青蒿琥酯通过HAND2激活LKB1-AMPK-mTOR信号通路抑制雌激素介导的子宫内膜癌的机制研究

The incidence of endometrial cancer is rising in the world. The applicant's research suggests that artesunate plays a certain role in the prevention and treatment of endometrial cancer, but the mechanism is not yet clear. In the preliminary experiment, we found artesunate can increase HAND2 expression in endometrial carcinoma cell line, and our previous studies have found that HAND2 gene is low expressed in endometrial carcinoma, and HAND2 and LKB1-AMPK-mTOR signaling pathway are closely related to estrogen receptor (ER-a). This study speculates that artesunate may play roles through ER-a by HAND2, and inhibition of ER-a may inhibit the occurrence of endometrial cancer through activation of LKB1-AMPK-mTOR signaling pathway. Based on the previous work, we prepare to further investigate the effects of artesunate in estrogen mediated endometrial cancer by activation of LKB1-AMPK-mTOR signaling pathway by HAND2 from multi-level systems including animal, cellular and molecular level, thus revealing the molecular mechanisms and the targets of artesunate in endometrial carcinoma.

子宫内膜癌的发病率在世界范围内逐年上升。申请者的前期研究提示青蒿琥酯在子宫内膜癌的防治中起一定作用，但作用机制尚未明确。预实验中，申请者发现青蒿琥酯能使子宫内膜癌细胞株中的心脏神经脊衍生物表达转录因子2（HAND2）表达量升高，而申请者前期发现HAND2基因在子宫内膜癌中低表达，且HAND2和LKB1-AMPK-mTOR信号通路均与雌激素受体（ER-a）密切相关。本研究推测青蒿琥酯可能通过HAND2介导ER-α发挥作用，并且通过抑制ER-α可能激活LKB1-AMPK-mTOR通路抑制子宫内膜癌的发生。我们拟在前期工作的基础上，从动物、细胞和分子水平多层次系统深入探讨青蒿琥酯通过HAND2激活LKB1-AMPK-mTOR信号通路在雌激素介导的子宫内膜癌中的作用，揭示青蒿琥酯抗子宫内膜癌的分子机制和作用靶点。

本研究通过体外细胞实验证实青蒿琥酯对雌激素介导的子宫内膜癌细胞具有抑制增殖、促进细胞凋亡的作用，但对迁移的抑制作用不明显，并具有浓度依赖性。裸鼠移植瘤实验也证实青蒿琥酯可抑制子宫内膜癌的生长。通过建立过表达HAND2和干扰HAND2的稳转株，应用ER-α抑制剂、LKB1抑制剂，我们发现青蒿琥酯在ER-α阳性的子宫内膜癌中可能是通过HAND2-LKB1-AMPK-mTOR信号轴来发挥抑制子宫内膜癌的作用，进一步的HAND2 基因敲除小鼠模型结果也显示HAND2可激活LKB1-AMPK-mTOR 信号通路在雌激素介导的子宫内膜癌的发挥作用。此外，通过检测子宫内膜癌及正常子宫内膜组织标本中HAND2、LKB1、AMPK、mTOR的表达，我们进一步证实了这些分子的相关性。根据以上结果，我们得出结论：青蒿琥酯可通过HAND2激活LKB1-AMPK-mTOR信号通路在雌激素介导的子宫内膜癌中发挥抑制作用。