氟伐他汀通过P53/AMPK/mTOR通路诱导自噬抑制人肺腺癌细胞骨转移的实验研究

肺癌骨转移是肺癌患者最严重的并发症之一，当前的治疗方式尚不能获得满意效果。近年发现他汀类药物能抑制多种肿瘤生长及转移，但其作用机的分子机制尚未明确。我们的前期研究提示氟伐他汀能抑制肺腺癌细胞迁移和侵袭，在裸鼠动物模型体内能抑制肺腺癌骨转移，并提示与诱导自噬有关。本课题通过Western blot及免疫荧光检测自噬标志蛋白LC3-II/LC3-I 、Beclin-1的表达及自噬体的形成，并使用3-methyladenine、Bafilomycin抑制自噬反应，观察裸鼠肺腺癌骨转移变化，论证氟伐他汀是否通过诱导自噬抑制肺腺癌骨转移。其次，以p53/AMPK/mTOR通路为主要研究目标，观察氟伐他汀对mTOR活性的影响，在使用Pifithrin-α及siRNA干扰P53表达后，检测AMPK及mTOR活性变化及与自噬反应的关系，探讨氟伐他汀诱导自噬的主要信号通路，以期探寻肺腺癌骨转移的防治新策略。

Bone metastasis is one of the most serious complications of lung cancer patients, and current treatment strategies are not satisfying yet. Recent year, it has been reported that fluvastatin can inhibit the growth and metastasis of a variety of tumor types, but the molecular mechanism is unclear so far. Our previous findings showed that fluvastatin inhibited lung cancer cells migration and invasion and prevented bone metastases in lung cancer in nude mice, the mechanism of which could be related to autophagy. In this project, we intend to examine whether fluvastatin can trigger autophagy by detecting autophagy markers LC3-II / LC3-I and Beclin-1 expression with Western blot analysis and autophagosome formation with fluorescence tracing, and if so, to investigate in nude mice whether fluvastatin-induced bone metastases inhibition results from autophagy by using autophagy inhibitor, 3-methyladenine and bafilomycin. Furthermore, we plan to focus on p53/AMPK/mTOR pathway to validate the effect of fluvastatin on mTOR activity and role of p53 in regulating AMPK, mTOR and autophagy activity by p53 inhibitor pifithrin-α and p53 siRNA. In this research, we expect to reveal the key signaling pathways involved in fluvastatin-triggered autophagy and find a new strategy for prevention and treatment of bone metastases in lung cancer.