长链非编码RNA UCA1 靶向miR-582-5p 对膀胱癌UM-UC-3细胞生存和运动能力的调节作用及机制研究

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。

Study on effects and mechanisms of long non-coding RNA UCA1 on cell survival and migration of bladder cell UM-UC-3 via targeting miR-582-5p

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 cytometry.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.