节点文献
细胞膜脂肪酸组成改变促进乳腺癌细胞凋亡的研究
Changing the Composition of Fatty Acids of Cell Membrane Promotes Breast Cancer Cell to Apoptosis
【摘要】 目的:研究n6脂肪酸脱氢酶fat1基因在人乳腺癌细胞内的表达,改变细胞膜脂肪酸组成,对乳腺癌细胞的凋亡作用。方法:构建含有fat1基因的重组腺病毒载体(Ad.GFP.fat1),通过包装细胞系(293)产生的腺病毒,感染人乳腺癌细胞MCF7。提取细胞的总RNA,以fat1的反义mRNA作探针,用Northernblot检测fat1基因在MCF7细胞内的表达。MTT法分析fat1基因对MCF7细胞增殖的影响,凋亡染色试剂盒检测细胞的凋亡。气相色谱仪分析对MCF7细胞的n6PUFAs/n3PUFAs含量影响。结果:通过基因重组技术,得到预期的重组病毒;fat1基因在人乳腺癌细胞MCF7中能有效异源表达,2d后,可检测到fat1mRNA的条带。与对照细胞相比,fat1基因有效地抑制了MCF7细胞的增殖(23%,p<0.05),促进了凋亡(增加35%);同时降低了人乳腺癌细胞MCF7细胞膜n6PUFAs/n3PUFAs的比率。结论:腺病毒介导的fat1基因能在人乳腺癌细胞MCF7内有效异源表达,且抑制了MCF7细胞的增殖。机理为降低了细胞膜的n6PUFAs/n3PUFAs的比率。
【Abstract】 Objective: To study expression of the gene of n-6 fatty acid desaturase fat-1 in human breast cancer cell, composition change of fatty acids of cell membrane, and effect of the gene on apoptosis of breast cancer cell. Methods: Construct recombinant adenovirus vector (Ad.GFP.fat-1) containing fat-1 gene, the recombinant adenovirus was produced in 293 package cell, then it infected the breast cancer cells MCF-7; Total RNA of the cells was isolated and hybridized with antisense RNA probe of fat-1 mRNA by Northern to analyze the expression of fat-1 in MCF-7;The effect of fat-1on the proliferation of MCF-7 cell was analyzed by MTT method,apoptosis of the cells were detected by apoptosis kit;Content of n-6 PUFAs/n-3 PUFAs was analyzed by Gas Chromatography. Results: The proposed recombinant virus was got through DNA recombinant technique; fat-1 gene effectively expressed in human breast cancer cell MCF-7; The fat-1 mRNA band appeared 2 days after infection of virus Ad.GFP.fat-1;Compared with the control cell (Ad.GFP), proliferation of MCF-7 cell was markedly inhibited by the gene fat-1( 23%, p<0.05) and apoptosis of the cells was promoted;Moreover, fat-1 gene decreased ratio of n-6 PUFAs/n-3 PUFAs of MCF-7 cell membrane. Conclusion: The fat-1gene transfer mediated by adenovirus could heterologously express in human breast cancer cell MCF-7, inhibit proliferation of the cell. The mechanism analysis shown that fat-1 decreased ration of n-6 PUFAs/n-3 PUFAs of cell membrane.
- 【文献出处】 中国生物工程杂志 ,China Biotechnology , 编辑部邮箱 ,2006年05期
- 【分类号】R737.9
- 【被引频次】2
- 【下载频次】238