| Objective:Thyroid cancer is the most common in clinical endocrine system malignancies,and Papillary Thyroid Carcinoma（PTC）is the most common in all cases of thyroid cancer.Recent studies have shown that some mecha NISms of papillary thyroid carcinoma are related to changes in epigenetics.In addition,studies have shown that abnormal iodine nutrition affects thyroid papillary cancer cell lines more significantly than normal thyroid cells.Iodine nutrition levels,especially high iodine,also affect the genetic changes of thyroid cancer cells to a certain extent,but high iodine and The relationship between epigenetic changes in papillary thyroid carcinoma is still controversial.Therefore,whether high iodine can promote thyroid cancer by promoting epigenetic changes in thyroid needs to be explored.In this study,by analyzing the methylation and gene expression of TSHR and NIS promoters in papillary thyroid carcinoma cell lines,the relationship between the changes in the proliferation and motility of papillary thyroid carcinoma cells and high iodine was analyzed.Method:In this study,through dose screening,the common papillary cancer cell line TPC-1 was treated with different concentrations of potassium iodide（KI）for high iodine treatment,and the treatment dose was stronger;the methylation inhibitor decitabine（Decitabine,DAC）is formulated into the corresponding concentration solution,and the selected high iodine dose is used to treat the thyroid papillary carcinoma cell line TPC-1;after the cells are digested and resuspended,they are divided into 4 groups:PBS solvent control group,10-3 mmol/L KI treatment group,5μmol/L DAC control group,10-3 mmol/L KI+5μmol/L DAC intervention group.Place in a 37°C CO2 concentration 5%cell incubator for incubation.The CCK-8 test was used to test the cell proliferation ability,the scratch test evaluated the healing ability of the grouped cells,and the Transwell invasion test explored whether the cell invasion ability of different treatment groups was different;the methylation specific PCR（MSP）was used to detect the different treatment groups The methylation of the TSHR and NIS gene promoters in the medium;extracting total RNA from the cells,reverse transcription into c DNA,qualitatively exploring the expression of TSHR and NIS by gel electrophoresis,real-time fluorescence quantitative PCR（q RT-PCR）to detect TSHR and NIS m RNA Relative expression level.Results:（1）Screened by the CCK-8 cell proliferation experiment,the cell proliferation ability of the 10-3 mmol/L KI treatment dose group was enhanced compared with each exposure group,the difference was statistically significant（P<0.05）;and the methylation inhibitor intervention group and 10-3 mmol/L KI+5μmol/L DCA group motility and invasion ability of cells were reduced.Compared with the KI treatment group,the difference was statistically significant（P<0.05）.The cells in the 5μmol/L DCA group Compared with the solvent control group,there was no statistically significant difference in proliferation ability and exercise transfer ability.（2）In this experiment,both the TSHR and NIS genes in the solvent control group had DNA methylation in the promoter region,indicating that gene methylation was originally present in this papillary thyroid carcinoma cell line;from gel electrophoresis imaging,the TSHR gene The brightness of the amplified methylated band in the 10-3mmol/L KI+5μmol/L DAC treatment group was slightly lower than the brightness of the methylated band in the 10-3 mmol/L KI treatment group.Real-time fluorescence quantitative PCR showed that the solvent control The relative expression level of TSHR m RNA in the group was（1.91±0.31）,the relative expression level of TSHR m RNA in the 10-3 mmol/L KI dose group was（1.15±0.34）,and the relative expression level of TSHR m RNA in the 5μmol/L DCA treatment group was（2.01±0.15）,the relative expression level of TSHR m RNA in the 10-3 mmol/L KI+5μmol/L DCA treatment group was（1.80±0.54）,compared with the control group,the TSHR m RNA expression in the 10-3 mmol/L KI treatment group The level decreased and the difference was statistically significant（P<0.05）.The relative expression level of TSHR m RNA in the methylation inhibitor intervention group was increased.Compared with the KI treatment group,the difference was statistically significant（P<0.05）.（3）10-3 mmol/L KI+5μmol/L DAC treatment group,the brightness of the amplified methylation band was significantly lower than that of 10-3 mmol/L KI treatment Methylation band brightness,real-time fluorescence quantitative PCR showed that the relative expression level of NIS m RNA in the solvent control group was（3.86±0.67）,and the relative expression level of NIS m RNA in the 10-3 mmol/L KI treatment group was（2.28±1.01）,the relative expression level of NIS m RNA in the5μmol/L DCA intervention group was（6.95±3.14）,and the relative expression level of NIS m RNA in the 10-3 mmol/L KI+5μmol/L DCA intervention group was（9.81±4.62）.Compared with the treatment group,the relative expression level of NIS m RNA in the methylation inhibitor intervention group increased,and the difference was statistically significant（P<0.05）.Conclusion:On the basis of methylation of thyroid papillary cancer cell-related genes,high iodine can increase the degree of methylation of TSHR and NIS genes in the cells,reduce the relative expression level of thyroid cancer cell-related genes in thyroid papillary cancer cells,and promote Papillary thyroid cancer cell proliferation,motor metastasis and invasion ability;methylation inhibitor DAC can inhibit iodide to promote cancer growth,inhibit high iodine promote methylation,and partially restore TSHR and NIS genes in papillary thyroid cancer The expression in the cells can inhibit the proliferation and invasion ability of thyroid cancer cells,thereby slowing the pathogenesis of papillary thyroid cancer. |
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