Endah Endah, Wulandari Febri, Putri Yurananda, Jenie Riris Istighfari, Meiyanto Edy
Department of Biotechnology, Graduate School, Universitas Gadjah Mada, Yogyakarta, Indonesia.
Cancer Chemoprevention Research Center, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia.
Iran J Pharm Res. 2022 Feb 4;21(1):e123820. doi: 10.5812/ijpr.123820. eCollection 2022 Dec.
Pentagamavunon-1 performs more potent anti-cancer effects than curcumin against various cancer cells, but it remains to be optimized. Piperine shows the activity as an enhancer of a therapeutic agent. This study expects to achieve higher effectiveness of PGV-1 on 4T1 breast cancer cells through co-treatment with piperine with exploring the effect of cytotoxicity, mitotic catastrophe, cellular senescence, and target proteins of PGV-1 and piperine on the regulation of mitosis in TNBC cells (4T1). The assays emphasize MTT assay, staining, SA-β-galactosidase assay, and bioinformatics analysis, respectively, to elicit the respected activities. The results revealed that PGV-1 performed a cytotoxic effect with an IC50 value of 9 µM while piperine showed a lower cytotoxic effect with an IC50 value of 800 µM on 4T1 cells 24 h treatment. However, the combination treatment of both showed a synergistic cytotoxic enhancement effect with an average CI value < 1. Furthermore, the combination of PGV-1 and piperine induced mitotic catastrophe and senescence better than the single treatment. Treatment of 1 µM of PGV-1 and 400 µM of piperine increased the percentage of senescent cells by 33%. Bioinformatics analysis revealed that PGV-1 and piperine target proteins play a role in mitotic regulation, namely CDK1, KIF11, AURKA, AURKB, and PLK1, to contribute to mitotic catastrophe. Therefore, piperine increases the effectiveness of PGV-1 to suppress 4T1 cells growth synergistically that may occur through mitotic catastrophe and senescence targeting on mitotic regulatory proteins.
五聚半胱氨酸-1（Pentagamavunon-1）对多种癌细胞的抗癌作用比姜黄素更强，但仍有待优化。胡椒碱具有作为治疗剂增强剂的活性。本研究期望通过与胡椒碱联合处理，在探索细胞毒性、有丝分裂灾难、细胞衰老以及PGV-1和胡椒碱的靶蛋白对三阴性乳腺癌细胞（4T1）有丝分裂调节作用的基础上，提高PGV-1对4T1乳腺癌细胞的疗效。这些实验分别着重于MTT实验、染色、SA-β-半乳糖苷酶实验和生物信息学分析，以得出相应的活性。结果显示，在对4T1细胞进行24小时处理时，PGV-1表现出细胞毒性作用，IC50值为9µM，而胡椒碱的细胞毒性作用较低，IC50值为800µM。然而，两者的联合处理显示出协同的细胞毒性增强作用，平均CI值<1。此外，PGV-1和胡椒碱的联合处理比单一处理能更好地诱导有丝分裂灾难和衰老。用1µM的PGV-1和400µM的胡椒碱处理可使衰老细胞的百分比增加33%。生物信息学分析表明，PGV-1和胡椒碱的靶蛋白在有丝分裂调节中发挥作用，即细胞周期蛋白依赖性激酶1（CDK1）、驱动蛋白家族成员11（KIF11）、极光激酶A（AURKA）、极光激酶B（AURKB）和 polo样激酶1（PLK1），从而导致有丝分裂灾难。因此，胡椒碱可协同提高PGV-1抑制4T1细胞生长的有效性，这可能是通过针对有丝分裂调节蛋白的有丝分裂灾难和衰老实现的。
