The Antitumor Mechanism Of Cryptotanshinone In Human HepG2 Cells

Cryptotanshinone is one of the effective components in salvia miltiorrhiza which has antitumor effect. In this thesis, its antitumor mechanism was studied with human HepG2 cell line as the test material through MTT, AO/EB double staining and RT-PCR. The results are as follows.1)Human HepG2 cell line was treated with cryptotanshinone of different concentrations for 6, 12, 18, 24 hours. Its OD value was assayed by MTT . It was found that the inhibiting effect of cryptotanshinone on human HepG2 cells treated for 24 hours were most obvious. OD values gradually decreased with the concentration of cryptotanshinone increasing, which meant that cryptotanshinone began to take its effect in a dose-dependent manner. The semi-inhibitory concentration of cryptotanshinone is 94.6μM : (IC50=94.6μM) .2)The apoptosis rate in HepG2 cells was detected through AO/EB double staining. The results of fluorescence detection showed that most cells were dyed with red while only a few cells were stained green, which meant that cryptotanshinone had the inhibiting effect obviously. It was deduced that cryptotanshinone activated the apoptosis pathway of HepG2 cells to inhibit the effect of HepG2.3)The expression of p53 and p21 gene was detected through semi-quantitative RT-PCR. Total RNA extracted from HepG2 cells treated with cryptotanshinone of different concentrations for 24 hours was used as templates for RT-PCR to detect mRNA expression of p53 and p21. The housekeeping gene GAPDH was used as internal standard. Through calculating target genes fragment and fragment of GAPDH content ratio, comparative concentration of the target genes in the total mRNA of HepG2 cells was obtained. Based on the experimental results, it was concluded that cryptotanshinone had obvious effect on p53 gene and no obvious effect on p21 gene.In this paper the anti-tumor mechanism of cryptotanshinone in HepG2 cells was studied. Cryptotanshinone had obvious effect on p53. It was infered that cryptotanshinone inhibited HepG2 cell through the activation of p53. Further study on HepG2 cell apoptosis is necessary. This research can provide new ideas and methods for producing new anti-tumor Chinese traditional medicine.