Javrushyan Hayarpi, Ginovyan Mikayel, Harutyunyan Tigran, Gevorgyan Smbat, Karabekian Zaruhi, Maloyan Alina, Avtandilyan Nikolay
Research Institute of Biology, Yerevan State University, Yerevan, Armenia.
Department of Genetics and Cytology, Yerevan State University, Armenia.
PLoS One. 2025 Apr 3;20(4):e0303736. doi: 10.1371/journal.pone.0303736. eCollection 2025.
Plants of the Hypericaceae family have been traditionally used for their medicinal properties, including antibacterial, antiviral, and antioxidant activities. Among these, Hypericum alpestre (HA) extracts have shown notable cytotoxicity against various cancer cell lines, drawing attention to their phenolic compounds as potential anticancer agents. Similarly, N(G)-Nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase (NOS) activity, has emerged as a promising candidate in cancer therapy. However, the precise molecular mechanisms underlying the anticancer effects of both HA and L-NAME remain unclear. This study aimed to clarify the impact of HA and L-NAME on the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/ mammalian target of rapamycin (mTOR) signaling pathway in A549 human lung adenocarcinoma and MDA-MB-231 breast cancer triple-negative cells, with particular emphasis on the tumor necrosis factor-alpha (TNFα)/cyclooxygenase-2 (COX-2) and vascular endothelial growth factor A (VEGFα)/matrix metalloproteinase-2 (MMP-2) pathways. In silico analyses identified compounds within HA extracts with the highest affinity for PI3K/Akt, a finding subsequently confirmed by in vitro experiments. Notably, the combination of HA and L-NAME demonstrated greater efficacy than the combination of HA and 5-fluorouracil (5-FU), as evidenced by enhanced apoptotic activity. Both HA alone and in combination with L-NAME inhibited the TNFα/COX-2 and VEGFα/MMP-2 pathways. These results suggest that the therapeutic effects of HA, especially in combination with L-NAME, may be mediated through the PI3K/Akt signaling pathway. A better understanding of the interaction between HA polyphenols and PI3K/Akt signaling could pave the way for novel therapeutic strategies against cancer, including drug-resistant tumors.
金丝桃科植物传统上因其药用特性而被使用，包括抗菌、抗病毒和抗氧化活性。其中，阿尔卑斯金丝桃（HA）提取物对多种癌细胞系显示出显著的细胞毒性，其酚类化合物作为潜在的抗癌剂受到关注。同样，一氧化氮合酶（NOS）活性抑制剂N（G）-硝基-L-精氨酸甲酯（L-NAME）已成为癌症治疗中有前景的候选药物。然而，HA和L-NAME抗癌作用的确切分子机制仍不清楚。本研究旨在阐明HA和L-NAME对A549人肺腺癌和MDA-MB-231乳腺癌三阴性细胞中磷酸肌醇3-激酶（PI3K）/蛋白激酶B（Akt）/雷帕霉素哺乳动物靶蛋白（mTOR）信号通路的影响，特别关注肿瘤坏死因子-α（TNFα）/环氧化酶-2（COX-2）和血管内皮生长因子A（VEGFα）/基质金属蛋白酶-2（MMP-2）通路。计算机分析确定了HA提取物中对PI3K/Akt具有最高亲和力的化合物，这一发现随后得到体外实验的证实。值得注意的是，HA和L-NAME的组合比HA和5-氟尿嘧啶（5-FU）的组合显示出更高的疗效，凋亡活性增强证明了这一点。单独使用HA以及与L-NAME联合使用均抑制了TNFα/COX-2和VEGFα/MMP-2通路。这些结果表明，HA的治疗作用，尤其是与L-NAME联合使用时，可能通过PI3K/Akt信号通路介导。更好地理解HA多酚与PI3K/Akt信号之间的相互作用可为包括耐药肿瘤在内的新型抗癌治疗策略铺平道路。
