本文選題：Rb94基因　切入點(diǎn)：腺病毒表達(dá)載體　出處：《中國協(xié)和醫(yī)科大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

【摘要】： 視網(wǎng)膜母細(xì)胞瘤(Retinoblastoma, Rb)基因是最早克隆出來的人類腫瘤抑制基因,Rb基因的缺失和突變將引起細(xì)胞無限生長和繁殖,進(jìn)而導(dǎo)致腫瘤的發(fā)生。到目前為止,以Rb為靶基因的抗腫瘤基因治療的研究報(bào)導(dǎo)很多。近期研究發(fā)現(xiàn),N端缺失的Rb蛋白—PRb94(野生型全長PRb110的NH2末端缺失112個(gè)氨基酸殘基)對頭頸部癌、膀胱癌、前列腺癌等細(xì)胞系具有比PRb110更強(qiáng)的腫瘤抑制效應(yīng)。放射治療是臨床上腫瘤治療的經(jīng)典方法之一,近年有人將放療與病毒介導(dǎo)的基因治療聯(lián)合使用,有明顯的相互增強(qiáng)的作用。本實(shí)驗(yàn)的目的是探討外源性Rb94基因聯(lián)合放射對食管癌細(xì)胞系K150生長的抑制作用,為臨床上進(jìn)行Rb94基因-放射治療腫瘤奠定實(shí)驗(yàn)基礎(chǔ)。 本研究將重組腺病毒Ad-Rb94于體外轉(zhuǎn)染K150細(xì)胞,轉(zhuǎn)染后進(jìn)行6Gy137Csγ射線照射,分為空白對照組、Ad-LacZ對照組、Ad-Rb94組、照射組和Ad-Rb94聯(lián)合照射組,觀察K150細(xì)胞的生長曲線、細(xì)胞周期、細(xì)胞凋亡和Rb94基因表達(dá)的變化。并觀察荷人體腫瘤裸鼠在Ad-Rb94單獨(dú)作用及與輻射協(xié)同作用下腫瘤體積、相對腫瘤體積、瘤重、相對腫瘤增殖率以及腫瘤體積生長變化的時(shí)間曲線,評價(jià)其抗腫瘤活性。 從我們所得的實(shí)驗(yàn)結(jié)果可知,Ad-Rb94轉(zhuǎn)染K150細(xì)胞后,Ad-Rb94組、照射組和聯(lián)合照射組對K150細(xì)胞生長均具有抑制作用,聯(lián)合照射組的抑制效應(yīng)最強(qiáng),明顯高于Ad-Rb94組和照射組。聯(lián)合照射組K150細(xì)胞中Rb94基因表達(dá)量最多,分別是Ad-Rb94組和空白對照組的1.87倍和2.92倍。荷人體腫瘤裸鼠在Ad-Rb94單獨(dú)作用及輻射協(xié)同作用下腫瘤體積、相對腫瘤體積、瘤重均低于對照組,而且聯(lián)合照射組的腫瘤生長受抑制效應(yīng)最強(qiáng)。 結(jié)果說明單獨(dú)Rb94基因或照射均對K150細(xì)胞的生長具有一定的抑制作用,而Ad-Rb94聯(lián)合放射抑瘤效應(yīng)最明顯,有效地抑制腫瘤細(xì)胞的生長。
[Abstract]:Retinoblastoma (RB) gene is the earliest cloned human tumor suppressor gene. The deletion and mutation of RB gene will lead to unlimited cell growth and reproduction, and lead to tumorigenesis. There are many studies on antitumor gene therapy targeting RB gene. Recently, it has been found that RB protein-PRb94 (NH2 terminal deletion of wild-type full-length PRb110) is associated with head and neck cancer and bladder cancer. Prostate cancer and other cell lines have stronger tumor inhibitory effect than PRb110. Radiotherapy is one of the classical methods of clinical tumor therapy. In recent years, radiotherapy has been combined with virus-mediated gene therapy. The aim of this study was to investigate the inhibitory effect of exogenous Rb94 gene combined with radiation on the growth of esophageal carcinoma cell line K150, and to lay an experimental foundation for clinical application of Rb94 gene-radiotherapy. In this study, recombinant adenovirus Ad-Rb94 was transfected into K150 cells in vitro. After transfection, the cells were irradiated by 6Gy137Cs 緯 -ray. The cells were divided into control group (Ad-LacZ control group) and Ad-Rb94 group. The growth curve and cell cycle of K150 cells were observed. The changes of apoptosis and Rb94 gene expression in nude mice bearing human tumor were observed. The time curves of tumor volume, relative tumor volume, tumor weight, relative tumor proliferation rate and tumor volume growth were observed in nude mice bearing human tumor under the action of Ad-Rb94 alone and in coordination with radiation. To evaluate its anti-tumor activity. From our experimental results, we can see that the growth of K150 cells was inhibited by Ad-Rb94 after transfection with Ad-Rb94, and the growth of K150 cells was inhibited by the combined irradiation group and the combined irradiation group, and the inhibitory effect was the strongest in the combined irradiation group. The expression of Rb94 gene in K150 cells in combined irradiation group was 1.87 times and 2.92 times higher than that in Ad-Rb94 group and blank control group, respectively. The tumor weight was lower than that of the control group, and the tumor growth inhibition was strongest in the combined irradiation group. The results showed that Rb94 gene alone or irradiation could inhibit the growth of K150 cells to a certain extent, while Ad-Rb94 combined with radiation had the most obvious inhibitory effect on the growth of K150 cells.
【學(xué)位授予單位】：中國協(xié)和醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R739.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

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本文編號：1649896