| 李栋聂代邦王海军段新平逄大欣△ 欧阳红生.Fat-1 基因真核表达载体的构建及其对人口腔鳞癌细胞
脂肪酸含量的影响[J].,2011,11(11):2117-2121 |
| Fat-1 基因真核表达载体的构建及其对人口腔鳞癌细胞
脂肪酸含量的影响 |
| Construction of Fat-1 Eukaryotic Expression Vector and Its Effect on FattyAcids Content of Human Oral Squamous Cell Carcinoma Cells |
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| DOI： |
| 中文关键词: 真核表达载体 转染 口腔鳞癌细胞 n-3 脂肪酸 |
| 英文关键词: Eukaryotic expression vector Transfect Oral squamous cell carcinoma cell n -3 fatty acids |
| 基金项目:国家级转基因生物新品种培育重大专项（2008ZX08006-003） |
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| 中文摘要: |
| 目的：构建真核表达载体pcDNA3.1-Fat1，线性化稳定转染人口腔鳞癌细胞株Tca8113，检测其细胞内脂肪酸含量变化。方
法：通过重叠延伸PCR 方法人工合成利于真核表达的Fat-1 基因，用基因重组技术构建真核表达载体pcDNA3.1-Fat-1，用脂质体
转染真核细胞的方法转染人口腔鳞癌细胞株Tca8113，用气相色谱仪检测脂肪酸的变化情况。结果：测序及酶切鉴定成功合成真
核偏好表达的Fat-1 基因。与对照组相比，转染Fat-1 基因的口腔癌细胞的n-3 脂肪酸明显增多，n-6/n-3 明显下降。结论：成功构建
真核表达载体pcDNA3.1-Fat1，并对口腔鳞癌细胞内脂肪酸含量产生明显影响，为进一步研究Fat-1 基因在口腔鳞癌中的生物学
功能奠定了基础。 |
| 英文摘要: |
| Objective: To construct eukaryotic expression vector pcDNA3.1-Fat1 and transfect human oral squamous cell
carcinoma cell line Tca8113, then tested the changes of fatty acids content in Tca8113. Methods: The Fat-1 gene was constructed by
SOE-PCR. The pcDNA3.1-Fat1 was constructed by using recombinant DNA technology, human oral squamous carcinoma cells line
Tca8113cells was transfected using lipofection method. The fatty acids content of the transfected Tca8113 cells was detected by using gas
chromatography technology. Results: Eukaryotic expression vector pcDNA3.1-Fat1 was successfully constructed and transfected into
human oral squamous carcinoma cell lineTca8113. The cells that expressed the fat-1 gene had a lower n-6/n-3 PUFA ratio compared with
the cells that expressed the control vector. Conclusion: The pcDNA3.1-Fat1 is successfully constructed, and it has significant effect on the
PUFAs content of Tca8113 cells, which makes a foundation in the future study on the biological function of Fat-1 gene in human oral
squamous carcinoma. |
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