Samie Nima, Muniandy Sekaran, Kanthimathi M S, Haerian Batoul Sadat
Department of Pharmacology, Faculty of Medicine, University of Malaya , Kuala Lumpur , Malaysia.
Department of Molecular Medicine, Faculty of Medicine, University of Malaya , Kuala Lumpur , Malaysia.
PeerJ. 2016 Mar 17;4:e1588. doi: 10.7717/peerj.1588. eCollection 2016.
The purpose of this study was to assess the cytotoxic potential of a novel piperazine derivative (PCC) against human liver cancer cells. SNU-475 and 423 human liver cancer cell lines were used to determine the IC50 of PCC using the standard MTT assay. PCC displayed a strong suppressive effect on liver cancer cells with an IC50 value of 6.98 ± 0.11 µM and 7.76 ± 0.45 µM against SNU-475 and SNU-423 respectively after 24 h of treatment. Significant dipping in the mitochondrial membrane potential and elevation in the released of cytochrome c from the mitochondria indicated the induction of the intrinsic apoptosis pathway by PCC. Activation of this pathway was further evidenced by significant activation of caspase 3/7 and 9. PCC was also shown to activate the extrinsic pathways of apoptosis via activation of caspase-8 which is linked to the suppression of NF-κB translocation to the nucleus. Cell cycle arrest in the G1 phase was confirmed by flow cytometry and up-regulation of glutathione reductase expression was quantified by qPCR. Results of this study suggest that PCC is a potent anti-cancer agent inducing both intrinsic and extrinsic pathways of apoptosis in liver cancer cell lines.
本研究的目的是评估一种新型哌嗪衍生物（PCC）对人肝癌细胞的细胞毒性潜力。使用SNU - 475和423人肝癌细胞系，通过标准MTT试验确定PCC的半数抑制浓度（IC50）。处理24小时后，PCC对肝癌细胞显示出强烈的抑制作用，对SNU - 475和SNU - 423的IC50值分别为6.98±0.11µM和7.76±0.45µM。线粒体膜电位显著下降以及线粒体中细胞色素c释放增加，表明PCC诱导了内源性凋亡途径。半胱天冬酶3/7和9的显著激活进一步证明了该途径的激活。PCC还通过激活与抑制NF-κB向细胞核转位相关的半胱天冬酶-8，显示出激活凋亡的外源性途径。通过流式细胞术确认细胞周期停滞在G1期，并通过定量聚合酶链反应（qPCR）对谷胱甘肽还原酶表达的上调进行定量。本研究结果表明，PCC是一种有效的抗癌剂，可诱导肝癌细胞系内源性和外源性凋亡途径。
