Studies On The Mechanism Of The Induction Of Apoptosis In Hepatocell?lar Carcinoma Cells By A Novel Indolyl Chalcone Derivative

Liver cancer is one of the most common cancers in the world and the third most common cause of cancer death.The prevalence of liver cancer is high in the world,especially in developing countries such as East Asia and South Africa.The liver is an organ that is susceptible to oxidative damage.When ROS is overproduced,the homeostasis will be disturbed,leading to oxidative stress.Oxidative stress plays a crucial role in the occurrence and development of liver diseases.High levels of oxidative stress result in the oxidative modification of amino acid residues,thereby inducing DNA mutations and apoptosis.Apoptosis is widely considered to be the most common form of liver injury.Intracellular mitochondrial metabolic disorder or reactive oxygen species(ROS)excessively cause mitochondrial damage,leading to cytochrome C release and caspase-3 activation,thereby activating the caspase cascade.At present,the main treatment methods for liver cancer include physical therapy and chemotherapy.During the course of treatment,patients suffer from a variety of adverse reactions and decreased tolerability,and there is an urgent need for new,less toxic therapeutic agents for the treatment of liver cancer.Chemotherapeutic drugs play an important role in anti-tumor by triggering the killing of cancer cell apoptosis.Therefore,the discovery and research of new therapeutic targets provide a new and reasonable method for the treatment of liver cancer.As an anti-cancer drug,small molecules have the characteristics of overcoming drug resistance,improve the prognosis and survival of cancer patients,and they can be used as new treatments for cancer.Many studies have shown that mercapto derivatives and chalcone derivatives have anti-tumor and apoptosis promoting effects.From this,we speculate that decylchrysone derivatives may also be an anti-cancer drug.In this study,a series of fluorenyl chalcones derivatives were synthesized by the College of Chemistry of Jilin University for in vitro cytotoxicity screening.We found that the screened compound 5h2 c has an effective cytotoxicity on hepatoma cells.Subsequently,we further validated 5h2 c inhibition of HepG2 and PLC/PRF /5 cell proliferation by MTT cell proliferation assay.The mitochondrial membrane potential was detected by flow cytometry and the intracellular reactive oxygen species was detected by DCFH-DA fluorescence probe.The results showed that 5h2 c could change mitochondrial membrane potential and induce ROS production.In addition,In addition,flow cytometry was used to detect cell cycle distribution and apoptosis.The results confirmed that 5h2 c can cause G0/G1 phase arrest and promote apoptosis in hepatoma cells.At the same time,we found from Transwell experiments and wound healing experiments that 5h2 c could inhibit the migration of HepG2 and PLC/PRF/5 cells.We then performed apoptotic array kit assays to search for molecular targets of apoptosis in HCC cells.The results showed that 5h2 c caused differential expression of multiple proteins.Bcl-2 family proteins and heat shock proteins are the two proteins that are most dramatically induced by 5h2 c.We analyzed Western blot results and observed that 5h2 c can increase the Bax/Bcl-2 ratio,inhibit the expression of heat shock proteins,and trigger mitochondrial and death receptor-mediated apoptosis.In addition,we also established a xenograft model in nude mice and confirmed that 5h2 c inhibits HCC tumor progression in vivo.At the same time,we found that 5h2 c treatment did not cause significant body weight changes in nude mice and did not produce side effects.In summary,5h2 c may be a potential anticancer drug candidate for the treatment of liver cancer.