College of Korean Medicine, Kyung Hee University, Seoul, Republic of Korea.
Institute of Sports Science, Kyung Hee University, Yongin, Republic of Korea.
Phytother Res. 2023 Sep;37(9):4224-4235. doi: 10.1002/ptr.7903. Epub 2023 May 26.
To elucidate the underlying antitumor mechanism of lambertianic acid (LA) derived from Pinus koraiensis, the role of cancer metabolism related molecules was investigated in the apoptotic effect of LA in DU145 and PC3 prostate cancer cells. MTT assay for cytotoxicity, RNA interference, cell cycle analysis for sub G1 population, nuclear and cytoplasmic extraction, lactate, Glucose and ATP assay by ELISA, Measurement of reactive oxygen species (ROS) generation, Western blotting, and immunoprecipitation assay were conducted in DU145 and PC3 prostate cancer cells. Herein LA exerted cytotoxicity, increased sub G1 population and attenuated the expression of pro-Caspase3 and pro-poly (ADP-ribose) polymerase (pro-PARP) in DU145 and PC3 cells. Also, LA reduced the expression of lactate dehydrogenase A (LDHA), glycolytic enzymes such as hexokinase 2 and pyruvate kinase M2 (PKM2) with reduced production of lactate in DU145 and PC3 cells. Notably, LA decreased phosphorylation of PKM2 on Tyr105 and inhibited the expression of p-STAT3, cyclin D1, C-Myc, β-catenin, and p-GSK3β with the decrease of nuclear translocation of p-PKM2. Furthermore, LA disturbed the binding of p-PKM2 and β-catenin in DU145 cells, which was supported by Spearman coefficient (0.0463) of cBioportal database. Furthermore, LA generated ROS in DU145 and PC3 cells, while ROS scavenger NAC (N-acetyl L-cysteine) blocked the ability of LA to reduce p-PKM2, PKM2, β-catenin, LDHA, and pro-caspase3 in DU145 cells. Taken together, these findings provide evidence that LA induces apoptosis via ROS generation and inhibition of PKM2/β-catenin signaling in prostate cancer cells.
为了阐明源自红松的 Lambertianic 酸（LA）的抗肿瘤机制，研究了癌症代谢相关分子在 LA 诱导 DU145 和 PC3 前列腺癌细胞凋亡中的作用。采用 MTT 法检测细胞毒性，RNA 干扰，亚 G1 期细胞周期分析，核质提取，ELISA 法检测乳酸、葡萄糖和 ATP 水平，检测活性氧（ROS）生成，Western blot 法和免疫沉淀法检测 DU145 和 PC3 前列腺癌细胞。结果显示，LA 发挥细胞毒性作用，增加 DU145 和 PC3 细胞的亚 G1 期细胞群，并减弱 pro-Caspase3 和 pro-poly（ADP-ribose）polymerase（pro-PARP）的表达。此外，LA 降低乳酸脱氢酶 A（LDHA）和糖酵解酶（如己糖激酶 2 和丙酮酸激酶 M2（PKM2））的表达，减少乳酸在 DU145 和 PC3 细胞中的产生。值得注意的是，LA 降低 Tyr105 上 PKM2 的磷酸化，并抑制 p-STAT3、细胞周期蛋白 D1、C-Myc、β-catenin 和 p-GSK3β的表达，同时减少核内 p-PKM2 的转位。此外，LA 扰乱了 DU145 细胞中 p-PKM2 和 β-catenin 的结合，这一结果得到了 cBioportal 数据库 Spearman 系数（0.0463）的支持。此外，LA 在 DU145 和 PC3 细胞中产生 ROS，而 ROS 清除剂 NAC（N-乙酰-L-半胱氨酸）阻断了 LA 降低 p-PKM2、PKM2、β-catenin、LDHA 和 pro-caspase3 的能力在 DU145 细胞中。综上所述，这些发现提供了证据，表明 LA 通过 ROS 生成和抑制前列腺癌细胞中 PKM2/β-catenin 信号通路诱导细胞凋亡。
