The Effects Of Prunella Vulgaris On The Proliferation,Invasion And Migration Of BRAF Mutant Papillary Thyroid Carcinoma Cells By Regulating MicroRNA-146b-3p

Objective:In this study,B-CPAP cells with BRAF^V600Egene mutation were cultured in vitro;The expression of micro RNA-146b-3p in B-CPAP cells,the effect of Prunella vulgaris on the expression of micro RNA-146b-3p in B-CPAP cells and the changes of Prunella vulgaris on biological characteristics such as proliferation,invasion and migration of B-CPAP cells by affect the expression of micro RNA-146b-3p on B-CPAP cells were detected by a variety of biological molecular techniques.To search for a new target for the diagnosis and treatment of papillary thyroid cancer,and provide a theoretical basis for the clinical treatment of papillary thyroid cancer with Prunella vulgaris from the molecular level.Methods:Part one:The effect of Prunella vulgaris on micro RNA-146b-3p of BRAF^V600E gene mutant thyroid papillary cancer cells.1.Cell culture:B-CPAP cells of BRAF^V600E gene mutant thyroid papillary carcinoma cell line,Nthy-ori 3-1 cells of human thyroid normal cell line and EA cells of human umbilical vein endothelial cell line were cultured according to their corresponding suitable conditions.2.Detection of the drug safety of Prunella vulgaris:Different concentrations of Prunella vulgaris were applied to EA cells,and the safety of Prunella vulgaris was detected by CCK-8 experiment.3.The optimal concentration of cisplatin on B-CPAP cells was detected:Cisplatin solutions of different concentrations were treated on B-CPAP cells,and the optimal concentration of cisplatin on B-CPAP cells was detected by CCK-8 experiment.4.qRT-PCR was used to detect the expression of micro RNA-146b-3p in B-CPAP cells compared with Nthy-ori 3-1 cells,and the effect of Prunella vulgaris on the expression of micro RNA-146b-3p in B-CPAP cells.Part two:The effect of Prunella vulgaris on proliferation,invasion and migration of BRAF^V600E gene mutant papillary thyroid cancer cells by regulating the expression level of micro RNA-146b-3p.1.CCK-8 assay was used to detect the effects of Prunella vulgaris and silencing micro RNA-146b-3p on the proliferation of B-CPAP cells.2.Transwell assay was used to detect the effects of Prunella vulgaris and silencing micro RNA-146b-3p on the invasion of B-CPAP cells.3.Cell scratch assay was used to detect the effects of Prunella vulgare and silencing micro RNA-146b-3p on the migration of B-CPAP cells.Results:Part one:The effect of Prunella vulgaris on micro RNA-146b-3p of BRAF^V600E gene mutant thyroid papillary cancer cells.1.In this experiment,the concentration of 1.0 mg/m L Prunella vulgaris solution had no obvious toxic effect and was a safe dose.2.The results of CCK-8 assay indicated that the optimal concentration of cisplatin on B-CPAP cells was 10μg/m L（33.33μM）,which would be used for subsequent experiments.3.The qRT-PCR results showed that the expression level of micro RNA-146b-3p in B-CPAP cells was abnormal,which was about 4 times of that in normal thyroid cells.However,the traditional Chinese medicine Prunella vulgaris could significantly inhibit the expression of micro RNA-146b-3p in B-CPAP cells,with statistically significant differences in the results.Part two:Effect of Prunella vulgaris on proliferation,invasion and migration of BRAF^V600E gene mutant papillary thyroid cancer cells by regulating the expression level of micro RNA-146b-3p.1.Inhibition of micro RNA-146b-3p expression can inhibit the proliferation,invasion and migration ability of B-CPAP cells,especially the effect on invasion and migration.Therefore,micro RNA-146b-3p is expected to be a potential target for the diagnosis,treatment,and prognosis of BRAF^V600E gene mutant papillary thyroid carcinoma.2.Prunella vulgaris can significantly inhibit the proliferation ability of B-CPAP cells,but the inhibition effect was small by reducing the expression level of mi RNA-146b-3p.3.Prunella vulgaris can significantly reduce the invasiveness of B-CPAP cells by inhibiting the expression level of micro RNA-146b-3p.4.Prunella vulgaris can significantly reduce the migration of B-CPAP cells by inhibiting the expression level of micro RNA-146b-3p.Conclusion:The expression level of micro RNA-146b-3p in B-CPAP cells with BRAF^V600Egene mutation was abnormally higher than that in normal thyroid cells,and inhibition of the expression of micro RNA-146b-3p in B-CPAP cells could reduce the proliferation,invasion and migration ability of cancer cells.Therefore,micro RNA-146b-3p is expected to become a potential target for the diagnosis and treatment of papillary thyroid carcinoma.The traditional Chinese medicine prunella vulgaris can inhibit the proliferation,invasion and migration ability of B-CPAP of thyroid papillary cancer cells by reducing the expression level of micro RNA-146b-3p.