一种新型中药配方通过触发磷酸戊糖途径依赖的氧化应激抑制非小细胞肺癌

A Novel Chinese Medicine Formula Inhibits Non-small Cell Lung Cancer by Triggering Oxidative Stress Dependent on Pentose Phosphate Pathway

背景与目的非小细胞肺癌(non-small cell lung cancer,NSCLC)是世界范围内死亡率较高的恶性肿瘤之一。一种新型中药配方(novel Chinese medicine formula-01,NCHF-01)在治疗NSCLC方面已显示出显著的临床疗效,但此配方治疗NSCLC的作用机制尚不完全清楚。本研究拟探讨中药配方抑制NSCLC的作用及其分子机制。方法建立Lewis肺癌细胞(lewis lung cells,LLC)荷瘤小鼠,检测NCHF-01抑制肿瘤的效果;苏木素-伊红(hematoxylin-eosin staining,HE)染色检测LLC荷瘤小鼠组织器官的形态变化;NCHF-01处理NSCLC细胞,MTT和结晶紫染色实验检测其对细胞活力和增殖的影响;流式细胞术检测细胞周期、凋亡和活性氧(reactive oxygen species,ROS)水平;网络药理学预测其抑制NSCLC的作用机制;Western blot和免疫组织化学(immunohistochemistry,IHC)检测相关蛋白的表达。结果NCHF-01能够抑制LLC荷瘤小鼠肿瘤的生长,并对其他组织器官无明显毒副作用;NCHF-01能够抑制细胞活力和增殖,诱导细胞的G2/M期停滞和细胞凋亡,促进细胞内ROS水平的升高;网络药理学分析显示,NCHF-01通过氧化应激和中枢碳代谢等多种生物学过程发挥抗NSCLC作用;NCHF-01能够降低磷酸戊糖途径(pentose phosphate pathway,PPP)关键酶6-磷酸葡萄糖脱氢酶(6-phosphate glucose dehydrogenase,G6PD)和6-磷酸葡萄糖酸脱氢酶(6-phosphogluconate dehydrogenase,6PGD)的蛋白表达以及酶活性。结论NCHF-01能够通过PPP依赖的氧化应激抑制NSCLC。

Background and objective Non-small cell lung cancer(NSCLC)is one of the most lethal malignan-cies worldwide.A novel Chinese medicine formula-01(NCHF-01)has shown significant clinical efficacy in the treatment of NSCLC,but the mechanism of this formula in the treatment of NSCLC is not fully understood.The aim of this study is to in-vestigate the molecular mechanism of NCHF-01 in inhibiting NSCLC.Methods Lewis lung cells(LLC)tumor bearing mice were established to detect the tumor inhibitory effect of NCHF-01.The morphological changes of tissues and organs in LLC tumor-bearing mice were detected by hematoxylin-eosin(HE)staining.NSCLC cells were treated by NCHF-01.The effects of cell viability and proliferation were detected by MTT and crystal violet staining experiment.Flow cytometry was used to detect cell cycle,apoptosis and reactive oxygen species(ROS).Network pharmacology was used to predict the mechanism of its inhibitory effect of NSCLC.Western blot and immunohistochemistry(IHC)were used to detect the expression of related proteins.Results NCHF-01 can inhibit tumor growth in LLC tumor-bearing mice,and has no obvious side effects on other tissues and organs.NCHF-01 could inhibit cell viability and proliferation,induce G2/M phase arrest and apoptosis,and promote the increase of ROS level.Network pharmacological analysis showed that NCHF-01 exerts anti-NSCLC effects through various biological processes such as oxidative stress and central carbon metabolism.NCHF-01 can reduce the protein expres-sion and enzyme activity of the key enzymes 6-phosphate glucose dehydrogenase(G6PD)and 6-phosphogluconate dehydro-genase(6PGD)in the pentose phosphate pathway(PPP).Conclusion NCHF-01 can inhibit NSCLC through oxidative stress dependent on the PPP.