Research Of Anti-Esophageal Cancer Effects And Mechanisms Of Dehydrocostus Lactone

Cancer is a serious threat to human health,and it is still a top priority to find effective cancer treatment methods.One of the main methods of treating cancer is chemotherapy,but the current chemotherapeutic drugs have strong toxic and side effects,and easily make patients resistant to drug resistance,which greatly reduces the efficacy of chemotherapeutic drugs.The natural products extracted and isolated from medicinal plants have attracted the attention of researchers due to their advantages such as easy availability and mild side effects,and the discovery of new anticancer drugs from medicinal plants has become a current research hotspot.Esophageal cancer is one of the most fatal malignancies worldwide.DHL derived from the dried roots of Saussurea costus(Falc.)Lipech is a sesquiterpene lactone compound that exerts anticancer activities.However,only one report on the anti-esophageal cancer activity of DHL can be retrieved currently,and the deeper mechanisms need further research.In this study,DHL was obtained to evaluate its anti-esophageal cancer ability and underlying mechanism in vitro and in vivo,which will provide a more basis for the drug treatment of esophageal cancer.The dried roots of Saussurea costus(Falc.)Lipech were crushed and extracted by percolation extraction,and then the monomer compound DHL was separated and purified by macroporous resin column chromatography and other methods.In vitro,MTT assay was used for the detection of cell viability,the wound healing assay was used to detect cell migration ability,Hoechst33258 staining and Annexin Ⅴ-FITC/PI double staining(flow cytometry assay)were used to detect cell apoptosis,ROS detection kit and JC-1 kit were used to detect the level of intracellular reactive oxygen species and mitochondrial membrane potential,respectively,and Western blot assay was used to detect related pathway proteins to determine the specific mechanism of DHL against esophageal cancer.In vivo,BALB/C nude mice were injected subcutaneously with Eca109 cells to establish a Eca109 tumor-bearing nude mouse model to further verify the anti-tumor activity and mechanism of DHL in vivo.HE staining,TUNEL and IHC were used to explore the effect of DHL on tumor tissues.Similarly,Western blot was used to to further identify specific signaling pathways and targets of DHL acting on esophageal cancer cells in vivo.Our research showed that DHL inhibited the proliferation and migration of Eca109 and KYSE150 esophageal cancer cells in a time-and dose-dependent manner in vitro,and it showed less cytotoxic to human normal embryonic kidney HEK293 cells.In vivo,DHL inhibited the growth of Eca109 tumor xenografts in a dose-dependent manner with no significant signs of toxicity in the organs of nude mice.Mechanistically,DHL induced esophageal cancer cells apoptosis in vivo and in vitro through the mitochondrial pathway,and the PI3K/Akt/Bad pathway participated in this process.DHL induced autophagy in esophageal cancer cells by up-regulating the expression of LC3-Ⅱ and down-regulating the expression of p62 in vivo and in vitro.In addition,treatment with DHL could significantly activate ROS in cells,leading to mitochondrial damage.The ROS scavenger NAC inhibited DHL-induced apoptosis and autophagy,the pancaspase inhibitor Z-VAD-FMK diminished DHL-induced autophagy,but the autophagy inhibitor 3-MA had no effect on DHL-induced apoptosis.The above results indicated that DHL inhibited esophageal cancer cells through ROS-mediated apoptosis and autophagy in vivo and in vitro,and it can be considered a potential chemotherapeutic agent for esophageal cancer.