兰州大学机构库 >学院待认领
小鼠肾癌Delta样分子4疫苗的初步研究
Alternative TitleA pilot study on Delta-like gland 4 as a DNA vaccine in mice Renal Cell Carcinoma
何綦琪
Thesis Advisor岳中瑾
2012-05-23
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
KeywordDLL4 裸鼠 肾癌 疫苗
Abstract目的: 初步研究制备DLL4 疫苗并验证在裸鼠肾癌模型上的作用机制。制备表达DLL4的质粒,并观察在DLL4免疫后的裸鼠上种植肿瘤后的生长情况、生长大小及局部变化等,为肾癌生物免疫疗法及临床辅助肾癌治疗提供一定的实验依据。 方法: 1.DLL4免疫载体的制备: DLL4 cDNA是由特异性引物PCR扩增得到DLL4开放阅读表达框,限制性内切酶双酶切后插入同样酶切的pVAX1质粒并经酶切和测序验证,纯化后转染细胞进行Western免疫印记验证。 2.裸鼠免疫: 4周龄的雄性裸鼠随机分为2组,皮下注射超纯化的pVAX1-DLL4载体悬液;另一组为其阴性对照,注射空白pVAX1载体悬液,分3周次免疫裸鼠,40ug/次。 3.裸鼠肾癌模型的建立及免疫效果评价: 以皮下注射形式接种肾癌细胞于2组裸鼠腋下及腹股沟旁(用量为:0.2ml/只),拟观察肿瘤的生长情况及其4周及8周后肿瘤直径,做随机对照分组实验结果分析其免疫效果。 结果: DLL4 cDNA表达框序列为2058 bp,转入pVAX1质粒载体后经酶切和测序验证构建成功; 对照组部分裸鼠在种植肿瘤细胞系后3周生出肿瘤,肿瘤较实验组大,约(1.26±0.91)cm,由此得出可以使用裸鼠建立肾癌动物模型; 实验组DLL4肿瘤疫苗免疫的小鼠肿瘤生长较小,大小为(1.07±0.70)cm,肿瘤外观略发红。 结论: 1.可以通过PVAX1连接DLL4制备成含有DLL4的质粒作为肿瘤疫苗。 2.使用裸鼠可以成功建立动物肾细胞癌模型。
Other AbstractObjective: To develop the rational DLL4 cDNA vaccine to renal cell carcinoma and the mechanism of the vaccine on nude mice RCC models, we have thus developed the study to explore the possible immunization by DLL4-encoding plasmid DNA vaccine to evaluate the vaccine efficacy by some nude mice admitted to the RCC. Method: The development of DLL4-encoding plasmid: Using some specific primers to get the DLL4 express box by PCR, we had a restrictive analysis, inserted pVAX1 plasmids and did sequencing verification, purificated the transfected cells and had a Western bolt to carry out the DLL4 cDNA clone. Results: The DLL4 cDNA expression box has a length of 2058bp. Sequence verification and restricted enzyme cutting sites were proved after it was transfected into PVAX1 plasmids.
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/222493
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
何綦琪. 小鼠肾癌Delta样分子4疫苗的初步研究[D]. 兰州. 兰州大学,2012.
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