The Study On The Anti-tumor Effects Of Traditional Chinese Medicine Compound Liver Cancer No.1 For Hepatoma Carcinoma Cell

Objective: Chinese herbal medicine(CHM)is an effective aid for the clinical treatment of hepatocellular carcinoma cell(HCC),which is beneficial to the recovery of patients after partial hepatectomy and enhance the effect of molecular targeted drugs.The aim of this study was to investigate the anti-hepatoma effect and molecular mechanism of Chinese herbal compound liver cancer No.1.Methods: In vitro,MTT assay were detected cell proliferation of the normal liver cells HL-7702 and HCC SMMC-7721 treated with different concentrations of traditional Chinese medicine compound liver cancer No.1(1,2,4,8,12 and 16 mg/ml)for 24 h,48h and 72 h.Clone formation assay was detected cell proliferation of SMMC-7721 cells treated with different concentrations of traditional Chinese medicine compound liver cancer No.1(2,4,8 and 12 mg/ml)for 24 h.Flow cytometry assay was detected cell cycle arrest of different concentrations of traditional Chinese medicine compound liver cancer No.1(8 and 12 mg/ml)treated SMMC-7721 cells for 24 h.Flow cytometry apoptosis assay was detected cell apoptosis of different concentrations of traditional Chinese medicine compound liver cancer No.1(2,4,8,12mg/ml)treated with SMMC-7721 cells for 24 h.Wound healing assay was detected the ability of cell migration of different concentrations of traditional Chinese medicine compound liver cancer No.1(2,4 and 8 mg/ml)to treat with SMMC-7721 cells for 24 h.Transwell migration and invasion assay was used to detect the migration and invasion of SMMC-7721 cells treated with different concentrations of traditional Chinese medicine compound liver cancer No.1(2,4 and 8 mg/ml).Western blot assay was used to detect the difference concentration of Chinese medicine compound liver cancer No.1(2、4、8 and 12 mg/ml)treated with SMMC-7721 cells for 48 hours,the expression of proliferating cell nuclear antigen(PCNA),tumor necrosis factor receptor-associated factor 2(TRAF2),Caspase-8,Caspase-3,PARP,matrix metalloproteinase 2(MMP-2)and matrix metalloproteinase 9(MMP-9)protein.In vivo,the subcutaneous tumor xenografts in nude mice were used to determine whether the traditional Chinese medicine compound liver cancer No.1 could inhibit the growth of tumors.Results: The results of MTT assay showed that the traditional Chinese medicine compound liver cancer No.1 can inhibit the proliferation of HCC in a time and concentration dependent(p<0.05),but the inhibitory effect on normal liver was not obvious(p > 0.05).The results of clone formation assay showed that different concentrations of traditional Chinese medicine compound liver cancer No.1 can inhibit the proliferation of HCC with the increase of drug concentration,the less colonies fromation(p<0.05).The results of flow cytometric cycle showed that the traditional Chinese medicine compound liver cancer No.1 can block cells in the G2/M phase.The results of flow cytometry apoptosis showed that traditional Chinese medicine compound liver cancer No.1 significantly inhibited the early apoptosis and late apoptosis of cells(p<0.05).The results of wound healing showed that the traditional Chinese medicine compound liver cancer No.1 can inhibited the cell migration ability(p<0.05).Transwell migration and invasion assay results showed that the traditional Chinese medicine compound liver cancer No.1 significantly inhibited cell migration and invasion,and the number of migration cells were 149±12,105±19,68±9,and 21±4,the difference is statistically significant(p<0.05);the number of invasion cells were 89±9,61±4,35±9,and 17±5,the difference was statistically significant(p<0.01).The results of western blot assay showed that the expression of Cleaved Caspase-8,Cleaved Caspase-3,and Cleaved PARP protein was significantly up-regulated with the increase of traditional Chinese medicine compound liver cancer No.1;the expression of PCNA,TRAF2 of Full-length Caspase-8,Full-length Caspase-3,Full-length PARP,MMP-2 and MMP-9 proteins was significantly down-regulated.The results of tumor-bearing experiment in nude mice showed that traditional Chinese medicine compound liver cancer No.1 significantly inhibited the growth of tumors in vivo.Conclusions: The traditional Chinese medicine compound liver cancer No.1 can inhibit the proliferation of HCC,promote the apoptosis of HCC,reduce the migration and invasion ability of HCC,and also inhibit the growth of tumors in nude mice.