Screening And Anti-tumor Activity Of PLK1 PBD Targeted Inhibitors

Objective:In order to find lead compound which is capable,at least in vitro,of inhibiting tumor cells’ growth by restraining PLK1 PBD’s activity.Methods:Protein purification by PLK1 protein purification techniques.We used fluorescence polarization model to screen positive compounds targeting PLK1 PBD.MTT assay was utilized to find the most sensitive cell lines of 1097C11.The effects of the positive compound on cell cycle and apoptosis were all tested by flow cytometry(FCM).The combination of compound with PLK1 PBD was determined by molecular docking.Compound’s effect on cell movement was determined by cell migration experiment.Results:We obtained PLK1 PBD recombinant protein concentration of 0.723mg/mL by protein purification techniques.We used the fluorescence polarization model for high-throughput screening from about 40000 compounds.Directional obtain new structure lead compounds 1097C11 and 6509G11 which can inhibit the activity of PLK1 PBD.Through MTT we found that 6507 G11 and 1097 Cll has obvious cytotoxicity on 8 strains of tumor cell and the IC50 of HCT-116 are(5.38±0.16)μM and(8.43±0.58)μM.In the cell cycle block analysis found that 6507G11 and 1097 C11 could clearly make cycle synchronization HCT-116 cells G2/M phase retardation by dose dependence to delay cell cycle process.15μM 6507G11 can made 90.61%late apoptotic cells from the total cells.25μM 1097C11 can made 77.02%late apoptotic cells from the total cells.In the Annexin V/PI double dye we found that 6507 G11 and 1097 C11 can obviously dose dependent make HCT-116 cells apoptosis.Cell migration experiment shows that 6507G11 and 1097C11 can inhibit HCT-116 cell migration ability,and presents the time and the concentration dependencee molecular docking virtual results showed that the 6507G11 and 1097C11 have good affinity in structure domain with PLK1 PBD.Conclusion:Get a higher degree of purification of the protein PLK1 PBD 6507G11 and 1097C11 which has good antitumor activity is expected to be a lead compound targeting PLK1 relevant tumor.