Effects Of Propofol On Metastasis And EMT Of Human Hepatocellular Carcinoma Cells Through Wnt/β-catenin Signaling Pathway

Background and Objective:Hepatocellular carcinoma(HCC)is one of the most common malignancies in China.Its high rates of recurrence and metastasis have become a major obstacle to the prognosis of HCC patients.Surgical resection and liver transplantation is the most important method of treating liver cancer,but the prominent problem of surgical resection is high recurrence rate.The current study shows that surgical operation and stress can promote the growth and invasion of tumor cells,perioperative period is the prognosis of patients with recurrence and long-term prognosis of one of the factors.Therefore,the anesthesiologist in the liver malignant tumor perioperative use of inhibition of liver cancer cell growth and invasive anesthetic drugs or methods,theoretically to maximize the avoidance of liver cancer cells during the perioperative spread.Propofol is a rapid and powerful general anesthetic.The clinical features are quick onset,short duration,rapid and stable wake up,less adverse reactions.The drug has been widely used in clinical subjects anesthesia and severe sedation.In recent years,it has been found that propofol has a strong inhibitory effect on the growth and invasion of HepG2 cell,but the specific molecular mechanism remains unclear.Epithelial-mesenchymal transition(EMT)refers to epithelial cells through a specific program into a biological process with interstitial phenotype cells,which confer the ability of cell metastasis and invasion,including stem cell characteristics,reducing apoptosis and promoting immunosuppression.At present,EMT has been considered to be a key factor in the recurrence and metastasis of liver cancer,and Wnt pathway has been an important inducer of EMT.Wnt signaling pathway can cause intracellular β-catenin accumulation.β-catenin is a multi-functional protein,which can interact with cadmium in the cell junction and be involved in the formation of adhesive tape.Free β-catenin can enter the nucleus,and then interact with TCF / LEF family transcription factor to regulate specific gene expression.Wnt signaling plays an important role in animal development.Usually,Wnt pathway activation is strictly monitored under normal physiological conditions.However,a large number of studies have shown that Wnt pathway is active in various tumors and is closely related to the development and progression of HCC.In recent years,it has been shown that propofol can decrease the mRNA expression of β-catenin in hepatocellular carcinoma to a certain extent.But whether it can really inhibit the Wnt signal pathway and produce anti-tumor effect is still not clear.In view of this,the aim of this study is to study the effect of propofol on the invasion and metastasis of human hepatocellular carcinoma cells and the effect of EMT on the Wnt / β-catenin signaling pathway,which is the new theoretical basis for the correct selection of intraoperative anesthetics.Methods:1.Hepatocellular carcinoma cells(HepG2、QGY-7703、SMMC-7721 、HCCLM3)were stimulated with propofol in gradually increasing(5 ug/ml,10 ug/ml,20 ug/ml),the effects of propofol on the migration and invasion of HCC cells were detected by Wound healing assay,Transwell matrix invasion assay and Three-dimensional spheroid invasion assay.2.Select a certain concentration(20ug/ml)of propofol to stimulate liver cancer cells HCCLM3,Western blot and immunofluorescence assay were used to detect the expression of EMT makers(E-cadherin and Vimentin).3.Select a certain concentration(20ug/ml)of propofol to stimulate liver cancer cells HCCLM3.The activity of Tcf-4 indirectly reflects the activation of β-catenin by luciferase reporter assay.Western blot was used to detect the the Subcellular localization of β-catenin.Real-time PCR was used to detect the mRNA expression levels of downstream genes(MYC,MMP9,TCF4)of Wnt/β-catenin signaling pathway.4.HCCLM3 cells were stimulated with propofol(20ug/ml)and 20 mmol/L Wnt pathway activator(Licl).Compared with the control group(propofol: 20ug/ml),the activation of β-catenin was detected by luciferase reporter assay.The migration and invasion of hepatocellular carcinoma cells were detected by Wound healing assay and Transwell invasion assay.Results:1.With the increase of propofol concentration,Wound healing assay showed that the migration ability of hepatocellular carcinoma cells decreased.Transwell invasion experiments and three-dimensional culture experiments showed that the invasive ability of hepatocellular carcinoma cells decreased.2.The expression of E-cadherin was up-regulated and the expression of Vimentin was down-regulated in HCCLM3 cells after propofol stimulation.3.The expression of β-catenin luciferase decreased in HCCLM3 cells,and the expression level of β-catenin protein in the nucleus was decreased,and the mRNA expression levels of MYC,MMP9 and TCF4 were decreased in the Wnt signaling pathway after propofol stimulation.4.The activity of β-catenin luciferase was enhanced by Wnt pathway activator(Licl)and propofol(20ug/ml),and the migration and invasion of HCC cells were enhanced.Conclusions:1.Propofol may inhibit the migration and invasion of HCC cells.2.Propofol may inhibit EMT in HCC cells.3.Propofol may suppress the migration and invasion of HCC cells through Wnt/β-catenin signaling pathway.