Effect Of 5-Aza-CdR On Biological Behavior Of Human Papillary Thyroid Carcinoma Cell Line

Objective Methylation of tumor suppressor gene leads to gene silencing play an important role in the occurrence and development of thyroid cancer,and demethylation preparation can reproduce the expression of silencing tumor suppressor gene and continue to exert its anti-tumor biological effect.This study aimed to explore the effects of 5-Aza-CdR on the biological behavior of human papillary thyroid carcinoma cells from a molecular perspective,and provide new clues and ways for molecular therapy of thyroid cancer.Methods Different concentrations of 5-Aza-CdR were applied to papillary thyroid carcinoma TPC-1 cells and normal thyroid Nthy-ori-3 cells.Cell proliferation was detected by MTT colorimetric assay,and cell cycle was analyzed by flow cytometry.The death and scratch healing experiments were performed to analyze the cell migration ability.The PTEN gene protein was detected by Western blot,and the changes of cell function before and after treatment were observed.Results(1)Inhibition of proliferation of human papillary thyroid carcinoma TPC-1 cells at different concentrations(1μmol/L,5μmol/L,10μmol/L)of 5-Aza-CdR at 24 h,48h and 72 h compared with before administration.The rate was higher,the difference was significant(P<0.001),and with the prolongation of the action time(24h,48 h,72h)and the increase of concentration,the inhibition of cell proliferation was enhanced,while the human normal thyroid Nthy-ori-3 There was no significant change in the cell line;(2)Compared with before administration,different concentrations of 5-Aza-CdR acted on human papillary thyroid carcinoma TPC-1 cells for 24 h and 48 h,and the proportion of cells in G1 phase(DNA content)increased gradually.The proportion of cells in S phase decreased gradually,the difference was statistically significant(P<0.01),with significant time and concentration dependence;while Nthy-ori-3 cell line showed no significant change;(3)different from before use The concentration of 5-Aza-CdR was applied to human papillary thyroid carcinoma TPC-1 cells for 24 h.After 48 h,the apoptotic rate of cells also increased significantly,showing a concentration-dependent effect,and the difference was statistically significant(P<0.05).There was no significant change in the Nthy-ori-3 cell line;(4)the concentration of 5-Aza-CdR was comparable to that before administration.The migration of human papillary thyroid carcinoma TPC-1 cells,and the stronger the inhibition ability with increasing concentration,the difference was statistically significant(P<0.05);(5)10μmol/L 5-Aza-CdR compared with before administration.After treatment of human papillary thyroid carcinoma cell line TPC-1 for24 h,the expression level of PTEN gene protein increased,the difference was statistically significant(P<0.001).In human normal thyroid Nthy-ori-3 cells,PTEN expression was very little,and There was no change before and after dosing(P>0.05).Conclusion5-Aza-CdR can significantly inhibit the proliferation and metastasis of human papillary thyroid carcinoma TPC-1 cells,arrest cell cycle,induce apoptosis,and exert its anti-tumor effect,and with 5-Aza-CdR The increase of concentration and the prolongation of action time increased,and 5-Aza-CdR induced the restoration of protein expression in the silenced PTEN gene and exert the effect of inhibiting tumor cell growth.