Effect Of Wip1 Inhibitor On Invasion And Metastasis Of Intrahepatic Cholangiocarcinoma Cells

Background: Intrahepatic cholangiocarcinoma(ICC)is an epithelioid adenocarcinoma derived from the intrahepatic secondary bile duct and its branches.Its incidence is second only to liver cancer in primary liver tumors,and it is the second most common hepatic malignancy.Intrahepatic cholangiocarcinoma has no obvious clinical symptoms in the early stage,and early diagnosis is difficult.Most of the patients found with it are in the middle or late stage.Although some patients have the opportunity to get radical surgery,the recurrence rate is still very high,Which seriously restricts the patients' surgical outcome and prognosis,resulting in a total less than 10% rate for five-year survival.The previous studies in our unit found that the rate of Wip1 in ICC cases was as high as78.3%(47/60),but not in the adjacent cancer tissues,And in the meantime,the rate of Wip1 was higher in cases with lymphatic metastasis and neurological invasion.The difference is statistically significant.Related literatures at home and abroad report that Wip1 is highly-rated in a variety of human tumor cells,and its high rate therefore results in poor performances in prognosis.Objective: In summary,the early detection of intrahepatic cholangiocarcinoma and reduction of postoperative recurrence may greatly improve the survival time and prognosis of patients.Hence studying the invasion and metastasis mechanism of intrahepaticcholangiocarcinoma has great guiding significance for the treatment and prognosis of intrahepatic cholangiocarcinoma.Therefore,based on the previous studies of our unit,to investigate that inhibition of Wip1 rate can affect the proliferation and invasion ability of intrahepatic cholangiocarcinoma.Methods: First,the Wip1 specific inhibitor GSK2830371 was added to the intrahepatic cholangiocarcinoma cell line ICC-9810 in the experimental group.And then with the guidance of Western blot for the level of ICC-9810 Wip1 in the intrahepatic cholangiocarcinoma cells of the experimental group and the two control groups,and with CCK8,Transwell assay,and Scratch test,we are able to verify the changes in cell biological characteristics such as proliferation,migration and invasion of ICC-9810 cells.Results: Western blot analysis showed that the experimental group had lower grayscale values than the blank control group and the negative control group,the difference is statistically significant(p <0.05);The CCK8 experiment showed that the results of the third and fourth day were lower than those of the two control groups.The p value on the third day was less than 0.05,and the p value on the fourth day was less than 0.01;The Transwell assay showed that the number of cell invasions was significantly lower than that of the two control groups(p<0.05).The results of the Scratch test showed that the scratch widths of theexperimental group at 24 hours and the experimental group at 48 hours were significantly larger than those of the two control groups.The difference was statistically significant.(p<0.01).There were no significant differences between the two control groups in Western blot,CCK8,Transwell assay and Scratch test.Conclusion: GSK2830371 can reduce the level of Wip1 and inhibit the proliferation,invasion and metastasis of ICC-9810 cells.