Inhibition Icaritin Combined With Fluorouracil On Human Hepatoma HepG2 Cell Proliferation

Objective: Human HepG2 cells for research material, combined with fluorouracil icaritin investigate inhibition of human hepatoma HepG2 cell proliferation and inhibition of migration and restoration, as icaritin combined with fluorouracil as an antitumor drug combination provides a theoretical basis.Methods:In vitro treatment with different drugs HepG2 cells, the cells were observed under an inverted microscope adherent conditions and morphological changes, cell scratch with tips do observe restoration and migration of HepG2 cells after treatment with different drugs in different time periods Different role in drug use after a period of time blue tetrazolium (MTT) assay of cell growth inhibition, and calculate its growth inhibition rate (IR), and drug interaction coefficient (CDI), with flow cytometry analysis of apoptosis after treatment with different drugs, different drug treatment analysis of the cell cycle by flow cytometry.Statistical method for processing: All data applications SPSS 18.0 statistical analysis, measurement data described by the mean ± standard deviation (x±s), the single factor between the groups treated samples were compared using ANOVA number of multiple and LSD-t test, the two groups were compared using a t test. Test level α= 0.05, the definition of P<0.05 considered statistically significant.Results:1 under an inverted microscope growth state of the cells found in normal HepG2 well clear cell outline.5-FU group, ICT group and ICT + 5-FU group with increasing duration of action of the drug, the cells gradually become irregular, the number of shrinking, growing filopodia, cytoplasmic vacuoles, adherent cells gradually poor, emerging apoptotic bodies, cell death increased gradually shedding, increased cell debris.2. MTT find colorimetric assay ICT,5-FU, ICT+ 5-FU groups act on human hepatoma HepG2 cells significantly inhibited proliferation in a time-dependent manner. But also found in the 10～80umol/1 concentration range of ICT alone and combined with a concentration-dependent inhibition was also found 5-FU on human hepatoma HepG2 cell proliferation; different concentrations of 5-Fu ICT and joint use of human hepatoma HepG2 cells Inhibition of proliferation compared with the respective monotherapy was significantly increased (P<0.05). ICT (10umol/1, 20umol/1,40umol/1) and the combined effects of 5-FU in human hepatoma HepG2 cells 24h,48h,72h, two drugs have a synergistic effect (CDI<1), and ICT (80umol/1) and 5-FU combined effects of human hepatoma HepG2 cells 48h,72h, two antagonistic drugs (CDI> 1).3. The results of flow cytometry cycle 5-FU, ICT are on the S phase of the cell cycle arrest in human hepatoma HepG2 role, and found after the combination of S-phase cell cycle arrest in HepG2 and respective monotherapy group its blockade enhanced.4. Flow cytometry apoptosis results can be seen in each group of drugs on human hepatoma HepG2 cells were inhibited. Wherein the number of the combined group and the respective single drug group on human hepatoma HepG2 cell apoptosis was significantly increased. Each group of drug-induced human hepatoma HepG2 cell apoptosis drug trends and the results are generally consistent with MTT.5. cell scratch was found in each group of drugs on HepG2 cells were significantly inhibited migration and the role of repair, with the extension of time for each group of drug action, its role is more obvious. Where the combined group with the corresponding single-drug group on human hepatoma HepG2 cell migration and repair inhibition was significantly enhanced.Conclusion:icaritin on human hepatoma HepG2 cell proliferation was inhibited in a time - a concentration-dependent manner, combined with 5-FU after its inhibitory effect is more significant. Icaritin on human hepatoma HepG2 cells in S phase cycle arrest, combined with 5-FU after its enhanced role in S phase cycle arrest. Icaritin on human hepatoma HepG2 cells can promote apoptosis, combined with 5-FU after his promotion of apoptosis is enhanced. Icaritin, fluorouracil on human hepatoma HepG2 cells have the ability to inhibit the migration and repair, joint 5-FU after their migration inhibitory capacity enhancement and restoration.