| Thyroid cancer(TC)has become a common disease of the endocrine system and the incidence of this disease is increasing every year.TC can be classified into differentiated,undifferentiated and medullary carcinomas based on the type of pathology.The incidence of PTC is increasing year by year and the current treatment options are unavailable and risky,so further research into the pathogenesis of PTC and the search for new treatments is urgent.Autophagy is a phenomenon of "self-digestion" within cells.As one of the causes of cell death,autophagy is involved in the entire process of tumour development from onset to metastasis and plays a pivotal role,but whether autophagy plays a positive or negative role in cancer remains controversial.Although numerous studies have found that autophagy plays a dynamic role in inhibiting or promoting various aspects and processes in the development of different cancers,such as colorectal,liver,lung and gastric cancers,fewer studies have been reported on autophagy and thyroid cancer.Bufalin is one of the main active substances in traditional Chinese medicine and has been found to have antitumor activity in different types of cancer.The study of Bufalin and PTC has not been reported yet,therefore,in this experiment,Bufalin was used in papillary thyroid cancer BCPAP cells to investigate whether there is a correlation between the biological behavior of Bufalin and autophagy in PTC cells and to provide a new drug for the clinical management of PTC disease.Part I Effect of bufalin on the biological behaviour of BCPAP cells in papillary thyroid carcinomaObjective:In this study,we investigated the effect of Bufalin on the proliferation and migration of BCPAP in papillary thyroid carcinoma cells by using BCPAP cells as experimental subjects.Methods:1.CCK8 assay was performed to detect the effect of different concentrations of Bufalin on the proliferation ability of BCPAP cells after12 h,24h and 48 h.2.The cell scratch assay was performed to detect the changes of cell migration ability of BCPAP cells after 6h,12 h and 24 h of different concentrations of Bufalin.3.The experimental data were statistically analyzed by SPSS 26.0software,and all data were expressed as mean ± standard deviation(),and the difference was considered statistically significant at P < 0.05.Results:1.CCK8 assay results,Bufalin can inhibit the proliferation of BCPAP cells,and the difference was statistically significant(P < 0.05).2.The results of cell scratch assay,Bufalin effect on BCPAP cells after6 h,12h and 24 h,the cell migration ability decreased,and the scratch repair rate gradually decreased with increasing drug concentration,the difference was statistically significant(P < 0.05).Conclusions:1.Bufalin can inhibit the proliferation of BCPAP cells and induce apoptosis in cancer cells,which is significantly inhibited with the prolongation of drug action and the increase of drug concentration.2.Bufalin can inhibit the migration of BCPAP cells,which is significantly inhibited with the prolongation of drug action and the increase of drug concentration.Part II Effect of bufalin on autophagy in papillary thyroid cancer BCPAP cellsObjective:To further explore whether Bufalin inhibition of BCPAP cell proliferation and migration is related to autophagy based on the results of the first part of the experiment.Methods:1.The protein expression levels of autophagy-associated proteins Beclin-1 and P62 were detected by Western Blot after different concentrations of Bufalin had been applied to BCPAP cells for 24 h.2.The experimental data were statistically analyzed by SPSS 26.0software,and all data were expressed as mean ± standard deviation(),and the difference was considered statistically significant at P < 0.05.Results:Western Blot assay results,compared with the control group,the expression of protein Beclin-1 was significantly increased after Bufalin acted on BCPAP cells for 24h;the expression of protein P62 was significantly decreased,the difference was statistically significant(P < 0.05).Conclusions:1.Bufalin induced autophagy in papillary thyroid cancer cells BCPAP.2.Combined with the role of autophagy in cancer and the results of the present study,it was further hypothesized that Bufalin induced autophagy and apoptosis in BCPAP cells in coordination with each other,i.e.prompting autophagic apoptosis in cancer cells. |
