JS-K,A Nitric Oxide Prodrug,Induces DNA Damage And Apoptosis In HBV-positive Hepatocellular Carcinoma HepG2.2.15 Cells And Safety Evaluation

Objective: This study aimed to examine anti-tumor effects of JS-K on HBV-positive hepatocellular carcinoma Hep G2.2.15 cells.Methods: The MTT assay and colony forming assay were used to study the cell growth inhibition of JS-K;scratch assay and transwell assay were performed to detect cell migration.The cell cycle was detected by flow cytometry.The immunofluorescence,flow cytometry analysis,and western blot were used to study DNA damage and cell apoptosis.Results: JS-K inhibited Hep G2.2.15 cell growth in a dose-dependent manner,suppressed cell colony formation and migration,arrested cells gather in the G2 phase,induce cell apoptosis.JS-K(1-20 ?M)increased the expression of DNA damage-associated protein phosphorylation H2AX(?H2AX),phosphorylation of checkpoint kinase 1(p-CHK1),phosphorylation of checkpoint kinase 2(p-CHK2),ataxia-telangiectasia mutated(ATM),phosphorylation of ataxia-telangiectasia mutated rad3-related(p-ATR)and apoptotic-associated proteins cleaved caspase-3,cleaved caspase-7,cleaved poly ADP-ribose polymerase(cleaved PARP).Conclusion: The study demonstrated JS-K is effective against HBV-positive Hep G2.2.15 cells,the mechanisms is related to induce DNA damage and apoptosis.JS-K is a promising anti-cancer candidate against HBV-positive HCC.Objective: This study aimed to perform preliminary safety evaluation on NO donor JS-K.Methods: To examine the long-term oral toxicity,male mice were given solvent control(CMC),JS-K low(2.5 mg/kg),middle(5 mg/kg)and high dose(10 mg/kg)orally for 3 months.The general health of animals and the body weights were recorded.At the end of experiments,the liver and kidney index,serum biochemistry and histopathology were examined.To examine the potential bone marrow toxicity,mice received intraperitoneal injection of JS-K(5 mg/kg)for 5 days,and the effects of JS-K on bone marrow and peripheral blood hematopoietic stem cells were determined via labelling with special antibodies followed by flow cytometry analysis.Results: The body weights were increased steadily with time in all groups,liver and kidney indexes and blood biochemistry parameters were all in the normal range.Hematoxylin and eosin stain showed no overt liver and kidney toxicity.The ratios of positive bone marrow stem cells of control were 2.19% and JS-K 1.98%.The ratios of positive peripheral blood hematopoietic stem cells in control were 0.11% and in JS-K 0.13%.The positive stem cells after injection with JS-K did not change.Conclusion: Oral JS-K for 3 months(up to 10 mg/kg)did not show overt toxicity to liver and kidney,injection JS-K for 5 days(5 mg/kg,4-fold of effective anticancer dose)did not affect bone marrow hematopoietic stem cells.