Inhibitory Effect Of Mahong Pills Containing Serum On The Proliferation Of Human Hepatoma Carcinoma Smmc-7721 Cells And Its Mechanism

Objective: To investigate the inhibitory effect of Mahong pills containing serum on the proliferation of human hepatoma carcinoma smmc-7721 cells and its mechanism.Methods:Blank serum, Mahong pills and adriamycin (doxorubicin) containing serum were prepared by serologic pharmacological method.(1)MTT assay was used to obtain the proliferation inhibition rate of smmc-7721 cells after treated with blank serum, MHP and ADM containing serum; (2)The leakage of LDH in smmc-7721 cells of each group was determined, after treated with blank serum, MHP and ADM containing serum; (3)Nuclear morphological features of smmc-7721 cells were detected by fluorescence microscopy after staining with Hoechst33258; (4)Flow cytometry was used to detect the apoptotic ratio of smmc-7721 cells in each group; (5)The activity of telomerase of smmc-7721 cells in each group was determined by TRAP-PCR argentation method; (6) The change of bcl-2 and bax mRN A were dectected by RT-PCR.Results:(1)After treated with different volume of blank serum, ADM and MHP containing serum in 24,48 and 72 hours, the proliferation of smmc-7721 cells was significantly inhibited showing time and dose dependence, especially when the volume was 30% and the time was 72 hours, the inhibitory rate reached the highest(p<0.01);(2)The leakage of LDH significantly increased after treated with 30% MHP and ADM containing serum, compared to the cells in group of 30% blank serum(p<0.01);(3)The result of fluorescence microscopy observation showed condensation of cytoplasm and fragmentation of nucleus in cells of group treated with 30% MHP and ADM containing serum for 72 hours, and the apoptotic rate of cells obviously increased compared to cells in 30% blank serum group by flow cytometry method(p<0.01); (4)The result of TRAP-PCR argentation method showed that the activity of telomerase in cells of control and 30% blank serum group had no difference, but compared to 30% blank serum group, the activity of telomerase decreased in cells treated with 30% MHP and ADM containing serum for 72 hours;(5)RT-PCR analysis showed that the change of bcl-2 and bax mRNA in cells of control and 30% blank serum group had no difference, but compared to 30% blank serum group, bcl-2 mRNA in cells treated with 30% MHP and ADM containing serum for 72 hours was significantly down-regulated and bax mRNA was significantly up-regulated(p<0.01).Conclusion:(1)The proliferation of smmc-7721 cells can be significantly inhibited after treated with 30% Mahong pills containing serum for 72 hours;(2)30% Mahong pills containing serum can induce apoptosis of smmc-7721 cells;(3)The inhibitory effect of Mahong pills containing serum on smmc-7721 cells may be realized by suppressing the activity of telomerase, down-regulating bcl-2 mRNA, and up-regulating bax mRNA.