miR-642a-5p靶向KRT19调控甲状腺癌细胞TPC-1细胞周期的研究

Study on miR-642a-5p targeting KRT19 to regulate the cell cycle of thyroid cancer cell TPC-1

目的:探讨miRNA调控甲状腺癌细胞TPC-1细胞周期的潜在机制。方法:通过过表达甲状腺癌中异常表达的miRNA,检测甲状腺癌细胞TPC-1的细胞活力和细胞周期。通过miRDB在线分析miRNA的潜在靶标。通过siRNA敲低潜在靶标,检测甲状腺癌细胞TPC-1的细胞活力和细胞周期。通过荧光素酶报告实验检测miRNA与潜在靶标之间的关系。结果:过表达miR-642a-5p后,甲状腺癌细胞TPC-1的细胞活力显著下降(P<0.05)。敲低miR-642a-5p后,甲状腺癌细胞TPC-1的细胞活力显著上升(P<0.05)。过表达miR-642a-5p后,甲状腺癌细胞TPC-1处于G1期的细胞数量上升(P<0.05)、G2-M期和S期的细胞数量下降(P<0.05)。敲低miR-642a-5p后,甲状腺癌细胞TPC-1处于G1期的细胞数量下降(P<0.05)、G2-M期和S期的细胞数量上升(P<0.05)。当敲低KRT19后,甲状腺癌细胞TPC-1的细胞活力显著下降(P<0.05)。过表达miR-642a-5p后,KRT19的蛋白水平下降(P<0.05)。敲低miR-642a-5p后,KRT19的蛋白水平上升(P<0.05),miR-642a-5p靶向KRT19的3端非编码区(P<0.05)。敲低KRT19后,甲状腺癌细胞TPC-1的细胞活力下降(P<0.05)。过表达KRT19后,甲状腺癌细胞TPC-1的细胞活力上升(P<0.05)。敲低KRT19后,甲状腺癌细胞TPC-1处于G1期的细胞数量上升(P<0.05)、G2-M期和S期的细胞数量下降(P<0.05)。过表达KRT19后,甲状腺癌细胞TPC-1处于G1期的细胞数量下降(P<0.05)、G2-M期和S期的细胞数量上升(P<0.05)。同时敲低KRT19和miR-642a-5p后,甲状腺癌细胞TPC-1处于G1期的细胞数量、G2-M期和S期的细胞数量无显著变化(P>0.05)。同时过表达KRT19和miR-642a-5p后,甲状腺癌细胞TPC-1处于G1期的细胞数量、G2-M期和S期的细胞数量无显著变化(P>0.05)。结论:miR-642a-5p通过靶向KRT19 mRNA的3端非编码区,降低了KRT19的表达,调控甲状腺癌细胞TPC-1的细胞周期。

Objective:To explore the potential mechanism of miRNA regulating the cell cycle of thyroid cancer cell TPC-1.Methods:To detect the cell viability and cell cycle of TPC-1 thyroid cancer cells by overexpressing the abnormally expressed miRNA in thyroid cancer.Analyze potential targets of miRNA online through miRDB.Knock down potential targets by siRNA to detect the cell viability and cell cycle of thyroid cancer cells TPC-1.The relationship between miRNA and potential targets was detected by luciferase reporter experiment.Results:After overexpression of miR-642a-5p,the cell viability of thyroid cancer cell TPC-1 decreased significantly(P<0.05).After knocking down miR-642a-5p,the cell viability of thyroid cancer cells TPC-1 increased significantly(P<0.05).After overexpression of miR-642a-5p,the number of thyroid cancer TPC-1 cells in the G1 phase increased(P<0.05),and the number of cells in the G2-M and S phases decreased(P<0.05).After knocking down miR-642a-5p,the number of thyroid cancer cells in the G1 phase of TPC-1 decreased(P<0.05),and the number of cells in the G2-M and S phases increased(P<0.05).When KRT19 was knocked down,the cell viability of TPC-1 thyroid cancer cells decreased significantly(P<0.05).After overexpression of miR-642a-5p,the protein level of KRT19 decreased(P<0.05).After knocking down miR-642a-5p,the protein level of KRT19 increased(P<0.05).In addition,miR-642a-5p targets the 3 non-coding region of KRT19(P<0.05).After knocking down KRT19,the cell viability of thyroid cancer cells TPC-1 decreased(P<0.05).After overexpression of KRT19,the cell viability of thyroid cancer cells TPC-1 increased(P<0.05).After knocking down KRT19,the number of thyroid cancer TPC-1 cells in the G1 phase increased(P<0.05),and the number of cells in the G2-M and S phases decreased(P<0.05).After overexpression of KRT19,the number of TPC-1 cells in the G1 phase of thyroid cancer cells decreased(P<0.05),and the number of cells in the G2-M and S phases increased(P<0.05).After knocking down KRT19 and miR-642a-5p at the same time,the number of thyroid cancer TPC-1 cells in the G1 phase,the number of cells in the G2-M and S phases did not change significantly(P>0.05).After overexpression of KRT19 and miR-642a-5p at the same time,the number of thyroid cancer cells TPC-1 in the G1 phase,the number of cells in the G2-M and S phases did not change significantly(P>0.05).Conclusion:miR-642a-5p reduces the expression of KRT19 by targeting the 3 non-coding region of KRT19 mRNA,and subsequently regulates the cell cycle of thyroid cancer cells TPC-1.