Troxerutin with copper generates oxidative stress in cancer cells: Its possible chemotherapeutic mechanism against hepatocellular carcinoma

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. Cu is known to be elevated in cancer cells. Drug may have the possibility to interact with Cu and to alter homeostasis of cancer cells. In vitro study, shows that TXER + Cu complex exert significant Huh‐7 liver cancer cell death through apoptosis. Anticancer effect of TXER was confirmed in rat in vivo model. Our findings revealed the putative mechanisms behind the anticancer effect of TXER in hepatocellular carcinoma.