Aqueous Extract Of Cordyceps Sinensis Potentiates The Antitumor Effect Of DDP And Attenuates Therapy-associated Toxicity And Studys On The Effect Of A375 Cells In Vitro

Cordyceps sinensis(Berk.)Sacc.is a famous medicinal fungus in Chinese herbal medicine.Because of its own characteristics and special growth environment,resulting in natural resources are scarce,disordered mining is exacerbated by the lack of resources,while the market demand is increasing,so to find a suitable substitute to become the focus of the study.After 10 years of research and development,Sunshine Lake Pharma Co.,LTD,has realized the artificial industrial breeding technology of Cordyceps sinensis.The culture of Cordyceps sinensis produced by this technology is not less than that of wild Cordyceps sinensis,indicating that Cordyceps sinensis has the potential to become a substitute for wild Cordyceps sinensis.We initially examined the in vitro antitumor effect of AECS,in vivo antitumor and toxicity-reduced effect of the combined treatment in Lewis xenografts mice model.Additionally,we evaluated the potential mechanisms of action of the combined treatment in Lewis xenografts mice model.Synergistic antitumor activity of the combined treatment was further evaluated in A549 xenografts zebrafish model,and toxicity-attenuated effect was also evaluated using zebrafish models.We found that AECS exhibited significant antitumor effect against various cancer cells in vitro.AECS,particularly AECS1,substantially potentiated the antitumor effect of DDP as evidenced by a significant decrease in tumor growth,a prolongation of the survival time,and also alleviated toxicity in Lewis xenografts mice model.The enhanced antitumor effect of the combined treatment in Lewis xenografts was accompanied by attenuated NF?B and AKT/mmp2/mmp9 pathways.Moreover,a markable increase of tumor inhibition in A549 xenografts zebrafish model was observed with the combined therapy,in comparison with DDP treatment alone,alleviated toxicity in zebrafish was also observed when treated with the combined therapy.Collectively,our results indicated that the combination of AECS1 and DDP should be a novel therapeutic modality to improve efficacy of DDP-based chemotherapy and reduce therapy-associated toxicity,possibly by inhibiting I?B?/NF?B and AKT/MMP2/MMP9 pathways.In order to study the antitumor mechanism of AECS in vitro,the experiment was carried out according to the following methods.First,we focused on the effect of AECS st on inhibiting the proliferation of A375 cells.MTT assay and clone formation assay were used to examine the inhibitory effect of AECS on A375 cells;Flow cytometry was used to detect its effect on cell cycle arrest.Second,we explored effect on the invasion and migration of A375 cell.Scratching test and transwell assay were used to detect its effect on migration and invasion;Cell adhesion assay was used to detect its effect on adhesion.Western blot was used to detect its effect on the expression of the proteins associated with cell cycle and MMPs.The results showed that AECS can significantly inhibit proliferation and colony formation of A375 cells in a dose-dependet manner and arrest cells at S phase.The scratch test and transwell showed that the artificial propagation of Cordyceps sinensis inhibited the migration and invasion of A375 cells.The adhesion test results showed that the adhesion ability of A3 7 5 cells was affected by the AECS.Western blot analysis showed that MMP-9,p-AKT,MMP-2,Bcl-2,CyclinE,CDK2,CDK4,protein expression decreased;Bax,P21 protein expression increased.The above results showed that AECS can significantly inhibit proliferation and invasion of A375 cell,and its mechanisms may arrest the cell cycle and activate of the AKT/MMP-2/MMP-9 pathway through regulating the expression of cell cycle associated proteins,MMPs family proteins,and p-AKT.