蔓荆子黄素通过HIF-1α调控糖代谢抑制非小细胞肺癌细胞的增殖  

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作　　者：危靖怡 宁辉 董佳琪 韩乐 陈文娟 雷光焰 WEI Jing-yi;NING Hui;DONG Jia-qi;HAN Le;CHEN Wen-juan;LEI Guang-yan(Shaanxi University of Chinese Medicine,Xianyang 710046,China;Department of Thoracic Surgery,Shaanxi Provincial Cancer Hospital,Xi'an 710061,China;Department of Medical Oncology,Shaanxi Provincial Cancer Hospital,Xi'an 710061,China)

机构地区：[1]陕西中医药大学,陕西咸阳710046 [2]陕西省肿瘤医院胸部肿瘤外科,陕西咸阳710061 [3]陕西省肿瘤医院肿瘤内科,陕西咸阳710061

出　　处：《中国中药杂志》2024年第24期6755-6762,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81902321);陕西省重点研发计划一般项目(2022SF-261)。

摘　　要：研究蔓荆子黄素对非小细胞肺癌细胞H322增殖的影响,并对其分子机制进行探究。首先采用cell counting kit-8(CCK-8)实验、克隆形成和EdU实验检测浓度和作用时间梯度的蔓荆子黄素对H322细胞增殖能力的作用。接着检测蔓荆子黄素作用前后H322细胞的葡萄糖摄取、乳酸生成、细胞外pH和细胞耗氧量,分析蔓荆子黄素对非小细胞肺癌细胞H322糖酵解的影响。最后进一步采用实时荧光定量PCR(RT-qPCR)和Western blot实验探究蔓荆子黄素作用的糖酵解相关分子机制。实验表明,蔓荆子黄素抑制非小细胞肺癌细胞H322的增殖,且与药物剂量和作用时间呈正相关,H322细胞给药48、72 h后,其半数抑制浓度(IC_(50))分别为28.64、19.41μmol·L^(-1)。同时蔓荆子黄素抑制H322细胞的葡萄糖摄取和乳酸生成,并使细胞外pH和细胞耗氧量增加。进一步探究发现蔓荆子黄素抑制糖酵解相关分子葡萄糖转运蛋白1(GLUT1)、己糖激酶2(HK2)、醛缩酶A(ALDOA)、丙酮酸激酶M2型(PKM2)以及缺氧诱导因子-1α(HIF-1α)的表达。最后验证了过表达HIF-1α可以逆转蔓荆子黄素对H322细胞增殖和糖酵解的抑制。以上结果提示,蔓荆子黄素可能通过抑制HIF-1α的表达来调控细胞糖酵解,从而抑制非小细胞肺癌H322的增殖。该研究为非小细胞肺癌的治疗发掘了具有潜力的药物,并提供了研究方向。The study investigated the effect of casticin on the proliferation of non-small cell lung cancer(NSCLC) H322 cells and explored its molecular mechanism. Firstly, the cell counting kit-8(CCK-8) assay, colony formation assay, and EdU assay were used to detect the effect of casticin on the proliferation capacity of H322 cells under different concentrations and treatment durations. Then, glucose uptake, lactate production, extracellular pH, and oxygen consumption of H322 cells were measured before and after casticin treatment to analyze its impact on glycolysis in NSCLC H322 cells. Finally, real-time fluorescence quantitative PCR(RT-qPCR) and Western blot assays were performed to explore glycolysis-related molecules affected by casticin. The experiments showed that casticin inhibited the proliferation of NSCLC H322 cells in a dose-and time-dependent manner, with half-maximal inhibitory concentrations(IC_(50)) of 28.64 and 19.41 μmol·L^(-1) after 48 and 72 hours of treatment, respectively. Casticin also inhibited glucose uptake and lactate production in H322 cells, while increasing extracellular pH and oxygen consumption. Further investigation revealed that casticin inhibited the expression of glycolysis-related molecules, including glucose transporter 1(GLUT1), hexokinase 2(HK2), aldolase A(ALDOA), pyruvate kinase M2(PKM2), and hypoxia-inducible factor-1α(HIF-1α). Overexpression of HIF-1α was found to reverse the inhibitory effects of casticin on H322 cell proliferation and glycolysis. These findings suggest that casticin may regulate cellular glycolysis by inhibiting the expression of HIF-1α, thereby inhibiting the proliferation of NSCLC H322 cells. This study identifies a potential drug for the treatment of NSCLC and provides a direction for further research.

关 键 词：蔓荆子黄素 非小细胞肺癌 HIF-1Α 增殖 糖代谢重编程 

分 类 号：R28[医药卫生—中药学]