靶向激活腺苷酸活化蛋白激酶（AMPK）诱导结肠癌相关多个关键促癌基因降解的实验机制研究

Our preliminary studies have shown that multiple oncogenes including EGFR, PDGFRα/β and Fascin1 (FSCN1) are over-expressed and/or over-activated in colorectal cancers, and they are important for cancer progression and metastasis. AMPK activation induces lysosomal degradation of above oncogenes in colon cancer cells. Meanwhile, we indentified two novel AMPK activators including the anti-fungi drug itraconazole and antagomiR-451. Both of them activated AMPK signaling while inhibiting cancer cell growth. Thus, we propose that AMPK activation-mediated anti-colon cancer efficiency may be associated with lysosomal degradation of above key oncogenes. In the current proposal, by using multiple bio-medical assays, we will study the underlying mechanism of above oncogenes' degradation by AMPK activation. We are set to test the anti-colon cancer ability of the two newly indentified AMPK activators, itraconazole and antagomiR-451, both in vivo and in vitro. We will also test the expressions p-ACC (an indicator of AMPK activation) and above oncogenes in human colon cancer tissues, and analyze their relationship with cancer prognosis. This study suggests that AMPK activation-induced anti-colon cancer efficiency might be associated with its ability to induce degradation of multiple key oncogenes. Meanwhile, the two AMPK activators, itraconazole and antagomiR-451, might be further tested in clinical settings for anti-colon cancer therapy.

我们前期研究发现活化AMPK的抗结肠癌作用可能与其诱导多个关键促癌基因（EGFR、PDGFRα/β和Fascin-1）降解有关，提示活化AMPK可能是一种有效提高抗肿瘤药物疗效的新策略。目前已知蛋白降解主要有溶酶体依赖和蛋白酶体依赖途径，但结肠癌中激活AMPK降解上述促癌基因的具体机制尚未明确。我们已有研究表明溶酶体酶及细胞自噬抑制剂可逆转AICAR(AMPK激活剂)诱导的上述促癌基因降解，但结肠癌中关键促癌基因高表达是否与AMPK通路及溶酶体降解有关还需进一步明确。据此，我们提出本课题的假说：结肠癌细胞中，靶向激活AMPK通过活化自噬－溶酶体降解途径，导致上述相关多个关键促癌基因降解。为验证该假设，本课题拟通过体内、外实验探讨活化AMPK诱导上述关键促癌基因溶酶体转位的分子机制，为新型AMPK活化剂伊曲康唑和antagomiR-451发挥抗结肠癌作用提供理论和实验依据。

靶向激活腺苷活化蛋白激酶（AMPK）具有抗结肠癌的作用，活化AMPK的抗结肠癌作用可能与其诱导多个关键促癌基因（EGFR、PDGFRα/β和Fascin-1）降解有关，提示活化AMPK可能是一种有效提高抗肿瘤药物疗效的新策略。研究表明溶酶体酶及细胞自噬抑制剂可逆转AICAR(AMPK激活剂)诱导的上述促癌基因降解，但结肠癌中关键促癌基因高表达是否与AMPK通路及溶酶体降解有关还需进一步明确。本课题拟通过体内、外实验探讨活化AMPK诱导上述关键促癌基因溶酶体转位的分子机制，并通过C6神经酰胺的增加通过活化AMPK激活p53的通路，增敏长春新碱的抗结肠癌作用。此外，我们发现多个AMPK活化剂：C6神经酰胺、冬凌草甲素、Itraconazole、icaritin、Aqueous Oldenlandia diffusa extracts、mTORC1/2抑制剂INK-128抗癌的分子机制，利用分子、细胞生物学等方法，体内外靶向调节腺苷酸活化蛋白激酶（AMPK）抗结肠癌作用，明确诱导结肠癌相关多个关键促癌基因（oncogene）降解的机制，阐明AMPK信号通路在其中的作用，并调节癌细胞内神经酰胺含量和AMPK活化水平，达到增敏抗癌作用的目的，为开发增敏抗肿瘤类药物提供理论和实验依据。