western blot and nude mice subcutaneous tumor model. Results: 1. The expression of RFT2 in glioma was higher than that in normal brain tissue. Moreover, the expression of RFT2 in gliomas was positively correlated with the WHO grade. 2Compared with the control group and the nonsense sequence transfection group, the proliferation of LN308 and LN229 cells was significantly inhibited after the expression of RFT2 was lowered (P0.01). The colony formation rate decreased significantly (P0.01). Knockout of RFT2 could effectively inhibit the growth of transplanted tumor in vivo (P0.001). 3. LN308 and LN229 cells were blocked in G1/G0 phase after knock down RFT2 (P0.05). Knockout of RFT2 in LN308 and LN229 cells significantly increased the percentage of apoptotic cells (P0.05). The cell cycle and the expression of apoptosis-related regulatory protein (p21, p27, Bcl-2, Bax) were significantly changed in each group. 4. The migration and invasion of LN308 and LN229 cells were inhibited after knockout with RFT2. The expression levels of MMP-2 and MMP-9 were also decreased. Conclusion: high expression of RFT2 was found in glioma tissues, and its expression level was positively correlated with WHO grade of gliomas. Knockout of RFT2 promoted cell cycle arrest, apoptosis and cell proliferation, migration and invasion of LN308 and LN229 cells. In addition, the nude mice tumor-bearing model further demonstrated that the proliferation of glioma cells was inhibited after knockout of RFT2. These results suggest that RFT2 may be a new target for glioma molecular therapy.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R739.41

【參考文獻(xiàn)】

相關(guān)期刊論文 前8條

1 楊如會(huì);宋羽;吳松泉;沈祥春;;干擾核黃素代謝對(duì)細(xì)胞卵巢癌HO8910細(xì)胞順鉑敏感性的影響（英文）[J];中國(guó)藥理學(xué)與毒理學(xué)雜志;2016年08期

2 張曉娟;石勁松;張玲染;徐小溪;侯素平;張清泉;;食管鱗狀細(xì)胞癌組織中PLCE1和RFT2的表達(dá)及臨床意義[J];臨床與實(shí)驗(yàn)病理學(xué)雜志;2015年12期

3 穆艾太爾·麥提努日;瑪依努爾·艾力;;核黃素與胃癌[J];國(guó)際腫瘤學(xué)雜志;2015年09期

4 黃靜;劉利敏;楊建學(xué);劉玉;李洪雷;王建坡;許麗艷;陳繼理;李恩民;周福有;王立東;;賁門(mén)癌和其癌旁組織核黃素轉(zhuǎn)運(yùn)基因全基因外顯子測(cè)序[J];鄭州大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2012年05期

5 Maynur Eli;Maimaitiaili Wumar;Batur Mamtimin;Ilyar Sheyhidin;Ayshamgul Hasim;;Decreased blood riboflavin levels are correlated with defective expression of RFT2 gene in gastric cancer[J];World Journal of Gastroenterology;2012年24期

6 李紀(jì)賓;鄒小農(nóng);王華余;陶德明;喬友林;顧元?jiǎng)P;鄭素芳;;鹽亭縣核黃素強(qiáng)化鹽干預(yù)試驗(yàn)人群食管癌前病變與轉(zhuǎn)歸的研究[J];中華腫瘤防治雜志;2009年05期

7 呂全軍;王華麗;胡同宇;朱明君;王旗;陸維權(quán);薛樂(lè)勛;劉志才;連士勇;李變?cè)?程蘭平;魏建榮;;林州市食管癌高發(fā)區(qū)人群維生素營(yíng)養(yǎng)狀況的動(dòng)態(tài)變化研究[J];衛(wèi)生研究;2007年06期

8 鄭素芳;核黃素與癌[J];中國(guó)腫瘤;1994年06期

相關(guān)會(huì)議論文 前1條

1 鄭素芳;林培中;喬長(zhǎng)虹;張金生;宋林華;劉新伏;丁鎮(zhèn)偉;王繼信;馮華進(jìn);;核黃素缺乏對(duì)甲基芐基亞硝胺誘發(fā)大鼠食管癌的影響[A];中國(guó)營(yíng)養(yǎng)學(xué)會(huì)第七屆全國(guó)營(yíng)養(yǎng)學(xué)術(shù)會(huì)議論文摘要匯編[C];1996年

，

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