Effect of eicosapentaenoic acid on the proliferation and incidence of apoptosis in the colorectal cell line HT29
Effect of eicosapentaenoic acid on the proliferation and incidence of apoptosis in the colorectal cell line HT29
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DOI：
10.1007/s11745-999-0480-7
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发表时间：
1999-12-01
期刊：
影响因子：
1.9
通讯作者：
Johnson, IT
中科院分区：
文献类型：
--
作者：
Clarke, RG;Lund, EK;Johnson, IT
鱼油已被证明可以通过抑制有丝分裂、增加细胞凋亡或两者兼而有之的机制来减少动物模型中结直肠癌的诱发。我们使用人结肠腺癌细胞系HT29来探讨长链n-3多不饱和脂肪酸二十碳五烯酸（ESA）对体外细胞增殖和死亡的影响。细胞在含有 5、10 和 15 μg/mL EPA 的培养基中培养。使用细胞数量和胸苷掺入来量化增殖，并使用流式细胞术研究细胞周期效应。使用细胞电泳、膜联蛋白-V 结合和形态学标准来表征细胞凋亡。贴壁细胞和自由漂浮的分离细胞被视为两个不同的群体。在 10 和 15 μg/mL EPA 存在下，由于贴壁细胞的脱离增加，贴壁 HT29 集落的生长速率显着降低。用 10 或 15 μg/mL EPA 处理后，S 期贴壁细胞的比例增加，表明 S 期晚期或 G2 早期阻断。漂浮细胞显示出广泛的DNA裂解的证据，但在用10或15μg/mL EPA处理时，尽管漂浮细胞的数量增加，但具有亚CO DNA含量的漂浮细胞的比例下降。我们得出的结论是，PA 不会抑制贴壁细胞的有丝分裂，但会增加它们与基质分离的速度，可能是在细胞凋亡开始的早期阶段。这种机制可能类似于“失巢凋亡”，或因细胞接触丧失而诱导细胞凋亡，并且可能有助于鱼油在体内的抗癌作用。
Fish oil has been shown to reduce the induction of colorectal cancer in animal models by a mechanism which may involve suppression of mitosis, increased apoptosis, or both. We used the human colonic adenocarcinoma cell line HT29 to explore the effects of the long-chain n-3 polyunsaturated fatty acid eicosapentaenoic acid (ESA) on cell proliferation and death in vitro. Cells were cultured in media containing EPA at 5, 10, and 15 mu g/mL. Cell number and thymidine incorporation were used to quantify proliferation, and cell cycle effects were studied using flow cytometry. Cel electrophoresis, annexin-V binding, and morphological criteria were used to characterize apoptosis. Adherent cells and freely floating detached cells were treated as two distinct populations. In the presence of EPA at 10 and 15 mu g/mL there was a marked reduction in the growth rate of adherent HT29 colonies, owing to an increased detachment of adherent cells. After treatment with 10 or 15 mu g/mL EPA the proportion of adherent cells in S-phase increased, indicating either a block in late S-phase or early G2. Floating cells showed evidence of extensive DNA cleavage, but the proportion of floating cells with sub CO DNA content declined on treatment with 10 or 15 mu g/mL EPA even though the number of floating cells increased. We conclude that PA does not inhibit mitosis of adherent cells, but increases the rate at which they become detached from the substrate, probably at an early stage in the initiation of apoptosis. This mechanism may be analogous to "anoikis," or induction of apoptosis in response to loss of cell contact, and may contribute to the anticarcinogenic effects of fish oil in vivo.