Induced Differentiation Of Hep G2 Human Hepatocarcinoma Cells And The Regulation Of Relative Cytokines By HCl-tetramethyl Pyrazine

objective:To investigate the effect of HCl-tetramethyl pyrazine on the proliferation, AFP secreation,mRNA and protein levels of TGF-βand VEGF of Hep G2 human hepatocarcinoma cells to evalule inductive effect on Hep G2 cell differentiation.Methods:The cell growth inhibition ratio was detect by MTT assay,and the cell morphology was watched by phase contrast microscope.The level of AFP protein in supernatant was detected by radio-immunological assay.The level of TGF-βand VEGF mRNA and protein were detected by Realtime-PCR and Western Blot to investigate the induced differentiation degree.ResuLts:1.HCl-tetramethyl pyrazine had certain extent of growth inhibition in contrast with the control and concentration dependent effect on the Hep G2 human hepatocarcinoma cells.The regression curve got the IC50 is 167.18μg/ml.2.HCl-tetramethyl pyrazine had some extent effect on cell morphology watched by phase contrast microscope,HCl-tetramethyl pyrazine with 1000μg/ml had an obvious cytotoxicity on Hep G2 human hepatocarcinoma cells,and the cell death happened in 48 hours.3.AFP protein was inhibited in certain degree with HCl-tetramethyl pyrazine treatment,and the miximum effect concentration is about 10μg/ml with significant difference with the control(P＜0.05).4.HCl-tetramethyl pyrazine had some extent effect on TGF-βand VEGF mRNA level.5.HCl-tetramethyl pyrazine had some extent effect on TGF-βand VEGF protein level.ConcLusions:HCl-tetramethyl pyrazine in low concentration has growth inhibition effect on the Hep G2 human hepatocarcinoma cells and some extent inhibition on AFP which is a hepatocarcinoma cell special protein,and TGF-βand VEGF mRNA and protein level were down-regulated.HCl-tetramethyl pyrazine in high concentration can inhibit cell growth and lead to cell death.So it is a potential cytotoxic medicine and tumor cells differentiation inducer.