改变细胞膜的脂肪酸组成可促进乳腺癌细胞凋亡

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改变细胞膜的脂肪酸组成可促进乳腺癌细胞凋亡

侯琳耿芳宋薛美兰王秀丽刘颖葛银林

青岛大学医学院生物化学与分子生物学教研室青岛大学医学院生物化学与分子生物学教研室青岛大学医学院生物化学与分子生物学教研室青岛大学医学院生物化学与分子生物学教研室青岛大学医学院生物化学与分子生物学教研室青岛大学医学院生物化学与分子生物学教研室

Changing Composition of Fatty Acids of Cell Membrane Promotes

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摘要： 目的: 研究n-6脂肪酸脱氢酶 fat-1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fat-1 基因的重组腺病毒载体 (Ad.GFP.fat-1)，通过包装细胞系（293）产生的腺病毒，感染人乳腺癌细胞MCF-7。提取细胞的总RNA，以fat-1的反义mRNA 作探针，用Northern Blot检测fat-1 基因在MCF-7细胞内的表达。MTT法分析fat-1 基因对MCF-7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MCF-7细胞的n-6 PUFAs/n-3 PUFAs含量影响。结果: 通过基因重组技术，得到预期的重组病毒；fat-1 基因在人乳腺癌细胞MCF-7 中能有效异源表达，2天后，可检测到fat-1 mRNA的条带。与对照细胞相比，fat-1基因有效地抑制了MCF-7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MCF-7细胞膜n-6 PUFAs/n-3 PUFAs的比率。结论: 腺病毒介导的fat-1 基因能在人乳腺癌细胞MCF-7内有效异源表达，且抑制了MCF-7细胞的增殖。机理为降低了细胞膜的n-6 PUFAs/n-3 PUFAs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.

收稿日期: 2005-09-22 出版日期: 2006-05-25

通讯作者: 葛银林

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引用本文:

侯琳,耿芳宋,薛美兰,王秀丽,刘颖,葛银林. 改变细胞膜的脂肪酸组成可促进乳腺癌细胞凋亡[J]. 中国生物工程杂志, .

. Changing Composition of Fatty Acids of Cell Membrane Promotes. China Biotechnology, .

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https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2006/V26/I05/38

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