| 王辉区裕升沈晗黄树林△.TCR BV12-3 重组载体构建及其抗肿瘤作用的初步研究*[J].,2014,14(2):206-208 |
| TCR BV12-3 重组载体构建及其抗肿瘤作用的初步研究* |
| Construction and Primary Study on the Anti-Tumor Effect of TCR BV12-3Recombinant Vector* |
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| DOI： |
| 中文关键词: T 细胞受体 BV12-3 克隆 肿瘤 |
| 英文关键词: TCR BV12-3 Clone Tumor |
| 基金项目:国家“重大新药创制”科技重大专项(2009ZX09103-708)；广东省医学科研课题（B2002070） |
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| 中文摘要: |
| 摘要目的：构建pEGFP-C3-TCR BV12-3 表达载体并初步研究其抗肿瘤作用。方法：TCR BV12-3 基因片段从pGEM-T-TCR
BV12-3 载体上酶切并克隆至pEGFP-C3 载体中，通过脂质体将pEGFP-C3-TCR BV12-3 表达载体转染外周血单个核细胞
（PBMCs），48 小时后荧光显微镜观察转染效率。PBMCs细胞、pEGFP-C3 载体转染的PBMCs 细胞、pEGFP-C3-TCR BV12-3 表达
载体转染的PBMCs 细胞分别与肝癌细胞BEL-7402 和宫颈癌细胞HeLa 共培养24 h，显微镜观察肿瘤细胞的生长情况。结果：测
序证实TCR BV12-3 基因片段成功亚克隆至pEGFP-C3 载体中，荧光显微镜证实重组体转染PBMCs 细胞48 h后可有效表达绿
色荧光。显微镜观察发现pEGFP-C3-TCR BV12-3载体转染的PBMCs 对肝癌细胞有杀伤作用，但对宫颈癌杀伤作用不明显。结
论：成功构建pEGFP-C3-TCR BV12-3 表达载体，初步证实TCRBV12-3 对肝癌细胞有杀伤作用。 |
| 英文摘要: |
| ABSTRACT Objective：To construct pEGFP-C3-TCR BV12-3 vector, and to investigate the effect of TCR BV12-3 on anti-tumor.
Methods：TCR BV12-3 gene was digested from pGEM-T-TCR BV12-3, and was cloned into pEGFP-C3 vector. The pEGFP-C3-TCR
BV12-3 recombinant vector was transfected into peripheral blood mononuclear cells (PBMCs) via liposome. After 48 h, the transfection
efficiency was measured by fluorescence microscope. Then, the PBMCs, pEGFP-C3-TCR BV12-3 and pEGFP-C3 transfection were
co-cultured with hepatocarcinoma cells BEL-7402 and cervical cancer cells HeLa, respectively. After 24 h incubation, the anti-tumor
effect of TCR BV12-3 was investigated by microscope. Results：The DNA sequencing analysis showed that TCR BV12-3 gene was
successfully subcloned into pEGFP-C3 vector, and the green fluorescence could be detected by fluorescence microscope in PBMCs
transfected with pEGFP-C3-TCR BV12-3. PBMCs transfected with pEGFP-C3-TCR BV12-3 showed cytotoxicity in BEL-7402 cells.
Conclusion：The pEGFP-C3-TCR BV 12-3 vector was successfully constructed, and the anti-tumor effect of TCR BV12-3 was proved
initially. |
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