人Rb94基因联合γ射线照射对K150细胞生长影响的体内、外研究

发布时间：2018-03-22 18:41

本文选题：Rb94基因　切入点：腺病毒表达载体　出处：《中国协和医科大学》2010年硕士论文　论文类型：学位论文

【摘要】： 视网膜母细胞瘤(Retinoblastoma, Rb)基因是最早克隆出来的人类肿瘤抑制基因,Rb基因的缺失和突变将引起细胞无限生长和繁殖,进而导致肿瘤的发生。到目前为止,以Rb为靶基因的抗肿瘤基因治疗的研究报导很多。近期研究发现,N端缺失的Rb蛋白—PRb94(野生型全长PRb110的NH2末端缺失112个氨基酸残基)对头颈部癌、膀胱癌、前列腺癌等细胞系具有比PRb110更强的肿瘤抑制效应。放射治疗是临床上肿瘤治疗的经典方法之一,近年有人将放疗与病毒介导的基因治疗联合使用,有明显的相互增强的作用。本实验的目的是探讨外源性Rb94基因联合放射对食管癌细胞系K150生长的抑制作用,为临床上进行Rb94基因-放射治疗肿瘤奠定实验基础。本研究将重组腺病毒Ad-Rb94于体外转染K150细胞,转染后进行6Gy137Csγ射线照射,分为空白对照组、Ad-LacZ对照组、Ad-Rb94组、照射组和Ad-Rb94联合照射组,观察K150细胞的生长曲线、细胞周期、细胞凋亡和Rb94基因表达的变化。并观察荷人体肿瘤裸鼠在Ad-Rb94单独作用及与辐射协同作用下肿瘤体积、相对肿瘤体积、瘤重、相对肿瘤增殖率以及肿瘤体积生长变化的时间曲线,评价其抗肿瘤活性。从我们所得的实验结果可知,Ad-Rb94转染K150细胞后,Ad-Rb94组、照射组和联合照射组对K150细胞生长均具有抑制作用,联合照射组的抑制效应最强,明显高于Ad-Rb94组和照射组。联合照射组K150细胞中Rb94基因表达量最多,分别是Ad-Rb94组和空白对照组的1.87倍和2.92倍。荷人体肿瘤裸鼠在Ad-Rb94单独作用及辐射协同作用下肿瘤体积、相对肿瘤体积、瘤重均低于对照组,而且联合照射组的肿瘤生长受抑制效应最强。结果说明单独Rb94基因或照射均对K150细胞的生长具有一定的抑制作用,而Ad-Rb94联合放射抑瘤效应最明显,有效地抑制肿瘤细胞的生长。
[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 纬 -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.
【学位授予单位】：中国协和医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.7

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