Study On The Effect Of Artesunate Combined With Sorafenib On Cancer Cells

Objective:Sorafenib kinase inhibitor has been approved for the treatment of renal cell carcinoma,hepatocellular carcinoma(HCC)and flt3-itd-positive AML(acute myelogenous leukemia),which can significantly improve the overall survival rate of patients with advanced liver cancer.However,expensive long-term use can easily induce drug resistance,which limits the clinical use of SOR.Therefore,increasing the sensitivity of SOR is an important scientific issue in cancer treatment.It is known that sorafenib can significantly enhance the anti-cancer effect of sorafenib when combined with SOR and PI3K inhibitor.Moreover,it has been reported that artesunate(ARS)can play an anticancer role by acting on the PI3K/mTOR signaling pathway.The aim of this study was to elucidate the anti-cancer sensitization effect of artesunate on sorafenib and to explore its molecular mechanism.Methods:1.The OD values of SOR,ARS and SOR+ARS series concentrations were measured by MTS,and the proportion of SOR/ARS combined use was determined by IC50 values of each group.2.The number of viable and dead cells in CON,SOR,ARS and their combination groups were counted by trypan blue staining to further verify the inhibitory effect of combination of SOR and ARS on tumor proliferation.3.Scratch test was used to detect the effects of CON,SOR,ARS,SOR+ARS on the migration of SK-HEP1(human hepatic ascites adenocarcinoma cell)and HN30(human head and neck squamous cell carcinoma cell).4.Detection of apoptosis of SOR,ARS and combination drugs by flow cytometry after PI-Annxin V double staining5.The apoptotic effects of SOR,ARS and SOR+ARS on SK-HEP1 and HN30 cells were detected by PI-ANNXIN V double-staining flow cytometry.6.Western blot analysis was used to detect the protein expression levels of p-mTOR,PI3K and p-AKT in SOR,ARS and SOR+ARS groups.7.Data analysis was performed using Graphad 7.0 software.The IC50 calculation was performed using the Sigmoidal dose-response method.The measurement data between the two groups was compared using two-tailed Student's t test.Results:MTS experiments showed that the inhibition of proliferation of SK-HEP1 and HN30 cells in the combination of SOR and ARS was significantly higher than that in the single drug group with concentration-dependent effect.The apoptotic rates of SOR and ARS in SK-HEP1 and HN30 cells were higher than those in other groups(P<0.05);scratch test showed that the scratch healing rate of SOR and ARS combination group was slower than that of SOR and ARS stimulation group.The combination of SOR and ARS did not induce cell cycle.Western Blot results showed that artesunate and combination therapy could effectively down-regulate the expression of p-AKT and p-ERK in HN30 cells and Sk-hepl cells.Conclusion:In the cytological experiments of SK-HEP1 and HN30,ARS combined with SOR has positive synergistic effect,and can significantly enhance the inhibition of proliferation and migration of cancer cells,and promote the induction of apoptosis of SK-HEP1 and HN30 cells.ARS and SOR induce apoptosis of cancer cells by inhibiting PI3K/mTOR and MAPK/ERK signaling pathways.This study may provide a new way for cancer treatment.