BCYRN1调控miR-503通过Notch1信号通路对肺癌迁移和侵袭的影响

Effects of BCYRN1 on miR-503 on migration and invasion of lung cancer by Notch1 pathway

目的:探讨BCYRN1调控miR-503通过Notch1信号通路对肺癌迁移和侵袭的影响机制。方法:q PCR检测不同肺癌细胞株中BCYRN1和miR-503的表达情况;免疫荧光和q PCR检测慢病毒BCYRN1+siRNA转染肺癌细胞的转染效率;双荧光素酶报告基因检测BCYRN1与miR-503的相互作用;Transwell侵袭实验和划痕实验检测沉默BCYRN1后肺癌细胞侵袭和迁移能力的变化;Western blot检测沉默BCYRN1后Notch1信号通路蛋白的表达情况;裸鼠皮下成瘤检测沉默BCYRN1后肺癌细胞裸鼠体内成瘤能力的影响。结果:在肺癌细胞H1299中BCYRN1表达水平最高,miR-503的表达水平相对较高;免疫荧光及mRNA水平证明BCYRN1+siRNA慢病毒可以有效转染进入H1299细胞内;BCYRN19能与miR-503的3'-UTR特异性结合;沉默BCYRN1可以抑制肺癌H1299细胞的侵袭和迁移能力;沉默BCYRN1后Notch1通路蛋白表达情况相应下调;与NC组相比,BCYRN1-siRNA组荷瘤小鼠肿瘤体积和重量都明显减小。结论:BCYRN1可以靶向调节miR-503通过Notch1信号通路影响肺癌H1299细胞的侵袭和迁移能力。

Objective： To investigate the mechanism of BCYRN1 regulating miR-503 through Notch1 signaling pathway in the migration and invasion of lung cancer. Methods： The expression of BCYRN1 and miR-503 in different lung cancer cell lines were detected by q PCR. Immunofluorescence and q PCR were used to detect the transfection efficiency of lentivirus BCYRN1 ＋ siRNA transfected lung cancer cells. Double luciferase reporter gene was used to detect the interaction between BCYRN1 and miR-503. Transwell invasion test and scratch test were used to detect the invasion and migration of lung cancer cells after silencing BCYRN1. Western blot was used to detect the expression of Notch1 signaling pathway silence BCYRN1. The effect of silencing BCYRN1 on lung cancer cells in nude mice was measured by subcutaneous tumor formation in nude mice. Results： The expression level of BCYRN1 was the highest in lung cancer cell H1299,and the expression level of miR-503 was relatively high. Immunofluorescence and mRNA levels demonstrated that BCYRN1 ＋ siRNA lentivirus could be effectively transfected into H1299 cells; BCYRN19 binds specifically to the 3＇-UTR of miR-503; silencing BCYRN1 could inhibit the invasion and migration of lung cancer H1299 cells; the expression of Notch1 pathway protein was down-regulated after silencing BCYRN1. Compared with NC group,tumor volume and weight of BCYRN1-siRNA group were significantly decreased. Conclusion： BCYRN1 can target the regulation of miR-503 through Notch1 signaling pathway in the invasion and migration of lung cancer H1299 cells.