| 张明明 徐玉清△ 尚娜娜 崔凤 姜礼红 孟佳 李文静.NK细胞中过表达Livin的实验研究[J].,2016,16(30):5824-5828 |
| NK细胞中过表达Livin的实验研究 |
| Overexpression of Livin Gene in Natural Killer Cells |
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| DOI： |
| 中文关键词: 自然杀伤细胞 质粒 Livin 转染 |
| 英文关键词: Natural killer cells Vector Livin Transfection |
| 基金项目:吴阶平医学基金会基金资助（320.6751.1215） |
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| 中文摘要: |
| 目的：将携带Livin 的质粒pIRES2-EGFP-Livin 进行扩增，转染自然杀伤细胞(NK)及胃癌细胞株SGC-7901，并检测其在NK
及胃癌细胞株SGC-7901 中的表达。方法：将携带Livin 基因的质粒pIRES2-EGFP-Livin 进行扩增，鉴定质粒纯度与浓度；从健康
人外周血中获得NK细胞，应用HP 转染试剂将质粒pIRES2-EGFP-Livin 转染体外培养的NK及胃癌细胞，对比分析NK 及胃癌
细胞株SGC-7901 中基因转染效率及目的基因的表达情况。结果：用无血清培养基在体外成功的扩增大量的NK细胞；质粒提取
试剂盒抽提得到大量无内毒素的质粒，质粒DNA 基因序列并未发生突变，浓度和纯度较高。胃癌细胞株SGC-7901 中观察到明显
的质粒pIRES2-EGFP-Livin 绿色荧光表达；而NK中未观察到绿色荧光表达。结论：质粒pIRES2-EGFP-Livin 能使Lvin 蛋白表达
于胃癌细胞株SGC-7901 中，而在NK 中未表达。 |
| 英文摘要: |
| Objective:To amplify the plasmid pIRES2-EGFP-Livin encoding functional human Livin gene to transfect natural
killer cell (NK) and gastric carcinoma cell line SGC-7901, and test the expression in NK and gastric carcinoma cell line SGC-7901.Methods:The plasmid pIRES2-EGFP-Livin encoding functional human livin gene was amplified, the concentration and the purity of
plasmid pIRES2-EGFP-Livin were identified. NK from peripheral blood of healthy people were acquired And the plasmid pIRES2-
EGFP-Livin was transfected into NK and gastric carcinoma cell line SGC-7901 using HP transfection reagent in vitro. The transfection
efficiency and the expression of human livin in NK and gastric carcinoma cell line SGC-7901 were compared.Results:A large quantity
of NK cells were successfully amplified by serum-free medium in vitro; amounts of no endotoxin plasmid were acquired from Plasmid
Midi Kit; plasmid had no mutations with high concentration and purity. The green bands of EGFP expression could be clearly observed in
pIRES2-EGFP-Livin-transfected gastric carcinoma cell line SGC-7901, while could not be observed in NK.Conclusion:Effective
expressions of Livin protein had been verified in gastric carcinoma cell line SGC-7901 transfected by Plasmid pIRES2-EGFP-Livin but
not in NK. |
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