Inhibitive Effect Of Curcumol On Cholagiocarcinoma Cells And The Screening Of Drug Targets

Objective: Curcumol exerts strong anticancer effect but its role and specific target for cholangiocarcinoma is not yet clear.This paper intends to explore the role of curcumol in human cholangiocarcinoma cells,to study the effects of which on the biological characteristics of human cholangiocarcinoma cells(cell proliferation,migration,cell cycle and apoptosis)and to find effective therapeutic targets of curcumol.The potential target was validated by immunohistochemistry(IHC),and the further functional role in cholangiocarcinoma cells was detected by using Lentivirus-mediated vector.Methods: Human cholangiocarcinoma RBE and HCCC-9810 cell lines and cholangiocarcinoma tissues were used in this study.The inhibitory efficiency of curcumol on cholangiocarcinoma cells was estimated with CCK8 assay.Scratch assay was used to examine the cell migration ability.Alteration of cell cycle and apoptosis rate after curcumol treatments was detected by flow cytometry.The differentially expressed proteins of the curcumol-treated RBE cell proteins were gained based on the two-dimensional gel electrophoresis technology and verified by MALDI-TOF/TOF.CDKL3 was screened as a potential target of curcumol against cholangiocarcinoma and validated by using qRT-PCR and Western blot.We further investigated the role of silencing CDKL3 on the biological behavior of RBE cells by using RNA interference.The expression of CDKL3 in cholangiocarcinoma was validated by IHC and qRT-PCR.Result: Curcumol owns remarkably inhibitory effects on the proliferation and migration of cholangiocarcinoma cell lines and induces apoptosis via arrest cell cycle in G1 phase in a certain concentration range(50-100?g/ml)(P<0.01).Based on two-dimensional gel electrophoresis technology and bioinformatics We think of that CDKL3 might be as a potential target and which was used to the last experiments.qRT-PCR and Western blot data showed that the expression of CDKL3 was decreased after RBE cells treated with curcumol(P<0.01).After CDKL3 was down-regulated with lentiviral mediated sh RNA,the cell proliferation and migration ability were significantly reduced,and the proportion of G1 phase cells and the apoptosis rate were also increased(P<0.01).IHC and qRT-PCR data showed that CDKL3 was highly expressed in cholangiocarcinoma tissues.Conclusion:1.Curcumol had remarkably inhibitory effects on the proliferation and migration of cholangiocarcinoma cell lines and induce apoptosis via arrest cell cycle in G1 phase.CDKL3 was confirmed to be a potential target of curcumol for anti cholangiocarcinoma via t two-dimensional gel electrophoresis technology.The level of CDKL3 in the cholangiocarcinoma tissues is significantly elevated and may play a role in the oncogene.2.Down-regulation of CDKL3 could remarkably inhibit the proliferation and migration ability of cholangiocarcinoma cells and induce apoptosis via arrest cell cycle in G1 phase.Curcumol exerts anticancer effect in cholangiocarcinoma cells via down-regulating CDKL3...