【摘要】：目的:探討B(tài)CYRN1調(diào)控miR-503通過(guò)Notch1信號(hào)通路對(duì)肺癌遷移和侵襲的影響機(jī)制。方法:q PCR檢測(cè)不同肺癌細(xì)胞株中BCYRN1和miR-503的表達(dá)情況;免疫熒光和q PCR檢測(cè)慢病毒BCYRN1+siRNA轉(zhuǎn)染肺癌細(xì)胞的轉(zhuǎn)染效率;雙熒光素酶報(bào)告基因檢測(cè)BCYRN1與miR-503的相互作用;Transwell侵襲實(shí)驗(yàn)和劃痕實(shí)驗(yàn)檢測(cè)沉默BCYRN1后肺癌細(xì)胞侵襲和遷移能力的變化;Western blot檢測(cè)沉默BCYRN1后Notch1信號(hào)通路蛋白的表達(dá)情況;裸鼠皮下成瘤檢測(cè)沉默BCYRN1后肺癌細(xì)胞裸鼠體內(nèi)成瘤能力的影響。結(jié)果:在肺癌細(xì)胞H1299中BCYRN1表達(dá)水平最高,miR-503的表達(dá)水平相對(duì)較高;免疫熒光及mRNA水平證明BCYRN1+siRNA慢病毒可以有效轉(zhuǎn)染進(jìn)入H1299細(xì)胞內(nèi);BCYRN19能與miR-503的3'-UTR特異性結(jié)合;沉默BCYRN1可以抑制肺癌H1299細(xì)胞的侵襲和遷移能力;沉默BCYRN1后Notch1通路蛋白表達(dá)情況相應(yīng)下調(diào);與NC組相比,BCYRN1-siRNA組荷瘤小鼠腫瘤體積和重量都明顯減小。結(jié)論:BCYRN1可以靶向調(diào)節(jié)miR-503通過(guò)Notch1信號(hào)通路影響肺癌H1299細(xì)胞的侵襲和遷移能力。
[Abstract]:Objective: to investigate the mechanism of BCYRN1 regulating miR-503 on the migration and invasion of lung cancer through Notch1 signaling pathway. Methods: q PCR was used to detect the expression of BCYRN1 and miR-503 in different lung cancer cell lines, immunofluorescence and Q PCR were used to detect the transfection efficiency of lentivirus BCYRN1 siRNA transfected lung cancer cells, and double luciferase reporter gene was used to detect the interaction between BCYRN1 and miR-503. The changes of invasion and migration ability of lung cancer cells after BCYRN1 silencing were detected by Transwell invasion assay and scratch test. The expression of Notch1 signal pathway protein after BCYRN1 silencing was detected by; Western blot. The effect of BCYRN1 silencing on the tumorigenesis of lung cancer cells in nude mice was detected by subcutaneous tumorigenesis in nude mice. Results: the expression of BCYRN1 in H1299 cells was the highest and the expression level of miR-503 was relatively high. Immunofluorescence and mRNA levels showed that BCYRN1 siRNA lentivirus could be transfected into H1299 cells effectively, and BCYRN19 could specifically bind to 3'-UTR of miR-503. Silencing BCYRN1 inhibited 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, the tumor volume and weight of BCYRN1-siRNA group decreased significantly. Conclusion: BCYRN1 can target regulate the invasion and migration of lung cancer H1299 cells by miR-503 through Notch1 signaling pathway.
【作者單位】： 河南省新鄉(xiāng)醫(yī)學(xué)院醫(yī)學(xué)檢驗(yàn)學(xué)院和分子診斷與醫(yī)學(xué)檢驗(yàn)技術(shù)河南省協(xié)同創(chuàng)新中心;河南省新鄉(xiāng)醫(yī)學(xué)院第一附屬醫(yī)院腫瘤科;
【基金】：國(guó)家自然科學(xué)基金(No.31301135) 河南省自然科學(xué)基金(No.162300410211) 河南省科技攻關(guān)計(jì)劃項(xiàng)目(No.201203068)
【分類號(hào)】：R734.2

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