人Man2cl基因表达对移植性肿瘤在小鼠的生长及小鼠体液性免疫应答的影响

发布时间：2018-01-17 20:19

本文关键词：人Man2cl基因表达对移植性肿瘤在小鼠的生长及小鼠体液性免疫应答的影响　出处：《中国协和医科大学》2006年硕士论文　论文类型：学位论文

【摘要】： MAN2C1是本实验室克隆的cDNA编码的1个人α-甘露糖苷酶。我们原先称它为6A8α-甘露糖苷酶。国际已命名此酶为mannosidase，alpha，class 2C，member 1[Homo sapiens]，简称MAN2C1。其相应的基因位于染色体15q11-q13。HUGO基因命名委员会命名此基因为Man2c1。MAN2C1的分子量在非糖基化情况下为116kDa。为了在整体水平研究MAN2C1的生物学功能，本实验室用显微注射线性pIRES2-EGFP-MAN2C1的方法，制备了转Man2c1基因的ICR小鼠。本论文工作用基因组PCR的方法检测Man2c1基因整合入小鼠基因组的情况。在113只原代小鼠中，共检测到8只阳性小鼠。经配对繁殖得到F1代小鼠。基因组PCR检测见～1／2小鼠为阳性。阳性F1代小鼠合笼在F2代有～3／4小鼠阳性。F3代小鼠约有80％-90％的阳性。用RT-PCR检测了Man2c1基因在转基因鼠尾部组织中的转录，在7个系的小鼠中有4个为Man2c1 mRNA阳性。这表明，已成功制备了转Man2c1基因小鼠。在35#转基因小鼠观测到毛稀疏的表型变化。本论文工作研究了Man2c1基因表达对移植性肿瘤生长及对体液性免疫的影响。在小鼠荷瘤实验中，观察到转基因小鼠的肿瘤生长明显快于非转基因小鼠，且肿瘤在前者呈浸润性生长，并有引流淋巴结转移，，而在后者，大部分肿瘤有完整的包膜，也无淋巴结转移。这表明转入的Man2c1基因表达促进肿瘤生长。在对相关机制的探索中，观察到转基因小鼠脾脏单个核细胞中的NK细胞百分率增高，但脾脏单个核细胞对YAC-1细胞的杀伤无变化。在体液性免疫实验中，小鼠用BSA免疫，见转基因小鼠的脾脏大于非转基因小鼠，但血清抗BSA抗体的滴度却低于非转基因鼠。为进一步了解MAN2C1的生物学意义，本论文工作还藉RNAi技术抑制小鼠NIH-3T3细胞的Man2c1基因表达，探索了Man2c1基因表达对细胞在培养中生长的影响。但未见阳性结果。
[Abstract]:MAN2C1 is a human 伪 -mannosidase encoded by cDNA in our laboratory. We originally called it 6A8 伪 -mannosidase. It has been named mannosidase internationally. Alpha,class 2C,member 1. [Homo sapiens]. The corresponding gene is located on chromosome 15q11-q13.HUGO gene nomenclature committee named this gene because the molecular weight of Man2c1.MAN2C1 is in the non-glycosylation condition. Under the condition of 116kDa. In order to study the biological function of MAN2C1 at the whole level, the microinjection of linear pIRES2-EGFP-MAN2C1 was used in our laboratory. ICR mice with Man2c1 gene were prepared. In this paper, the integration of Man2c1 gene into mouse genome was detected by genomic PCR. A total of 8 positive mice were detected. F1 generation mice were obtained by paired breeding. Genomic PCR analysis showed that 1 / 2 / 2 mice were positive. About 80% to 90% of the mice were positive. RT-PCR was used to detect the transcription of Man2c1 gene in the tail tissue of transgenic mice. Four of the 7 strains were Man2c1 mRNA positive. Transgenic mice with Man2c1 gene have been successfully prepared. The phenotypic changes of hair sparsity were observed in 3 transgenic mice. In this paper, the effect of Man2c1 gene expression on the growth of transplanted tumors and on humoral properties was studied. Immune effects. In mice tumor-bearing experiments. It was observed that the tumor growth of transgenic mice was much faster than that of non-transgenic mice, and the tumor had infiltrative growth and lymph node metastasis in the former, while in the latter, most of the tumors had complete capsule. There was also no lymph node metastasis, which indicated that the Man2c1 gene expression promoted tumor growth. In the exploration of the related mechanism, the percentage of NK cells in splenic mononuclear cells of transgenic mice was increased. In humoral immunity test, mice were immunized with BSA, and the spleen of transgenic mice was larger than that of non-transgenic mice. But the titer of serum anti-BSA antibody was lower than that of non-transgenic mice. In order to further understand the biological significance of MAN2C1, the expression of Man2c1 gene in mouse NIH-3T3 cells was inhibited by RNAi technique. The effect of Man2c1 gene expression on cell growth was explored, but no positive results were found.
【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】：2006
【分类号】：R392;R73-3

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