基于SIRT6介导有氧糖酵解-ROS-自噬信号轴探讨枳实多甲氧基黄酮逆转结肠癌耐药的机制研究

结肠癌因化疗易耐药而预后差，如何逆转耐药是亟待解决的临床问题，中医认为耐药和脾虚关系密切。枳实是健脾常用中药，中医典籍已有其抗肿瘤的功效记载，且已得到现代药理学研究的证实，但尚无PMFs逆转结肠癌耐药的机制研究。受SIRT6调控的有氧糖酵解-ROS-自噬信号轴在耐药中发挥重要作用。我们发现PMFs能抑制HCT8/T细胞的有氧糖酵解、减少ROS过度产生并抑制自噬小体的形成，从而逆转结肠癌耐药；敲低HCT8/T的SIRT6蛋白水平，P-gp及Nrf2的表达降低，细胞凋亡率增加，而PMFs能抑制SIRT6蛋白表达。因此，我们推测：PMFs是通过SIRT6介导的有氧糖酵解-ROS-自噬信号轴逆转结肠癌耐药。我们拟从分子、细胞、动物水平探究：（1）PMFs逆转结肠癌耐药的体内外作用机制；（2）PMFs逆转耐药的体内药效及药动学特征。为“有瘤体必虚，有虚首健脾”提供实验依据，进而为临床应用提供新思路。

Colon cancer is easily resistant to chemotherapy and has a poor prognosis, so reversing drug resistance is urgently needed. Chinese medicine believes that colon cancer drug resistance is closely related with spleen function. Aurantii fructus immaturus is a traditional Chinese medicine commonly used to strengthen the spleen, there are many records of antitumor effect about aurantii fructus immaturus extract in medical books, and it has been confirmed in modern pharmacological studies. However, there are few studies on reversal of colon cancer resistance by PMFs, and the study on mechanism of action of PMFs is even less.Aerobic glycolysis-ROS- autophagy signal axis played an important role in tumor drug resistance, and this signaling axis may be regulated by SIRT6. PMFs was found to be a good reversal agent for colon cancer drug resistance in early experiments. PMFs can inhibit the aerobic glycolysis of HCT8/T cells, reduce the excessive production of ROS and inhibit the formation of autophagosomes. When the SIRT6 protein level of HCT8/T cells was knocked down, the protein levels of P-gp and Nrf2 were also significantly decreased, and the apoptosis rate of HCT8/T cells was increased, while PMFs could inhibit the SIRT6 protein expression of HCT8/T cells. Therefore, we hypothesized that PMFs reverses colon cancer drug resistance through SIRT6-mediated aerobic glycolysis-ROS-autophagy signaling axis. This project aims to study from the molecular, cellular and animal levels in order to clarify that:(1) The in vivo and in vitro mechanism of PMFs reversing drug resistance of colon cancer; (2) The in vivo efficacy and pharmacokinetics of PMFS in reversing drug resistance of colon cancer. It provides experimental basis for“Tumor can cause debilitating, the first choice is to strengthen the spleen”, and further provides new ideas and theoretical reference for clinical application.