Subastri Ariraman, Suyavaran Arumugam, Preedia Babu Ezhuthupurakkal, Nithyananthan Subramaniyam, Barathidasan Rajamani, Thirunavukkarasu Chinnasamy
Department of Biochemistry and Molecular Biology, Pondicherry University, Puducherry, India.
Centre for Animal Research, Training and Services, CIDRF-DBT, Sri Balaji Vidyapeeth University, Puducherry, India.
J Cell Physiol. 2018 Mar;233(3):1775-1790. doi: 10.1002/jcp.26061. Epub 2017 Aug 3.
Troxerutin (TXER) a rutin derivative is known for its anticancer effect against hepatocellular carcinoma (HCC). As part of large study, recently we have shown TXER interact with genetic material and its anti-mutagenic property. In the present study we have explored its possible mode of action in HCC. Since TXER alone did not show significant anticancer effect on Huh-7 cells, in vitro biochemical assays were performed for determining anticancer efficacy of TXER + metal complex using transition metals such as Cu, Zn, and Fe. The anticancer efficacy of TXER + Cu on Huh-7 cells were evaluated using MTT assay, DCFDA, JC-1 staining, comet assay, cell cycle analysis, immunocytochemistry, and Western blotting. Non-toxic nature of TXER was analyzed on primary rat hepatocytes. The in vivo efficacy of TXER was tested in N-nitrosodiethylamine initiated and γ-benzene hexachloride and partial hepatectomy promoted rat liver cancer. Liver markers, transition metal levels, histopathological examination, and expression levels of GST-P, 8-OHdG and Ki-67 were studied to assess the in vivo anticancer effect of TXER. We observed that TXER + Cu induced extensive cellular death on Huh-7 cells through generating free radicals and did not possess any toxic effect on normal hepatocytes. The in vivo studies revealed that TXER possess significant anti-cancer effect as assessed through improved liver markers and suppressed GST-P, 8-OHdG, and Ki-67 expression. TXER treatment reduced the hepatic Cu level in cancer bearing animals. Current study brings the putative mechanism involved in anti-cancer effect of TXER, further it will help to formulate phytoconstituents coupled anti-cancer drug for effective treatment of HCC.
曲克芦丁（TXER）是一种芦丁衍生物，以其对肝细胞癌（HCC）的抗癌作用而闻名。作为一项大型研究的一部分，最近我们发现TXER与遗传物质相互作用及其抗诱变特性。在本研究中，我们探讨了其在HCC中可能的作用方式。由于TXER单独对Huh-7细胞未显示出显著的抗癌作用，因此使用铜、锌和铁等过渡金属进行体外生化测定，以确定TXER +金属络合物的抗癌功效。使用MTT法、DCFDA、JC-1染色、彗星试验、细胞周期分析、免疫细胞化学和蛋白质印迹法评估TXER +铜对Huh-7细胞的抗癌功效。在原代大鼠肝细胞上分析TXER的无毒性质。在N-亚硝基二乙胺引发、γ-六六六和部分肝切除术促进的大鼠肝癌模型中测试TXER的体内功效。研究肝脏标志物、过渡金属水平、组织病理学检查以及GST-P、8-羟基脱氧鸟苷（8-OHdG）和Ki-67的表达水平，以评估TXER的体内抗癌作用。我们观察到TXER +铜通过产生自由基诱导Huh-7细胞大量死亡，并且对正常肝细胞没有任何毒性作用。体内研究表明，通过改善肝脏标志物以及抑制GST-P、8-OHdG和Ki-67的表达评估，TXER具有显著的抗癌作用。TXER治疗降低了荷瘤动物肝脏中的铜水平。当前研究揭示了TXER抗癌作用的推定机制，进一步将有助于配制植物成分联合抗癌药物以有效治疗HCC。
