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  本文選題：CK13基因 + 鼻咽癌��； 參考：《昆明醫(yī)科大學(xué)》2012年碩士論文

【摘要】：目的：本文觀察CK13基因?qū)θ吮茄拾﹦游锬Ｐ头派涿舾行缘挠绊�。本課題是在前期研究基礎(chǔ)上,利用HNE1細(xì)胞株和轉(zhuǎn)染了目的基因的HNEl細(xì)胞株在Nu-Nu裸鼠腋下分別建立荷瘤動物模型,即NPC動物模型組和含目的基因的動物模型組,并在NPC動物模型組瘤體部位直接注射多價陽離子脂質(zhì)體包裹表達(dá)CK13基因的質(zhì)粒,對照組分別注射空載體質(zhì)粒和磷酸鹽緩沖液,然后所有組均給予2Gy直線加速器的局部腫瘤放射治療(簡稱放療),4天后重復(fù)腫瘤局部注射和放療。觀察腫瘤治療前后大小變化,計(jì)算抑瘤率。最后處死小鼠,分離腫瘤組織。采用免疫組織學(xué)染色和RT-PCR的方法檢測腫瘤組織CK13表達(dá)情況,HE染色比較各組移植瘤組織壞死程度。Western blotting進(jìn)行CK13蛋白定量分析。探討CK13基因與鼻咽癌放射敏感性的關(guān)系,以CK13基因?yàn)榘悬c(diǎn)進(jìn)行相關(guān)研究,可為臨床提高鼻咽癌放療敏感性提供新的思路。 方法：1.NPC動物模型及含目的基因NPC動物模型的建立。2.NPC動物模型局部注射質(zhì)粒。3.放療,并測量放療前后腫瘤大小。4.處死小鼠,取標(biāo)本做病檢、免疫組化、RT-PCR、及Western Blotting。5.結(jié)果進(jìn)行統(tǒng)計(jì)學(xué)分析。 結(jié)果：在轉(zhuǎn)染CK13基因的NPC動物模型組和注射有CK13基因的治療組中CK13表達(dá)水平明顯升高,CK13基因聯(lián)合放療治療組腫瘤生長明顯受限制。在第9天處死小鼠時,實(shí)驗(yàn)組(A1、A2、B1、B2組)的抑瘤率分別為44.6%,47.1%,45.9%,49.5%、空載體組(C1、C2組)抑瘤率為0,0.05%,各個實(shí)驗(yàn)組分別與空載體組比較,差異均有統(tǒng)計(jì)學(xué)差異(p0.05),各個實(shí)驗(yàn)組間相比較,差異無統(tǒng)計(jì)學(xué)意義(p0.05)。免疫組化檢查實(shí)驗(yàn)組CK13陽性表達(dá)率較空載體組和對照組高,且差異有統(tǒng)計(jì)學(xué)意義。RT-PCR檢測CK13基因顯示：實(shí)驗(yàn)組CK13mRNA表達(dá)水平較空載體和對照組高。Western blotting示：實(shí)驗(yàn)組均可見明顯CK13蛋白條帶,而空載體組和對照組未見明顯條帶。 結(jié)論：CK13基因可提高NPC對放射治療的敏感性；CK13基因可能成為腫瘤治療一個新的靶基因,為鼻咽癌的基因治療提供一個新的途徑。
[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.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R739.63

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