Cyclophosphamide with Adjuvant Chemotherapeutic Drugs Induces Epigenetic Changes in Hepatocellular Carcinoma Cells

摘要

Cancer remains the major health threat worldwide; therefore, the extensive search for potent cancer-controlling agents are still a big demand. Hepatocellular carcinoma (HCC) is a type of cancer widespread in the developing countries. In the present study, the role of Cyclophosphamide and drug combinations (including Erlotinib, Temozolomide, Vorinostat, and Sodium Phenylbutyrate) as DNA methyltransferase (DNMT) and Histone deacetylase (HDAC) inhibitors was evaluated. Two concentrations of each drug i.e ., 3μM and 5μM for one incubation period of 72 h were applied. Trypan blue test was used to count the number of viable cells before and after treatments. DNA degradation assay was employed to evaluate the effect of Cyclophosphamide and a combination of drugs on the integrity of genomic DNA. Global methylation was also quantified via measuring the concentration of 5-Methylcytidin in the treated and un-treated HCC cells. Data obtained indicated that treating HCC cells with Cyclophosphamide either alone or in combination with other drugs has resulted in a significant decrease in the number of viable cells. Meanwhile, global DNA methylation data analysis showed that three combinations have resulted in hypomethylating the whole genome of HCC cells (Cyclophosphamide with Erlotinib, Cyclophosphamide with Sodium Phenylbutyrate, and Cyclophosphamide with Vorinostat). Although in vitro data need to be tested on the pre-clinical level, the best combination, Cyclophosphamide combined with Sodium Phenylbutyrate, might be recommended to be used in treating HCC in vivo .