The Impact Of Caudatin On Biological Behavior In Seminoma Cell Line

Testicular Germ Cell Tumor(TGCT)is a kind of tumor that attends to occur in young men,and the incidence has been increased year by year.Traditional therapeutic methods combined surgery with chemotherapy and radiotherapy,but chemotherapy and radiotherapy bring great side effects.In recent years,many studies have shown that curcumin,paclitaxel,and many other ingredients in traditional Chinese medicine can inhibit the growth of a variety of tumor cells,and these drugs are famous for their low toxicity,so they are probably to be widely used.According to previous studies,a kind of C21 steroidal glycosides named Caudatin extracted from Cynanchum caudatum Max has been considered to inhibit gastric cancer cell proliferation,promote apoptosis.But whether caudatin plays the same role in testicular germ cell tumor was not reported.TGCT can be classified as seminoma and non-seminoma.In our study,seminoma cell lines NTera-2 and GC-1 were treated with different doses of caudatin,both MTT assay and colony formation assay demonstrated that caudatin could inhibit their proliferation in a dose-dependent manner.Then we observed apoptosis in NTera-2 cells at cellular and molecular levels when treated with different doses of caudatin.By western blotting and staining with hoechst33258,we demonstrated that caudatin induced cell apoptosis.By transwell assay and wound-healing assay,caudatin also inhibited migration of NTera-2 cells in a dose-dependent manner.Besides,TGCT marker gene AP-2y and OCT4 were down-regulated in caudatin treated NTera-2 cells.To investigate the mechanism further,we conducted the wound-healing assay in NTera-2 cells after transfected with si-AP-2y which can decrease the expression of AP-2?.Results showed that the ability of cell migration was decreased as well.Also,we conducted the sub-acute toxicity experiments to investigate the toxicity of caudatin in Kunming mice.The mice were treated with certain doses of caudatin through intraperitoneal injection for 14 consecutive days.Then blood were taken from eyeball and hematology and serum biochemical indexes were evaluated.The results showed there was no significant difference between different doses and proved that caudatin had low toxicity.In conclusion,our study showed that caudatin was a kind of antitumor drug with low toxicity which could inhibit cell proliferation,promote apoptosis and inhibit cell migration by suppressing the expression of AP-2y in seminoma cell line NTera-2,and these can provide theoretical basis for clinical treatment of testicular germ cell tumor.