Effect of Nux Vomica on growth of hepatocellular carcinoma cells and action mechanism

Abstract

Objective To investigate the inhibitory effects of Nux Vomica on human hepatocellular carcinoma (HCC) cells and the action mechanism.Methods Human HCC SMMC-7721 cells were cultured and the serum from Nux Vomica at the concentrations of 5％,10％,20％ was added into cell culture fluid respectively.The inhibitory effect of Nux vomica on SMMC-7721 cells was measured by using methyl thiazol tetrazolium (MTT) assay.After treatment of SMMC-7721 cells with the serum from 20％ Nux Vomica for 0,24 and 48 h,the cells were collected,and the total protein and RNA were extracted.The protein and mRNA expression of Fas,Cyclin D1 and PCNA was detected by using Western blotting and fluorescence quantitative polymerase chain reaction (PCR) method.Results The proliferation of SMMC-7721 cells was obviously inhibited by the serum from Nux Vomica in a concentration-dependent manner.The inhibition ratio was (54.94 ±8.19)％ with the serum from 20％ Nux Vomica and there was statistically significant difference in comparison with the serum from 5％ and 10％ Nux Vomica [ ( 19.928 ±7.653)％,(29.020 ± 6.100) ％ ] ( F =18.23,P ＜ 0.01 ).The relative expression of Fas protein was respectively (0.302 ±0.009) and (0.399 ±0.021 ) at 24 and 48 h respectively,which was significantly higher than in control group ( 0.226 ± 0.022) ( F =68.25,P ＜ 0.05 ).The relative expression of PCNA protein was 0.7457 ±0.0386 at 48 h,which was significantly lower than in control group and 24 h group (0.8529 ±0.0099,0.9016 ±0.0139) ( P ＜ 0.05 ).The Cyclin D1 mRNA expression was 0.045680 ± 0.038130 and 0.026714 ± 0.019934 at 24 and 48 h respectively,which was significantly difference from that in control group (0.145 246±0.048 543,F=8.671,P ＜0.05).There was no significant difference in the cyclinD1 protein expression ( F =68.25,P ＞ 0.05 ).Conclusion The proliferation of HCC cells can be inhibited with Nux Vomica in a dose-dependent manner.The high expression of Fas,low expression of PCNA as well as low expression of Cyclin D1 mRNA in HCC cells is closely correlated to the inhibitive effect on tumor cell proliferation. Key words: Nux vomica; Carcinoma,hepatocellular; Mechanism