| Objective: By studying the effect of artesunate on thyroid cancer cells,establishing a transplanted tumor model in nude mice,discussing the anticancer effect of artesunate on thyroid cancer in vitro and in vivo,and exploring its mechanism,it is intended to provide a new reliable basis for predicting lymph node metastasis and prognosis evaluation of thyroid cancer patients,provide new ideas and perfect basis for tumor metastasis regulation mode,and provide a theoretical basis for the development of new treatment strategies.Methods: 1.In vitro experiments: The effects of ART on the viability,proliferation,migration,invasion,apoptosis and cell cycle of PTC cells were detected by CCK-8,Ed U,clone formation,scratch test,Transwell,flow cytometry and Western-blot.The PTC cells treated with ART were sequenced by whole genome second-generation sequencing technology,the enrichment pathway and target of ART were analyzed,and the expression of E2F1 related protein E2F1、c-Myc、Cylin E were detected.PTC cells overexpressed E2F1.The effects of ART on PTC cell viability,proliferation,invasion and apoptosis were detected.2.In vivo experiments: The nude mice model of PTC xenotransplantation was established and divided into control group and administration group.The mice in the administration group were given intragastric administration at the concentration of 5mg/kg.The mice in the control group were injected intraperitoneally with equal volume of normal saline.The tumor formation was observed after one week.After five weeks,the nude mice were killed by intraperitoneal injection of 100mg/kg sodium pentobarbital.The tumor xenografts were dissected,measured and weighed.Immunohistochemical staining was used to detect Ki67 and TUNEL in tumor tissues,extract protein and RNA from tumor tissues,detect the expression levels of apoptosis related proteins Bax,Bcl-2 and RAPR in tumor tissues,and detect the expression levels of E2F1 related proteins E2F1,c-myc and cyclin E in tumor tissues.Results: 1.In vitro experiments:(1)Artesunate treated PTC cells decreased cell viability,cell proliferation,migration and invasion,increased the expression of E-cadherin,and decreased the expression of N-cadherin and vimentin.The results of flow cytometry showed that the number of apoptosis increased,the expression of PARP and Bax protein increased in bht101 and tpc1 cells,and the expression of Bcl-2 protein decreased.Further statistical analysis showed that artesunate increased the expression of PARP and Bax protein in a dosedependent manner and inhibited the expression of Bcl-2 protein(P < 0.05).Artesunate blocked PTC cells in G0 / G1 phase.(2)By analyzing the whole genome,3052 significantly different genes were obtained,including 1086 up-regulated genes and 1966 down-regulated genes.124 enriched genes were obtained.ART inhibits E2F1 pathway in PTC cells,and E2F1 is the target gene of ART.(3)After art treatment of PTC cells,the expression levels of E2F1 and related proteins c-Myc and Cyclin E decreased.After overexpression of E2F1,the inhibitory effect of art on the activity,proliferation and invasion of PTC cells decreased significantly,and the promoting effect on the apoptosis of PTC cells decreased significantly.2.In vivo experiments:(1)The tumor volume and weight of nude mice in the administration group decreased.(2)The expression of Bax and RARP increased and the expression of Bcl-2 decreased in the tumor tissue of nude mice in the administration group.(3)Ki67 positive cells in tumor tissue decreased significantly and Tunel labeled apoptotic cells increased significantly in the administration group.The expressions of E2F1,c-Myc and Cyclin E in tumor tissues of mice in the administration group decreased.Conclusion: Artesunate inhibits the activity,proliferation,migration and invasion of PTC cells by combining with E2F1,promotes apoptosis,blocks PTC cells in G0 / G1 phase,inhibits tumor growth in xenograft nude mice,and induces tumor cell apoptosis in xenograft nude mice. |
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