Glutamine Deprivation On Hypoxic Conditions In Hepatocarcinoma Cell HepG-2Expression Of HIF-1Effect

Objective: Hepatic arterial chemoembolization （TACE） as the preferred treatmentfor advanced hepatoma, its effect and status has been widely recognized, but thelong-term efficacy of poor. Research shows that, after transcatheter arterialchemoembolization in patients with tumor hypoxia is a leading cause of hepatomapostoperative recurrence and metastasis are the main factors, and hypoxia induciblefactor1alpha （HIF-1alpha） as a tumor hypoxia response key factor, in this processplays a crucial role in. To HIF-1a target for therapeutic intervention is becoming a kindof new anti tumor treatment strategy.In vivo extracellular glutamine as the most contentof amino acids, in tumor cell proliferation, metabolism play an important role in theprocess. Research suggests that glutamine deprivation inhibits lung cancer cellproliferation, metastasis, and can induce the expression of HIF-1reduction, and forhepatoma cells and the effects of no current research.The experimental application of CoCl₂chemical hypoxia method simulation oftumor hypoxia environment, CoCl₂as an iron chelator, can inhibit HIF-1proteindegradation, is a commonly used chemical hypoxia induced agent.The studies on glutamine deprivation of human hepatoma HepG-2cells in thehypoxic stimulation of cell cycle changes, HIF-1alpha expression changes of glutaminedeprivation index, as in hepatocellular carcinoma cells by HIF-1alpha as a target fortreatment strategies.Methods: Using CoCl₂chemical hypoxia method simulation of tumor hypoxiaenvironment, for hypoxia training; using flow cytometry （FCM） detection of glutaminedeprivation under hypoxic conditions HepG-2cell cycle changes; using RT-PCR andReal-Time-PCR method for the detection of glutamine deprivation on HepG-2cellsunder hypoxic conditions affect the expression of mRNA HIF-1alpha; Results:（1） Fluorescence microscope display, hypoxia after HepG-2cells relativeto control cells, cell size, cell membrane wrinkling, nucleoplasm ratio decreases,stacked into a group of HepG-2cell binding force decreased, loosely into individualfloating, scattered.Of glutamine deprivation and hypoxia treated cells demonstrated themore obvious.（2） Under hypoxic conditions HepG-2cells arrest in G1phase, whereasglutamine deprivation, hypoxia for G1phase of the cell cycle by block to reduce.（3）Glutamine deprivation after HIF-1alpha mRNA in hypoxic condition reducedexpression.Conclusion:（1） Glutamine deprivation inhibits hepatocellular carcinoma HepG-2cell viability, apoptosis induction;（2） Glutamine deprivation reduces hypoxicstimulation of HepG-2cells arrest in G1phase, thereby reducing the hypoxia response;（3） Glutamine deprivation at the level of transcription down-regulation of HIF-1alphaexpression.