An Experimental Study On Induction Of Human Hepatocellular Carcinoma Cell Line Apoptosis By Tramets Robiniophila In Vitro

Objective To study the effect of Tramets robinophila on the apoptosis of the humanhepatocellular carcinoma cells line HepG-2 and understanding the mechanism of itstreatment for hepatocellular carcinoma. Methods MTT assay was used to test thetoxicity of the Chinese medicine Extractum trametes robiniophila murr and thechemosensitivity to chemotherapeutics in Chinese medicine Extractum trametesrobiniophila murr -treated HepG2 and HepG2/ADM cell lines. Flow cytometry wasused to test different doses of Extractum trametes robiniophila murr in the role ofapoptosis in HepG2 cells and apoptosis rate peaks; TUNEL method were used todetect the apoptotic rate by different concentration of Extractum trametes robiniophilamurr in HepG2 cells. DNA agarose electrophoresis were applied to analysis theappearance of DNA-LADDER with different concentrations of Extractum trametesrobiniophila murr treating HepG2 cells. Results The inhibition rates of HepG2 andHepG2/ADM caused by Chinese medicine Extractum trametes robiniophila murr wereless than 10% under the dose of 0.01g/L, and the IC50 were 0.917g/L and 1.66g/L.Chinese medicine Extractum trametes robiniophila murr(0.008g/L) partly overcame theMDR of cell line HepG2/ADM and decreased the drug resistance. The IC₅₀ of ADM,cisplatin (DDP), mytomycin (MMC), 5-fluororacil (5-FU) was descended obviously while with Chinese medicine Extractum trametes robiniophila murr(0.008g/L). Asmeasured by flow cytometry showed: the apoptosis rates(%) induced by Extractumtrametes robiniophila murr in concentration of group 0.1g/L,1g/L,5g/L,10g/Lwererespectively 3.6±0.53,7.25±0.96,8.96±1.56,13.65±1.38, the rates of negative controlgroup were 2.7±0.36, 5-FU group were 15.98±1.34, proved that apoptosis ratesincreased with the increasing of its concentration in HepG2 cells; apoptosis ratesinduced by the same concentration(5g/L) of Extractum trametes robiniophila murr inHepG2 cells in time of 24h,48h,72h were respectively 8.96±1.56,26.34±1.1,30.89±1.2, proved that the apoptosis rates also increased with the time of drugsextended. The results of detecting apoptosis by TUNEL methods showed: the apoptosisrates(%) induced by Extractum trametes robiniophila murr in concentration of group0.1g/L,1g/L,5g/L,10g/Lwere respectively 2.35±0.32,5.96±0.68,7.85±0.96,12.96±1.36, the rates of negative control group were1.23±0.21, and the rates of 5-FUgroup were 13.56±1.45. DNA agarose electrophoresis analysis showed that aftertreatment with different concentrations of Extractum trametes robiniophila murr HepG2cell DNA fragmentation turns into a difference of about 200 bp fragment. Apoptoticcells showed characteristics of the "ladder bands" (laddie). Conclusion Chinesemedicine Extractum trametes robiniophila murr can inhibit the growth of humanhepatocellular carcinoma in vitro. The possible mechanism is related to inhibit the DNAsynthesis and induce the apoptosis of hepatocellular carcinoma cell. It provide thetheory basis of Extractum trametes robiniophila murr for chemotherapy in clinic.