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

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。

Construction of Fat-1 Eukaryotic Expression Vector and Its Effect on Fatty Acids Content of Human Oral Squamous Cell Carcinoma Cells

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.