Does melatonin induce apoptosis in MCF-7 human breast cancer cells in vitro?
Does melatonin induce apoptosis in MCF-7 human breast cancer cells in vitro?
复制标题
DOI：
10.1034/j.1600-079x.2002.1821.x
复制
发表时间：
2002-03-01
影响因子：
10.3
通讯作者：
Sánchez-Barceló, EJ
中科院分区：
文献类型：
--
作者：
Cos, S;Mediavilla, MD;Sánchez-Barceló, EJ
褪黑激素抑制雌激素反应性 MCF-7 人乳腺癌细胞的增殖。这项工作的目的是评估褪黑激素是否不仅可以调节 MCF-7 细胞增殖，还可以诱导细胞凋亡。在本实验中，我们使用 1,25-二羟基胆钙化醇 (D-3) 作为阳性对照，因为它抑制 MCF-7 细胞增殖并诱导细胞凋亡。 MCF-7 细胞与 I rim 褪黑激素、100 nM D-3 或其稀释液一起培养，以确定它们对细胞增殖、细胞活力、细胞周期阶段分布、凋亡细胞群以及 p53、p21WAF1、bcl-2、bcl-X-L 和 bax 蛋白表达的影响。孵育 24 或 48 小时后，与对照组相比，褪黑素和 D-3 处理均显着减少了活细胞数量，但在处理之间未观察到细胞活力的差异。细胞凋亡的发生率（以落入 DNA 直方图亚 G(1) 区域的细胞群或通过 TUNEL 反应测量）在褪黑激素处理的细胞和对照细胞中相似，而正如预期的那样，用 D-3 处理的细胞中的细胞凋亡高于对照细胞。与褪黑激素或 D-3 孵育 24 或 48 小时后，p53 和 p21WAF1 蛋白的表达显着增加。用褪黑素处理后，bcl-2、bcl-X-L 和 bax mRNA 没有检测到显着变化，而在 D-3 处理的细胞中，观察到 bcl-X-L 显着下降。这些数据支持这样的假设：褪黑激素通过控制 p53-p21 途径调节细胞周期长度来减少 MCF-7 细胞增殖，但不会明显诱导细胞凋亡。
Melatonin inhibits proliferation of the estrogen-responsive MCF-7 human breast cancer cells. The objective of this work was to assess whether melatonin not only regulates MCF-7 cell proliferation but also induces apoptosis. In this experiment we used 1,25-dihydroxycholecalciferol (D-3) as a positive control because it inhibits MCF-7 cell proliferation and induces apoptosis. MCF-7 cells were cultured with either I rim melatonin, 100 nM D-3 or its diluent to determine their effects on cell proliferation, cell viability, cell-cycle phase distribution, population of apoptotic cells, and expression of p53, p21WAF1, bcl-2, bcl-X-L and bax proteins. After 24 or 48 hr of incubation, both melatonin and D-3-treatment significantly decreased the number of viable cells in relation to the controls, although no differences in cell viability were observed between the treatments. The incidence of apoptosis, measured as the population of cells falling in the sub-G(1) region of the DNA histogram, or by the TUNEL reaction, was similar in melatonin-treated and control cells whereas, as expected, apoptosis was higher among cells treated with D-3 than in controls. The expression of p53 and p21WAF1 proteins significantly increased after 24 or 48 hr of incubation with either melatonin or D-3. No significant changes in bcl-2, bcl-X-L and bax mRNAs were detected after treatment with melatonin whereas in D-3-treated cells, a significant drop in bcl-X-L was observed. These data support the hypothesis that melatonin reduces MCF-7 cell proliferation by modulating cell-cycle length through the control of the p53-p21 pathway, but without clearly inducing apoptosis.