脂质体介导CK13基因对鼻咽癌放疗敏感性影响的动物实验研究

发布时间：2018-05-18 07:51

本文选题：CK13基因 + 鼻咽癌　；参考：《昆明医科大学》2012年硕士论文

【摘要】：目的：本文观察CK13基因对人鼻咽癌动物模型放射敏感性的影响。本课题是在前期研究基础上,利用HNE1细胞株和转染了目的基因的HNEl细胞株在Nu-Nu裸鼠腋下分别建立荷瘤动物模型,即NPC动物模型组和含目的基因的动物模型组,并在NPC动物模型组瘤体部位直接注射多价阳离子脂质体包裹表达CK13基因的质粒,对照组分别注射空载体质粒和磷酸盐缓冲液,然后所有组均给予2Gy直线加速器的局部肿瘤放射治疗(简称放疗),4天后重复肿瘤局部注射和放疗。观察肿瘤治疗前后大小变化,计算抑瘤率。最后处死小鼠,分离肿瘤组织。采用免疫组织学染色和RT-PCR的方法检测肿瘤组织CK13表达情况,HE染色比较各组移植瘤组织坏死程度。Western blotting进行CK13蛋白定量分析。探讨CK13基因与鼻咽癌放射敏感性的关系,以CK13基因为靶点进行相关研究,可为临床提高鼻咽癌放疗敏感性提供新的思路。方法：1.NPC动物模型及含目的基因NPC动物模型的建立。2.NPC动物模型局部注射质粒。3.放疗,并测量放疗前后肿瘤大小。4.处死小鼠,取标本做病检、免疫组化、RT-PCR、及Western Blotting。5.结果进行统计学分析。结果：在转染CK13基因的NPC动物模型组和注射有CK13基因的治疗组中CK13表达水平明显升高,CK13基因联合放疗治疗组肿瘤生长明显受限制。在第9天处死小鼠时,实验组(A1、A2、B1、B2组)的抑瘤率分别为44.6%,47.1%,45.9%,49.5%、空载体组(C1、C2组)抑瘤率为0,0.05%,各个实验组分别与空载体组比较,差异均有统计学差异(p0.05),各个实验组间相比较,差异无统计学意义(p0.05)。免疫组化检查实验组CK13阳性表达率较空载体组和对照组高,且差异有统计学意义。RT-PCR检测CK13基因显示：实验组CK13mRNA表达水平较空载体和对照组高。Western blotting示：实验组均可见明显CK13蛋白条带,而空载体组和对照组未见明显条带。结论：CK13基因可提高NPC对放射治疗的敏感性；CK13基因可能成为肿瘤治疗一个新的靶基因,为鼻咽癌的基因治疗提供一个新的途径。
[Abstract]:Objective: to observe the effect of CK13 gene on radiosensitivity of human nasopharyngeal carcinoma animal model. On the basis of previous studies, HNE1 cell line and HNEl cell line transfected with target gene were used to establish tumor-bearing animal models under the axillary of Nu-Nu nude mice, that is, NPC animal model group and target gene containing animal model group. The plasmid expressing CK13 gene was encapsulated by polyvalent cationic liposome directly into the tumor body of NPC model group, and the control group was injected with empty body mass and phosphate buffer respectively. Then all groups were given local tumor radiotherapy with 2Gy linear accelerator. The changes of tumor size before and after treatment were observed and the tumor inhibition rate was calculated. Finally, the mice were killed and tumor tissues were isolated. Immunohistochemical staining and RT-PCR were used to detect the expression of CK13 in tumor tissues. The degree of necrosis of transplanted tumor tissues in each group was compared with HE staining. Western blotting was used for quantitative analysis of CK13 protein. To explore the relationship between CK13 gene and radiosensitivity of nasopharyngeal carcinoma (NPC), using CK13 gene as the target to study the radiosensitivity of nasopharyngeal carcinoma (NPC) can provide a new way of thinking to improve the radiosensitivity of NPC. Methods 1. The animal model of NPC and the animal model containing target gene NPC. 2. Local injection of plasmid. 3. The tumor size was measured before and after radiotherapy. Mice were killed, specimens were collected for disease examination, immunohistochemical staining was performed with RT-PCR, and Western Blotting.5. Results Statistical analysis was carried out. Results: the expression of CK13 was significantly increased in the NPC model group transfected with CK13 gene and the treatment group injected with CK13 gene. When the mice were killed on the 9th day, the tumor inhibition rates of the experimental group were 44.6 and 47.1and 45.9%, respectively. The inhibition rate of the empty carrier group was 0 ~ 0.05. The difference between each experimental group and the empty carrier group was significant (p 0.05). The difference was not statistically significant (P 0.05). The positive expression rate of CK13 in the experimental group was higher than that in the empty vector group and the control group. The expression level of CK13mRNA in the experimental group was higher than that in the empty vector and the control group. Western blotting showed that there were obvious CK13 protein bands in the experimental group, but there were no obvious bands in the empty vector group and the control group. Conclusion the NPC gene may be a new target gene for tumor therapy and provide a new approach for gene therapy of nasopharyngeal carcinoma.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.63

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