The Effects Of Genistein On Human Pancreatic Cancer PANC-1cells

Objective:The effects of different concentrations of genistein on the study of the effect of PANC-1 on human pancreatic cancer cell line and expression of tumor suppressor genes,so as to provide new ideas for clinical treatment of pancreatic cancer.Methods:This research included experimental group(80,120,160 and 200μmol/L,Genistein),Standard control group(gemcitabine)and blank group(PBS).At the different timepoint(12,24,36,48 and 60h) after the cells of the three groups were disposed by different culture medium, Detect the proliferation activity of human pancreatic cancer cell line PANC-1 under the influence of Geniste through CCK-8 way and Test the percentage of the apoptotic cell under the effect of Genistein through the methed of FCM.Detect the express of tumor suppressor genes P53 and P21 through RT-PCR.Test the protein expression level of P53 and P21 by Western blot.The statistical analysis of apoptotic apoptotic and cell m RNA was the way of LSD. The statistical analysis of the proliferation activity was T test of paired Sample.Results:1. CCK-8 assay genistein on pancreatic cancer cells(PANC-1) growth inhibition Respectively in 160 μmol/L and 200 μmol/L, 120 μmol/L and 200 μmol/L, 160μmol/L and group GEM, 200 μmol/L and group GEM by SPSS 13 were analyzed by paired samples t test, the results showed no significant difference between the 160 μmol/L of the GEM group(P=0.067), while the remaining three set of P values were less than 0.05 for less than 36h; data, statistics found that 120 μmol/L is less than 0.05 of the GEM group of P value, and 200 μmol/L and GEM group showed no significant difference(P=0.051).Based on the above results, we think that genistein showed the overall concentration andtime dependent manner, the effect of 200μmol/L 36 h and GEM group is close to, but160μmol/L the overall effect is the best, and the GEM group is most nearly2. Each group was detected by flow cytometry in human pancreatic cancer cell line PANC-1 pro-apoptotic effects.FCM results can be seen, four genistein grouping, 80μmol / L of cell proliferation inhibition effect is not obvious, but the effect 120μmol / L and 160μmol / L closest to the GEM group, 200μmol / L late apoptotic cells accounted for a large section. Visible effects of genistein have significant concentration-dependent manner.3. Protein expression levels of Western-blot detection of P21, P53 in each drug group Western-blot detection of protein levels show 160μmol / L and 200μmol / L genistein relative to the control can significantly enhance the expression of the tumor suppressor protein P21 and P53 gene, no significant differences with respect to the standard group.Conclusions:1.By flow cytometry and by CCK-8 genistein showed a concentration-dependent and time-dependent manner, but at low concentrations, the role of genistein is not ideal, but when the concentration of 160μmol/L or more of its FCM when the results of the control group with standard gemcitabine is very close to the concentration of 200μmol/L late apoptotic cells accounted for more than 60%.2.CCK-8 law proliferation display 160μmol/L at 48 h inhibitory effect to achieve the best, the best and the overall effect is similar to the curve with the standard controls, and Western-blot showed that genistein on P21, P53 protein were significantly increased the role of a small concentration-dependent.3.We conclude that the concentration of 160μmol/L genistein on pancreatic cancer cell line PANC-1 inhibition of the best, it may pass down cell proliferation, cell cycle-related genes and raised P21, P53 protein, so that genistein in pancreatic cancer inhibition of cell line PANC-1 proliferation showed concentration and time-dependent manner, and thus play a role in inhibiting cancer.