摘要
目的:探究长链非编码RNA(long non-coding RNA,lnc RNA)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mi R-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mi R-185-5p表达(均P<0.05),mi R-185 inhibitor可减弱sh-UCA1对mi R-185-5p表达的促进作用(P<0.05),UCA1能明显抑制mi R-185-5p表达(P<0.05),mi R-185 mimic可减弱UCA1对mi R-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mi R-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);mi R-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mi R-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。
Objective： To investigate the effect and underlying mechanism of Long non-coding RNA urothelial carcinoma associated 1（lnc RNA 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 mi R-185-5 p 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 mi R-185-5 p expression in A549 cells（all P〈0.05）. mi R-185 inhibitor attenuated the promotion effect of sh-UCA1 on mi R-185-5 p（P〈0.05）. UCA1 could significantly down-regulate mi R-185-5 p expression in A549 cells（P〈0.05）, which was reversed by mi R-185 mimic（P〈0.05）. Luciferase reporter assay validated the binding site on UCA1 to link mi R-185-5 p. 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, mi R-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 mi R-185-5 p, and the mechanisms might be related with activation of Wnt1/β-catenin pathway.
作者
蔡华荣
王志强
江跃全
CAI Huarong;WANG Zhiqiang;JIANG Yuequan(Department of Thoracic Surgery, Affiliated Cancer Hospital of Chongqing Universit;Chongqing Cancer Institute, Chongqing 400030, Chin)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2018年第6期555-561,共7页
Chinese Journal of Cancer Biotherapy
基金
重庆市卫生局资助项目(No.2013-2-122)~~
