目的:研究fat—1基因在人乳腺癌细胞内的表达、功能及其对乳腺癌细胞增殖的影响。方法:把fat-1基因插入到腺病毒的穿梭载体中,与骨架载体同源重组,构建腺病毒重组载体(Ad.GFP.fatl),将通过包装细胞系(293)产生的腺病毒感染人乳腺癌株QMB2细胞。提取细胞的总RNA,以fat-1的反义mRNA作探针,用NorthernBlot检测fat-1基因在人乳腺癌株QMR2细胞内的表达。流式细胞仪分析n-3脂肪酸脱氢酶对人乳腺癌株QMB2细胞增殖的影响。气象色谱仪分析n-3脂肪酸脱氢酶对人乳腺癌株QMR2细胞的n-6PUFAs/n-3PUFAs含量影响。结果:fat-1基因在人乳腺癌株QMR2细胞中能有效异源表达,2d后检测到fat-1mRNA的条带。fat-1基因抑制了人乳腺癌株QMB2细胞的增殖,降低了20%(P<0.05);同时降低了人乳腺癌株QMR2细胞n-6PUFAs/n-3PUFAs含量降低。结论:腺病毒介导的fat-1基因能在人乳腺癌株QMR2细胞内有效异源表达,且抑制人乳腺癌株QMR2细胞的增殖。

Objective: To transfer the gene of n-3 fatty acid desaturase fat-1 into human breast cancer cell QMR2 by adenovirus vector and study the effect of the gene on proliferation of QMR2 cells. Methods: The gene fat-1 was cloned into the shuttle vector of adenovirus, and homologously recombined with an adenoviral backbone vector ( pAdEasy 1) to generate the recombinant adenovirus Ad. GFP. fall; the virus was packaged in 293 cells, inoculated on the breast cancer cells QMR2; total RNA of the cells was hybridized with antisense RNA of fat-1 mRNA by Northern to analyze the expression of fat-1; the effect of fat-1 on the proliferation of QMR2 cells was analyzed by Flow Cytometry; the content of n-6 PUFAs/n-3 PUFAs was analyzed by Gas Chromatography. Results: The high titer recombinant virus was got through DNA recombinant ; the fat-1 mRNA appeared in breast cancer cell QMR2 after virus Ad. GFP. fat1 infected the cells for 2 days; compared with the control cells (Ad. GFP) , proliferation of QMR2 cells was inhibited by the gene fat-1, decreased by 31% ( P < 0.05); moreover, fat-1 gene decreased content of n-6 PUFAs/n-3 PUFAs. Conclusion: The gene fat - 1 was heter-ologously expressed in human breast cancer cell QMR2 via adenovirus, and the expression produced an inhibitory effect on proliferation of the cells.

R730.5

国家自然科学基金(NO．30271217) 青岛市科技局科技计划项目(No．03-1-YN-14)