[摘要] 目的：探究长链非编码RNA（ long non-coding RNA，lncRNA)尿路上皮癌抗原1 ( urothelial carcinoma associated 1，UCA1) 调控非小细胞肺癌(NSCLC) A549 细胞增值、侵袭和迁移的作用及其机制。方法：NSCLC A549 细胞培养及慢病毒转染完成后，RT-PCR检测A549 细胞UCA1 表达水平，荧光素酶实验验证UCA1 和miR-185-5p 的靶向关系，MTT检测A549 细胞活性，Transwell 和划痕实验检测细胞侵袭和迁移能力；Western blotting 检测Wnt1/β-catenin 信号通路蛋白的表达。结果：sh-UCA1 能显著抑制UCA1 表达并促进miR-185-5p 表达(均P<0.05)，miR-185 inhibitor 可减弱sh-UCA1 对miR-185-5p 表达的促进作用(P<0.05)，UCA1 能明显抑制miR-185-5p 表达(P<0.05)，miR-185 mimic 可减弱UCA1 对miR-185-5p 表达的抑制作用(P<0.05)。荧光素酶报告实验表明UCA1 序列上有miR-185-5p 的结合位点。sh-UCA1 能显著抑制A549 细胞增殖、侵袭和迁移(均P<0.05)，并能降低信号通路蛋白Wnt1、β-catenin 信号和TCF-4 表达水平(均P<0.05)；miR-185 inhibitor 能显著减弱sh-UCA1 对A549 细胞的增殖、侵袭、迁移及对Wnt1/β-catenin 通路蛋白表达的抑制作用(P<0.05)。结论：UCA1 可通过靶向miR-185-5p 促进NSCLC细胞的增殖、侵袭和迁移，其作用机制与Wnt1/β-catenin信号通路激活有关。

[Abstract] Objective: To investigate the effect and underlying mechanism of Long non-coding RNA urothelial carcinoma associated 1 (lncRNA UCA1) on proliferation, invasion and migration of non-small cell lung cancer (NSCLC) A549 cells. Methods: NSCLS A549 cells were cultured and transfected with lentivirus; RT-PCR was employed to detect the levels of UCA1 in A549 cells. The relationship between UCA1 and miR-185-5p was validated by luciferase reporter assays. Cell viability of A549 cells was measured by MTT. Cell invasion and migration were determined by Transwell and Wound healing assay, respectively; and western blotting was performed for measuring the levels of Wnt1/β-catenin pathway-related proteins. Results: sh-UCA1 significantly decreased UCA1 expression and increased miR-185-5p expression in A549 cells (all P<0.05). miR-185 inhibitor attenuated the promotion effect of sh-UCA1 on miR-185-5p (P<0.05). UCA1 could significantly down-regulate miR-185-5p expression in A549 cells (P<0.05), which was reversed by miR-185 mimic (P<0.05). Luciferase reporter assay validated the binding site on UCA1 to link miR-185-5p. sh-UCA1 significantly inhibited cell proliferation, invasion and migration of A549 cells (all P<0.05), and also decreased the protein levels of Wnt1, β-catenin and TCF-4 notably (all P<0.05); however, miR-185 inhibitor attenuated such inhibitory effects of sh-UCA1 (P<0.05). Conclusion: UCA1 could promote proliferation, invasion and migration of A549 cells through targeting miR-185-5p, and the mechanisms might be related with activation of Wnt1/β-catenin pathway.

重庆市卫生局资助项目（No.2013-2-122）