Sayilan Ozgun Gulben, Ozgun Eray, Karabas Tugce, Suer Gokmen Selma, Eskiocak Sevgi
Department of Medical Biochemistry, Trakya University School of Medicine, Edirne, 22030, Turkey.
Naunyn Schmiedebergs Arch Pharmacol. 2024 Dec 21. doi: 10.1007/s00210-024-03725-0.
We aimed to determine the effects of piperine on cell viability, cellular stresses, and apoptosis first, then the relationship of piperine's effects with the c-Jun N-terminal kinase (JNK) signaling pathway, and also the interaction of piperine with sorafenib in hepatocellular carcinoma. Hepatocellular carcinoma (HepG2 and Hep3B) and non-cancerous hepatocyte (AML12) cell lines were used. The cell viability was determined by using MTT assay. Cellular stresses, apoptosis, and JNK signaling markers were measured by Western blotting. Cells were pre-treated with SP600125 as a JNK inhibitor. The inhibitory concentration 50% (IC50) values and interaction of piperine with sorafenib were calculated by using CompuSyn software. IC50 values of piperine were 97 µM for HepG2, 58 µM for Hep3B, and 184 µM for AML12 with incubation for 48 h. Piperine caused a significant concentration-dependent increase in cellular stresses, apoptosis, and activated JNK signaling in hepatocellular carcinoma cells. Pre-treatment with a JNK inhibitor significantly reduced piperine-induced cellular stresses, apoptosis, and cytotoxicity. Piperine had concentration-dependent additive or synergistic effects when combined with sorafenib in both HepG2 and Hep3B cells. We found that piperine induces cellular stresses, apoptosis, and cytotoxicity via JNK signaling and has concentration-dependently additive or synergistic effects with sorafenib in hepatocellular carcinoma.
我们旨在首先确定胡椒碱对细胞活力、细胞应激和细胞凋亡的影响，然后确定胡椒碱的作用与c-Jun氨基末端激酶（JNK）信号通路的关系，以及胡椒碱与索拉非尼在肝细胞癌中的相互作用。使用了肝细胞癌（HepG2和Hep3B）和非癌性肝细胞（AML12）细胞系。通过MTT法测定细胞活力。通过蛋白质免疫印迹法检测细胞应激、细胞凋亡和JNK信号标志物。细胞用SP600125作为JNK抑制剂进行预处理。使用CompuSyn软件计算胡椒碱的半数抑制浓度（IC50）值以及与索拉非尼的相互作用。胡椒碱对HepG2细胞的IC50值为97µM，对Hep3B细胞为58µM，对AML12细胞为184µM，孵育48小时。胡椒碱导致肝细胞癌细胞中的细胞应激、细胞凋亡显著增加，并激活JNK信号。用JNK抑制剂预处理可显著降低胡椒碱诱导的细胞应激、细胞凋亡和细胞毒性。在HepG2和Hep3B细胞中，胡椒碱与索拉非尼联合使用时具有浓度依赖性的相加或协同作用。我们发现胡椒碱通过JNK信号诱导细胞应激、细胞凋亡和细胞毒性，并且在肝细胞癌中与索拉非尼具有浓度依赖性的相加或协同作用。
