Study On The Effect And Mechanism Of The Chinese Herbs Of Insect Protein Extract With Liver Channel Tropism In Human Heaptoma HepG2 Cells

Objective:To analyse the mechanisms and inhibition effects of four Chinese herbs of insect with liver channel tropism scorpion protein extract(SPE), centipede protein extract (CPE), eupolyphaga sinensis walker protein extract (EPE), leech protein extract (LPE), to HepG2 cell proliferation.Methods:All aqueous extracts, the SPE, CPE, EPE, LPE were prepared by methods of rapid freezen-thaw with liquid nitrogen, and following precipitation with acetone. The proliferation inhibition rates of four drugs were assayed by MTT method. Cell cycle were detected by flow cytometry. The HepG2 cell apoptosis were detected by AO/EB fluorescent staining. Oncogene-C-myc, N-ras and untioncogene-PTEN, p53 mRNA expression were detected by RT-PCR and Immunohistochemistry.Results:1. Four Chinese herbs of insect with liver channel tropism- scorpion protein extract(SPE), centipede protein extract (CPE), eupolyphaga sinensis walker protein extract (EPE), leech protein extract (LPE) are all dose dependent, that is, as the concentration of chemical increase, the effects will also increase. In which CPE has the strongest effect, and EPE had no significant effect.2.The IC50 are SPE 7.02mg/ml, CPE 3.07mg/ml, LPE 9.83mg/ml, EPE 433.57mg/ml respectively.3. Under the effects of IC50 concentration, the effects are not significant; the comparative difference with control group has no statistical significance (P>0.05); the effects at 48h and 72h is significant; the four Chinese herbs of insect can inhibit the proliferation; the comparative difference with control group has statistical significance (P<0.01)4. the effects of all extracts at 24h-compared to the negative control group, SEP and CPE can significantly cause cell apoptosis significantly (P< 0.01), while EPE and LPE cannot (P>0.05). The effects of all extracts at 48h-compared to the control group, SEP, CPE, EPE and LPE can cause cell apoptosis, which has statistical significance (P<0.01)5. Compared to the negative control group, under the effects of SPE, CPE, LPE the cell cycle representation shows that the cell proportion decrease at G0/G1 period and increase at S period and G2/M, which has statistical significance (P<0.01) Compared to the negative control group, under the effects of EPE, the cell proportion decrease at G0/G1 period and increase at S period and G2/M, which has statistical significance (P <0.01).6. Compared to the control group, SEP can down regulate the N-ras mRNA and up regulate the PTEN mRNA, which has statistical significance (P<0.01); Compared to the control group, CPE can down regulate the C-myc, N-ras mRNA and up regulate the PTEN mRNA, which has statistical significance (P<0.01); Compared to the control group, EPE can down regulate the C-myc mRNA, which has statistical significance (P<0.01); Compared to the control group, LPE can down regulate the C-myc, N-ras and p53 mRNAN, which has statistical significance (P<0.01). Conclusions:1. Theory as the guide to go through the screening with anti-hepatoma effect of traditional Chinese medicine active ingredients feasible.2. Chinese Herbs of Insect with Liver Channel scorpion,centipede,eupolyphaga sinensis walker,leech protein extracts from the role of anti-hepatoma HepG2 cells, there were differences between drug reduced mass owned by attacking the liver of the Scorpio, centipedes anti-liver cancer is superior to anticoagulant, the Eupolyphaga pass through liver worms, leeches.3. Chinese herbs of insect with liver channel regulation the representation of cancer-related genes C-myc,N-ras,p53,PTEN mRNA, Thereby inhibiting the proliferation of hepatoma cells or apoptosis, it may go to the liver by the insects an important traditional Chinese medicine molecular mechanism of anti-liver cancer.