Fatty acid modulation of epidermal growth factor-induced mouse mammary epithelial cell proliferation in vitro.
Fatty acid modulation of epidermal growth factor-induced mouse mammary epithelial cell proliferation in vitro.
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表皮生长因子诱导的小鼠乳腺上皮细胞体外增殖的脂肪酸调节。
DOI：
10.1006/excr.1994.1243
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发表时间：
1994
影响因子：
3.7
通讯作者：
Briski,KP
中科院分区：
文献类型：
--
作者：
Sylvester,PW;Birkenfeld,HP;Hosick,HL;Briski,KP
从怀孕中期的BALB/c小鼠中分离乳腺上皮细胞，在胶原凝胶中进行原代培养，并在含10 ng/ml表皮生长因子（EGF）的无血清培养基中维持。与未处理的对照组相比，添加饱和脂肪酸na -硬脂酸钠（18:0）的培养基显著减弱了乳腺上皮细胞的增殖，而添加不饱和脂肪酸na -花生四烯酸钠（20:4）的培养基显著增强了乳腺上皮细胞的增殖。用不同剂量的18:0或20:4处理也发现导致乳腺上皮细胞膜脂肪酸组成的直接剂量依赖性富集，并同时通过气相色谱测定其他膜脂肪酸的相对水平降低。前列腺素合成抑制剂吲哚美辛在所有治疗组中均显著抑制egf诱导的细胞生长，但没有改变脂肪酸治疗的相对抑制（18:0）或刺激（20:4）作用。与对照组相比，egf诱导的PKC向乳腺上皮细胞膜部分的易位在20:4时增强，在18:0时减弱。磷脂依赖性蛋白激酶C同工酶的Western blot分析表明，原代培养的小鼠乳腺上皮细胞中主要存在PKCα同工酶，该同工酶的分子量为85 kDa。这些结果表明，添加特定脂肪酸的培养基与乳腺上皮细胞膜脂肪酸组成、PKC激活和有丝分裂反应性的显著改变有关。由于EGF可以诱导PKC激活和细胞增殖，并且PKC激活需要膜源磷脂和二酰基甘油，因此这些数据表明，脂肪酸对乳腺上皮细胞有丝分裂发生的特异性调节是通过PKCα激活的改变来介导的。
Mammary epithelial cells were isolated from mid-pregnant BALB/c mice, grown in primary culture within collagen gels, and maintained with serum-free medium containing 10 ng/ml epidermal growth factor (EGF) as the mitogen. Supplementation of culture medium with the saturated fatty acid, Na-stearate (18:0), significantly attenuated, whereas treatment with the unsaturated fatty acid, Na-arachidonate (20:4), significantly enhanced mammary epithelial cell proliferation, as compared to untreated controls. Treatment with various doses of either 18:0 or 20:4 was also found to result in a direct dose-dependent enrichment of mammary epithelial cell membrane fatty acid composition and a concurrent decrease in the relative levels of other membrane fatty acids, as determined by gas chromatography. Administration of the prostaglandin synthesis inhibitor, indomethacin, significantly inhibited EGF-induced cell growth in all treatment groups, but did not alter the relative inhibitory (18:0) or stimulatory (20:4) effects of fatty acid treatment. EGF-induced PKC translocation into the membrane fraction of mammary epithelial cells was enhanced in 20:4 and attenuated in 18:0 treatment groups, as compared to controls. Western blot analysis of phospholipid-dependent protein kinase C isoenzymes showed that PKCαwas the predominant isoenzyme present in mouse mammary epithelial cells grown in primary culture, and the molecular weight of this PKC isoenzyme was determined to be 85 kDa. These results suggest that supplementation of culture media with specific fatty acids is associated with significant alterations in mammary epithelial cell membrane fatty acid composition, PKC activation, and mitogenic responsiveness. Since EGF can induce both PKC activation and cell proliferation, and because PKC activation requires membrane-derived phospholipids and diacylglycerol, these data suggest that specific fatty acid modulation of mammary epithelial cell mitogenesis is mediated through alterations in PKCαactivation.