Preliminary Study On The Effect And Mechanism Of Triptonide Against Pancreatic Cancer Panc-1 Cells In Vitro

ObjectiveTo investigate the effect and mechanism of triptonide(Triptonide,TN)against human pancreatic cancer Panc-1 cells in vitro.Methods1.CCK-8(Cell Counting Kit-8)assay was used to detect the effect of different concentrations of TN on the proliferation of human pancreatic cancer Panc-1 cells in vitro.2.EdU cell proliferation test was used to detect the effect of TN on proliferation of Panc-1 cells.3.Colony formation test(Clonogenicity assay)was used to detect the effect of TN on colony formation of Panc-1 cells.4.Migration invasion assay(Transwell)was used to detect the effect of TN on the migration and invasion of Panc-1 cells.5.The effect of TN on apoptosis of Panc-1 cells was detected by flow cytometry.The expression of apoptosis-related proteins such as Cle-Caspase-3 was detected by Western blot.6.Comet assay was used to detect the effect of TN on DNA strand breaks in Panc-1 cells.7.Western blot was used to detect the expression of polynucleotide-5'phosphokinase-3' phosphatase(PNKP)in Panc-1 cells treated with TN.shRNA was used to inhibit the activity of PNKP in Panc-1 cells,and then the levels of PNKP and DNA strand breaks were detected by Western blot and comet assay respectively.8.shRNA was used to inhibit the activity of PNKP in Panc-1 cells,and then the colony formation and migration ability of Panc-1 cells were detected.Results1.TN could significantly inhibit the proliferation of Panc-1 cells at nM level,and the inhibitory effect increased with the increase of concentration.2.After TN treatment,the proliferation of Panc-1 cells was significantly decreased.3.After TN treatment,the colony forming ability of Panc-1 cells was significantly decreased.4.After TN treatment,the migration and invasion ability of Panc-1 cells was significantly decreased.5.TN could significantly induce apoptosis of Panc-1 cells.6.TN treatment resulted in an increase in DNA strand breaks in human Panc-1 cells.7.The expression of PNKP protein in Panc-1 cells decreased after TN treatment.After shRNA inhibited the expression of PNKP,DNA strand breaks increased in Panc-1 cells.8.After shRNA inhibited the expression of PNKP,the colony forming ability and migration ability of Panc-1 cells were significantly inhibited.Conclusion1.TN can significantly inhibit the proliferation,colony formation,weaken the ability of migration and invasion,and induce apoptosis of Panc-1 cells.2.The inhibitory effect of TN on Panc-1 cells may be related to the inhibition of PNKP expression.