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目的观察N-6/N-3多不饱和脂肪酸(POLYUNSATURATED FATTY ACID,PUFA)不同比例对乳腺癌细胞缝隙连接细胞通讯的影响。方法N-6/N-3PUFA不同比例(1~10)处理MCF-7(ER+)和MDA-MB-231(ER-)乳腺癌细胞,采用免疫细胞化学观察细胞核KI-67表达,WESTERN BLOT检测细胞膜和细胞质缝隙连接蛋白CX26、CX43表达,划痕负载染料迁移法分析细胞缝隙连接通讯功能。结果免疫组化结果显示,单纯N-6和10∶1N-6/N-3PUFA增强两种乳腺癌细胞KI-67表达(P<0.05),5∶1N-6/N-3PUFA虽无明显变化,但单纯N-3和1∶1N-6/N-3PUFA抑制细胞KI-67表达(P<0.01)。免疫印迹发现,单纯N-3和1∶1N-6/N-3PUFA能明显上调MCF-7细胞膜和细胞质的CX26、CX43蛋白表达,单纯N-6PUFA却下调两种蛋白表达;在MDA-MB-231细胞,N-6/N-3PUFA不同比例对膜和细胞质的CX43蛋白表达与MCF-7细胞具有相同趋势,但对CX26蛋白则无明显作用。用划痕负载染料迁移法却发现,N-6/N-3PUFA不同比例对细胞缝隙连接通讯功能均无明显影响。结论单纯N-3和1∶1N-6/N-3PUFA对MCF-7和MDA-MB-231乳腺癌细胞增殖活性的抑制作用,可能是CX26和CX43蛋白通过独立于缝隙连接细胞通讯功能的调节所致。
Objective To investigate the effect of different ratios of N-6 / N-3 polyunsaturated fatty acids (PUFA) on the gap junctional cell junctions in breast cancer cells. Methods KI-67 expression in nuclei of MCF-7 (ER +) and MDA-MB-231 (ER-) breast cancer cells was detected by immunocytochemistry in different proportions of N-6 / Cell membrane and cytoplasmic connexin CX26, CX43 expression, scratch-loaded dye migration assay of cell gap junctional communication. Results Immunohistochemistry showed that N-6 and 10:1 N-6 / N-3 PUFAs enhanced KI-67 expression in both breast cancer cell lines (P <0.05) , But N-3 alone and 1: 1N-6 / N-3PUFA inhibited cell KI-67 expression (P <0.01). Western blotting showed that N-3 and 1: 1N-6 / N-3PUFA could significantly up-regulate the expression of CX26 and CX43 in the membrane and cytoplasm of MCF-7 cells, while N-6PUFA alone down-regulated the expression of both proteins. 231 cells and N-6 / N-3 PUFAs, the expression of CX43 protein in membrane and cytoplasm had the same tendency as that of MCF-7 cells, but had no significant effect on CX26 protein. Scratch-loading dye migration method found that different ratios of N-6 / N-3PUFA had no significant effect on cell gap junctional communication. Conclusion The inhibitory effect of N-3 alone and 1: 1 N-6 / N-3 PUFA on the proliferation of MCF-7 and MDA-MB-231 breast cancer cells may be due to the regulation of the CX26 and CX43 proteins independently of the gap junction cell communication Due.