The Influence Of Parthenolide On Apoptosis And Autophagy Of Human Thyroid Carcinoma Cell

Objective:Thyroid cancer is the most common endocrine malignant tumor.Compared with other tumors,the prognosis of thyroid cancer is relatively good,but its growth rate is gradually increasing in recent years.At present,the treatment of thyroid cancer mainly includes surgical resection,radioactive iodine ablation and drug treatment.Many indicators affect its therapeutic effects,such as age,gender,tumor size and extra thyroid expansion.With the differentiation of thyroid cancer,traditional surgery and radioactive iodine ablation are no longer effective for partially differentiated thyroid cancer.Studying new drugs that can alleviate thyroid cancer is of great significance for the treatment of thyroid cancer.Between 1981 and 2014,49%of the anti-tumor drugs approved for marketing worldwide were directly or indirectly extracted from natural products,especially botanicals.The parthenolide is first extracted from the herbaceous plant Tanacetum parthenium.It is a sesquiterpene lactone with anti-inflammatory,immune-modulating,anti-asthmatic and anti-tumor effects.Currently,the anti-tumor effect of parthenolide on thyroid cancer has not been fully elucidated.In this study,the effects of parthenolide on apoptosis and autophagy of thyroid carcinoma BCPAP cells were studied by in vitro cell experiments,in order to provide reference for clinical treatmentMethods:In this experiment,the PTL was first divided into different concentrations(0,4,6,8,10,12 ?mol/L)to treat BCPAP for 24 h and 48 h,respectively.The Half-inhibitory concentration and survival rate were calculated by MTS method at different times.PTL was divided into(0,8,10,12 ?mol/L)to treat BCPAP cells for 24 h,apoptosis was detected by Hoechst 33258 fluorescent staining and Annexin V-FITC/PI double staining flow cytometry;The mitochondrial membrane potential were detected by JC-1 fluorescent probe flow cytometry;The intracellular reactive oxygen species were detected by DCFH-DA fluorescent probe flow cytometry;The intracellular autophagy marker LC3I/II were detected by immunofluorescence staining;The expression of autophagy-associated protein,LC3II and Beclinl,were detected by Western Blot;The effect of PTL on apoptosis of thyroid carcinoma BCPAP cells after adding autophagy inhibitor by Annexin V-FITC/PI double staining flow cytometry Results:With the increase of PTL concentration,the activity of BCPAP decreased,there were statistical differences between the groups(**P<0.01,***P<0.0001).At the same concentration,the cell viability in the 48h group was significantly lower than that in the 24 h group,indicating that the inhibitory effect of PTL on BCPAP cell proliferation was in a time-and dose-dependent manner.With the increase of the dose of the drug,the number of cells gradually decreased,the cell morphology gradually became irregular,the fluorescence gradually became dense,and the normal nuclear morphology was lost,the apoptosis rate gradually increased.The 12 ?mol/L group was significantly higher than the other groups.The difference between the groups was statistically significant(*P<0.05,***P<0.0001).With the increase of the concentration of the drug,the green fluorescence of the cells gradually increased,and the ratio of the red fluorescence/green fluorescence of the cells gradually decreased,the mitochondrial membrane potential dropped,and the amount of fluorescence in the R2 region gradually increased,the release of reactive oxygen species increased,and the difference between the groups was statistically significant(P<0.0001).Compared with the control group,the cell number decreased gradually with the increase of the concentration of the drug,and the fluorescence expression of LC3 I/II increased gradually,the labeling proteins LC3II and Beclinl of autophagy gradually increased with the increase of the concentration of parthenolide in the medium.The difference was statistically significant(P<0.05).After using the autophagy inhibitor 3-MA,flow cytometry showed that there was no significant difference between the 3-MA and control groups(P>0.05).The apoptotic rate of the 3-MA+PTL combination group was lower than that of the PTL group alone,and there was significant difference between the groups(P<0.05).This indicates that inhibition of autophagy levels in BCPAP cells induced by PTL can inhibit cell apoptosisConclusion:1.PTL can significantly inhibit the proliferation of thyroid cancer BCPAP cells in a time-and dose-dependent manner2.PTL can promote the apoptosis of thyroid cancer BCPAP cells and increase the autophagy level of cells3.High levels of autophagy are important conditions for PTL to induce apoptosis in thyroid cancer BCPAP cells4.ROS may be an important factor in PTL-induced apoptosis and autophagy in thyroid cancer BCPAP cells.