Bondada Venkata Mani Anirudh1, Devaraj Ezhilarasan2,3. 1. Department of Pharmacology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences (SIMATS), No. 162, PH Road, Chennai, Tamil Nadu, 600077, India. 2. Department of Pharmacology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences (SIMATS), No. 162, PH Road, Chennai, Tamil Nadu, 600077, India. ezhild@gmail.com. 3. Biomedical Research Unit and Laboratory Animal Centre, Saveetha Dental College (SDC), Saveetha Institute of Medical and Technical Sciences, Chennai, Tamil Nadu, 600077, India. ezhild@gmail.com.
Abstract
BACKGROUND: Colorectal cancer (CRC) is one of the frequently diagnosed cancers worldwide. Currently used chemotherapeutic drugs have several side effects. Histone deacetylase (HDAC) enzyme inhibitors possess potential anti-cancer effects. Therefore, we investigated the cytotoxic potential of sodium valproate, a HDAC inhibitor in human colorectal adenocarcinoma (HT-29) cells. METHODS: MTT assay was used to analyze the cytotoxicity of HT-29 cells. Intracellular reactive oxygen species (ROS) induction was evaluated by dichloro-dihydro-fluorescein diacetate staining. Dual staining with acridine orange/ethidium bromide was used to investigate the morphology-related apoptotic cell death. Mitochondrial membrane potential was analyzed by rhodamine 123 staining. E-cadherin protein expression was examined by immunofluorescence staining. RESULTS: Sodium valproate at 2 and 4 mM/mL treatments significantly induced cytotoxicity. Increased intracellular ROS expression was observed in the cells treated with sodium valproate. This treatment also induced mitochondrial dissipation, apoptosis-related morphological damage, and E-cadherin expression in HT-29 cells. CONCLUSIONS: Our present results suggest that sodium valproate is cytotoxic to HT-29 cells due to its pro-oxidative and apoptosis inducing potential. Sodium valproate can be used as an adjuvant along with standard chemotherapeutic agents in CRC patients after necessary in vivo and clinical studies.
BACKGROUND: Colorectal cancer (CRC) is one of the frequently diagnosed cancers worldwide. Currently used chemotherapeutic drugs have several side effects. Histone deacetylase (HDAC) enzyme inhibitors possess potential anti-cancer effects. Therefore, we investigated the cytotoxic potential of sodium valproate, a HDAC inhibitor in human colorectal adenocarcinoma (HT-29) cells. METHODS: MTT assay was used to analyze the cytotoxicity of HT-29 cells. Intracellular reactive oxygen species (ROS) induction was evaluated by dichloro-dihydro-fluorescein diacetate staining. Dual staining with acridine orange/ethidium bromide was used to investigate the morphology-related apoptotic cell death. Mitochondrial membrane potential was analyzed by rhodamine 123 staining. E-cadherin protein expression was examined by immunofluorescence staining. RESULTS: Sodium valproate at 2 and 4 mM/mL treatments significantly induced cytotoxicity. Increased intracellular ROS expression was observed in the cells treated with sodium valproate. This treatment also induced mitochondrial dissipation, apoptosis-related morphological damage, and E-cadherin expression in HT-29 cells. CONCLUSIONS: Our present results suggest that sodium valproate is cytotoxic to HT-29 cells due to its pro-oxidative and apoptosis inducing potential. Sodium valproate can be used as an adjuvant along with standard chemotherapeutic agents in CRC patients after necessary in vivo and clinical studies.