Objective:To investigate the effects and mechanisms of long non-coding RNA UCA1 on cell survival and migration of bladder cancer cell UM-UC-3 by targeting miR-582-5p.Methods:Cells were transferred with UCA1 shRNA(sh-UCA1)and(or)miR-582-5p,the transfection efficiency and level of miR-582-5p were detected by RT-PCR.The luciferase report assay was performed for validate the relationship of UCA1 and miR-582-5p.The cell viability was measured by CCK8 assay.Apoptosis was detected by flow cy-tometry.The metastatic ability was calculated by wound healing and Transwell assay.And the protein levels of proliferation-,apoptosis-and migration-related were determined by Western blot.Results:sh-UCA1 inhibited the expression of UCA1 and induced the expression of miR-582-5p(P<0.05),and miR-582-5p inhibitor alleviated the effect of UCA1 on miR-582-5p(P<0.05).The luciferase reporter assay indicated that there was miR-582-5p binding site on UCA1.Silencing of UCA1 inhibited proliferation of bladder cancer cells and the expression of Ki67,induced apoptosis and expression of cleaved caspase-3(P<0.05).Meanwhile,sh-UCA1 inhibited migration and invasion of bladder cancer cells coupled with decreasing VEGF(P<0.05).In addition,miR-582-5p inhibitor attenuated the effects of UCA1 on proliferation,apoptosis and migration(P<0.05).Conclusion:UCA1 promotes survival and migration of bladder cancer cells through targeting miR-582-5p.%目的:探究长链非编码RNA UCA1通过靶向miR-582-5p对膀胱癌细胞生存和运动能力的作用及作用机制.方法:用UCA1 shRNA(sh-UCA1)和(或)miR-582-5p inhibitor转染细胞,荧光定量检测转染效率及miR-582-5p的表达水平;荧光素酶报告实验确定UCA1和miR-582-5p的靶向关系;CCK8检测细胞活性,流式检测细胞凋亡情况,侵袭及划痕实验检测细胞侵袭迁移能力,免疫印迹检测细胞增殖、凋亡及迁移相关蛋白的表达.结果:sh-UCA1能显著降低膀胱癌细胞UCA1表达水平(P<0.05),促进miR-582-5p表达(P<0.05);miR-582-5p inhibitor能明显减弱sh-UCA1对miR-582-5p表达的促进作用(P<0.05);荧光素酶报告实验表明UCA1上有miR-582-5p的结合位点;沉默UCA1可显著抑制膀胱癌细胞增殖及Ki67的表达,促进细胞凋亡及cleaved caspase-3的表达(P<0.05);同时,sh-UCA1还能显著抑制膀胱癌细胞侵袭、迁移及VEGF的表达(P<0.05);此外,miR-582-5p inhibitor可显著减弱sh-UCA1对细胞增殖、凋亡及侵袭迁移能力的作用(P<0.05).结论:UCA1可通过靶向miR-582-5p增强膀胱癌UM-UC-3细胞的生存及运动能力.
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