一种新的咪唑通过AMPK/mTOR途径诱导肿瘤细胞自噬

A New Kind of Imidazoles Induces Tumor Cell Autophagy via AMPK/mTOR Pathway

自噬与肿瘤的发生有着密切的关系.通过防止受损蛋白质和细胞器的毒性积累,自噬限制氧化应激、慢性组织损伤和致癌信号传导,抑制癌症的发生,这表明了自噬激活在癌症预防和治疗中的作用.研究发现一种新的咪唑,命名为NO.143.利用胰腺癌细胞PANC-1为模型,用MTS检测不同浓度NO.143对细胞活力的影响,共聚焦显微镜观察自噬荧光点,Western blot检测自噬相关蛋白LC3B、ATG13、p62的表达情况,以及AMPK/mTOR信号通路相关蛋白的表达情况.结果表明,NO.143呈现浓度依赖性地抑制肿瘤细胞的增殖.NO.143能够明显增加细胞中自噬荧光斑点、LC3B表达、ATG13的含量和p62的降解,表明NO.143能够显著上调肿瘤细胞的自噬.进一步研究显示,NO.143能够增加AMPKα的磷酸化水平,降低哺乳动物雷帕霉素靶蛋白(mTOR)和S6的磷酸化水平.相反的,在抑制AMPKα活性后,这种现象发生逆转.这些结果都可能表明NO.143通过AMPK/mTOR信号传导途径诱导肿瘤自噬,抑制肿瘤细胞的增殖.

Autophagy is closely related to the tumor occurrence.By preventing the accumulation of toxicities in damaged proteins and organelles,autophagy limits oxidative stress,chronic tissue damage and oncogenic signaling,inhibiting the development of cancer.This indicates the role of autophagy activation in cancer prevention and treatment.It was found a new imidazole named NO.143.PANC-1 was used as a model to detect the effect of different concentrations of NO.143 on cell viability by MTS.The autophagy fluorescence spots were observed by confocal microscopy.Western blot was used to detect the expression of autophagy-related proteins LC3B,ATG13,p62 and the expression of AMPK/mTOR signaling pathway-related proteins.The results showed that NO.143 showed a concentration-dependent inhibition of tumor cell proliferation.NO.143 can significantly increase autophagic fluorescent spots in cells,increase LC3B expression,ATG13 content and p62 degradation.These results show that NO.143 can significantly up-regulate autophagy in tumor cells.Further studies showed that NO.143 was able to increase the phosphorylation level of AMPKαand reduce the phosphorylation levels of mammalian rapamycin target protein(mTOR)and S6.Conversely,this phenomenon reverses after inhibiting AMPKαactivity.These results may indicate that NO.143 induces tumor autophagy through the AMPK/mTOR signaling pathway and inhibits tumor cell proliferation.