Department of Pharmacy, Fujian Provincial Maternal and Child Health Hospital, Fuzhou, Fujian 350001, China.
Department of Pharmacy, Fujian Provincial Maternal and Child Health Hospital, Fuzhou, Fujian 350001, China.
Life Sci. 2014 Mar 18;99(1-2):44-51. doi: 10.1016/j.lfs.2014.01.066. Epub 2014 Feb 7.
Cardamonin has previously demonstrated that it had an antiproliferative effect on vascular smooth muscle cells by inhibiting the activity of mammalian target of rapamycin (mTOR). The antiproliferative effect and the possible mechanism of combining with mTOR of cardamonin were investigated on A549 cells.
Cell proliferation, cell cycle and apoptosis were measured by methyl thiazolyl tetrazolium (MTT) and flow cytometry, respectively. mTOR and 12 kDa FK506 binding protein (FKBP12) were transfected into A549 cells by Lipofectamine. Western blots were used to examine the mTOR expressions and its activities, and the expressions of 70 kDa ribosomal S6 kinase (p70S6K), FKBP12 and Interleukin-2 (IL-2), respectively.
Treated with cardamonin, the proliferation of A549 cells was inhibited. Meanwhile, cell cycle was blocked and DNA synthesis was decreased whereas cell apoptosis was promoted, and the activation of mTOR and p70S6K was decreased by cardamonin. Transfected with mTOR or FKBP12, proliferation of A549 cells was increased. Rapamycin had a similar degree of effect on antiproliferation of both transfected cells. However, the antiproliferative effect of cardamonin on mTOR transfected cells was stronger than that on FKBP12 transfected cells. Both rapamycin and cardamonin decreased the phosphorylation of mTOR and p70S6K in two kinds of transfected cells. Cardamonin had no effect on the expression of FKBP12 and IL-2, whereas the expressions were decreased by rapamycin.
Cardamonin inhibited proliferation and induced apoptosis of A549 cells via mTOR. It might directly interact with mTOR independently of binding with FKBP12.
小豆蔻明通过抑制哺乳动物雷帕霉素靶蛋白（mTOR）的活性，先前已证明其对血管平滑肌细胞具有抗增殖作用。研究了小豆蔻明对 A549 细胞的 mTOR 结合的抗增殖作用及其可能的机制。
通过噻唑蓝（MTT）和流式细胞术分别测定细胞增殖、细胞周期和细胞凋亡。用脂质体将 mTOR 和 12 kDa FK506 结合蛋白（FKBP12）转染入 A549 细胞。用 Western blot 检测 mTOR 的表达及其活性，以及 70 kDa 核糖体 S6 激酶（p70S6K）、FKBP12 和白细胞介素-2（IL-2）的表达。
小豆蔻明处理可抑制 A549 细胞的增殖。同时，细胞周期阻滞，DNA 合成减少，细胞凋亡增加，小豆蔻明可降低 mTOR 和 p70S6K 的激活。转染 mTOR 或 FKBP12 后，A549 细胞的增殖增加。雷帕霉素对转染细胞的增殖抑制具有相似的作用。然而，小豆蔻明对 mTOR 转染细胞的增殖抑制作用强于 FKBP12 转染细胞。雷帕霉素和小豆蔻明均可降低两种转染细胞中 mTOR 和 p70S6K 的磷酸化。小豆蔻明对 FKBP12 和 IL-2 的表达无影响，而雷帕霉素可降低其表达。
小豆蔻明通过 mTOR 抑制 A549 细胞增殖并诱导其凋亡。它可能直接与 mTOR 相互作用，而不与 FKBP12 结合。
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