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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 the migration and invasion of lung cancer by Notch1 signal pathway by miR-503. METHODS: The expression of BCYRN1 and miR-503 in different lung cancer cell lines was detected by q-PCR. The transfection efficiency of BCYRN1 + siRNA transfected with lentivirus BCYRN1 + siRNA was detected by immunofluorescence and q-PCR. The luciferase reporter gene detection of BCYRN1 and miR- 503 interaction; Transwell invasion assay and scratch assay were used to detect the invasion and migration of lung cancer cells after BCYRN1 silencing; Western blot was used to detect the expression of Notch1 signaling pathway after BCYRN1 silencing; Effect of tumorigenicity in nude mice. Results: The expression of BCYRN1 was the highest in lung cancer cell line H1299 and the expression level of miR-503 was relatively high. The expression of BCYRN1 + siRNA lentivirus could be effectively transfected into H1299 cells by immunofluorescence and mRNA levels. ’-UTR specific binding; silencing BCYRN1 can inhibit the invasion and migration of lung cancer H1299 cells; the expression of Notch1 pathway was down-regulated after BCYRN1 silencing; compared with NC group, the tumor volume and weight of BCYRN1-siRNA group mice Significantly reduced. CONCLUSION: BCYRN1 can regulate the invasion and migration of miR-503 through the Notch1 signal pathway in lung cancer H1299 cells.