The Inhibition Effect Of Propofol On Hepatocellular Carcinoma Growth And Metastasis:Mechanism Study

Part â…  The study of propofol inhibition the growth and metastasis of hepatocellular carcinoma cells In vitroBackground:Propofol is one of the most commonly intravenous anesthetics used in the tumor resection operation. The inhibitory effect of propofol on the growth and metastasis of tumor cells and its effect on tumor immunity get gradually noticed. It was first reported that propofol can inhibit the invasion ability of human cervical cancer, fibrosarcoma, osteosarcoma cell line in vitro. Then, the experiments in vivo and vitro show that propofol can inhibit breast cancer cell growth, and, studies in vitro about esophageal cancer and lung cance have also confirmed the antitumor effect of propofol. However, the effect of propofol on hepatocellular carcinomar cells has not been reported. Objective:The purpose of this study is demonstrate the effect of propofol on hepatocellular carcinoma cell growth and invasion.Methods:1. Different concentrations of propofol and hepatocellular carcinoma HepG2cells normal liver cell line L-02were co-cultivated.2. Cell viability was measured by MTT.3. Enzyme linked immunosorbent assay was used to detect the expression of Caspase8, Caspase9.4. Transwell cell by cell invasion assay.5. Matrix metalloprotease-9was measured by gelatin zymography.6. Western blot the determination of the expression of MMP-9protein.Results:1. Propofol inhibited the growth of hepatocellular carcinoma cell line HepG2and normal liver cell line L02. Among them, the normal cells showed stronger resistance to propofol.2. Propofol induced HepG2apoptosis, accompanied by increased Caspase-8and Caspase-9activity. Moreover, the apoptosis rate of HepG2was positively correlated with propofol concentration.3.5ug/ml and lOug/ml of propofol could significantly decrease HepG2cell adhesion.The expression level of MMP-9was dose-dependently decreased in HepG2cells after treated with5,10g/ml propofol for16hours. Conclusions:Clinically relevant concentrations of propofol can dose-dependently induce the apoptosis of HepG2cells, and increase the activation of apoptosis protein Caspase-8and Caspase-9. Propofol can inhibit the invasion ability of HepG2cells, the activity of MMP-9in HepG2cells was also obviously inhibited. Effect of propofol on tumor cells are diversity, so the details of the mechanism needs further study. Part II Propofol exerts anti-hepatocellular carcinoma by up-graduating miR-142-3p from macrophage and microvesicleBackground:The first part of this study found that propofol can inhibit the growth and metastasis of hepatocellular carcinoma in vitro, but it is not enough to guide the clinical practice, further animal experiment study of propofol need to be done, especially the effect of propofol on immune function.Macrophage has heterogeneity and diversity. In different microenvironment stimuli, it can differentiate into the opposite biological function subset. Tumor associated macrophages, which is the most abundant immune cells in the tumor microenvironment, raised by tumor cells and it is formed the tumor immune microenvironment with other stromal cells together. The higher levels of macrophage colony stimulating factor in hepatocellular carcinoma patients hinted recurrence after resection of hepatocellular carcinoma. Research on a variety of tumor showed that miRNA plays an important role in the pathogenesis of tumors, miR-142-3p plays an important role in the inhibition of cell migration and invasion of hepatocellular carcinoma, and enhance the expression of miR-142-3p can inhibit the differentiation of tumor associated macrophages.Microvesicles loaded many bioactive molecules including miRNA. Microvesicles released by induced macrophages, and through the microvesicles, information material was transfered to the surrounding tumor cells.Studies have demonstrated that propofol can show the anti-glioma effect by affecting macrophages to release reactive oxygen species familyt, this part of the study supposed that propofol activate the tumor associated macrophages, up-regulate miR-142-3p and stimulate tumor associated macrophages to secret microvesicles full of miR-142-3p.Furthermore, propofol exerts anti-tumor effect by microbubbles.Objective:The purpose of this study is demonstrate the effect of propofol exerting anti-hepatocellular carcinoma by microvesicle-mediated transfer of miR-142-3p from macrophage to cancer cellsMethods:1. Establishment of Hepal-6mouse subcutaneous transplantation tumor model; Differential centrifugal separation of plasma MVs. According to the different treatment groups, tumor bearing mice were divided into8groups:(1)20mg/kg propofol;(2) l00g/kg clodrolip; (3)20mg/kg propofol+100g/kg clodrolip;(4)50mg/kg propofol;(5)50mg/kg propofol+100g/kg clodrolip;(6)20g MVs;(7) miR-142-3p inhibitor;(8)20mg/kg propofol+miR-142-3p inhibitors. The tumor volume and weight were measured in tumor bearing mice.2. Isolation of mouse TAMs and co-cultured with Hepal-6cells, Transwell cell tested cell invasion assay.3. The level of miR-142-3p in macrophages, micro vesicle and Hepal-6cells were measured by Real time fluorescence quantitative PCR. The level of miR-142-3p of ipofectamine TM2000transfection miR-142-3p inhibitor were also measured.Results:1. Propofol inhibited HCC growth in vivo.2. Co-cultivation with TAMs from propofol-treated HCC tissue increased miR-142-3p levels in HCC cells.3. Propofol stimulated miR-142-3p shuttling from macrophages to HCC cells.4. MVs collected from the plasma of tumor-bearing mice injected with propofol enhance anti-HCC in vivo.5. miR-142-3p was involved in the effect of propofol on anti-tumor activity.Conclusions:We found that propofol could inhibit growth and metastasis of hepatocellular carcinoma cells by up-regulating the miR-142-3p expression in tumor associated macrophages and secreted microvesicle.