LncRNA UCA1靶向miR-520a-5p对膀胱癌细胞增殖、凋亡及放射敏感性的影响OA
Effects of LncRNA UCA1 on proliferation,apoptosis and radiosensitivity of bladder cancer cells by targeting miR-520a-5p
目的 探讨长链非编码核糖核酸尿路上皮癌胚抗原1(LncRNA UCA1)调节miR-520a-5p对膀胱癌细胞增殖、凋亡及放疗敏感性的影响.方法 RT-qPCR检测正常膀胱上皮细胞株(HCV-29)和人膀胱癌细胞株(SW780、HT1376、BIU87 和 T24)LncRNA UCA1、miR-520a-5p 表达,使用不同放疗剂量(0～8 Gy)处理T24细胞,检测细胞增殖活性,将T24细胞分为对照(Control)组、sh-NC组、sh-UCA1组、sh-UCA1+anti-miR-NC组和sh-UCA1+anti-miR-520a-5p组,各组细胞使用放疗剂量2 Gy处理,分别检测各组细胞增殖、凋亡及细胞周期蛋白D1(CyclinD 1)、Ki-67抗原(Ki-67)、Bcl-2相关X蛋白(Bax)、胱天蛋白酶-3(Caspase-3)蛋白表达;双荧光素酶报告基因实验、原位杂交(FISH)实验、RNA免疫共沉淀实验验证LncRNA UCA1与miR-520a-5p的靶向关系.建立裸鼠肿瘤异种移植模型,验证LncRNA UCA1对膀胱癌放疗敏感性的影响.结果 在膀胱癌细胞系中LncRNA UCA1表达升高,miR-520a-5p表达降低(P＜0.05);不同放疗剂量(0～8 Gy)处理T24细胞后,随着放射剂量增加,sh-UCA1组细胞增殖活性明显低于sh-NC组(P＜0.05);选择2 Gy放疗剂量处理T24细胞.在T24细胞中,敲低LncRNA UCA1可显著抑制细胞增殖,促进细胞凋亡并增加放疗敏感性,下调CyclinD 1、Ki-67 表达,上调 Bax、Caspase-3 表达(P＜0.05);miR-520a-5p 拮抗剂可减弱敲低 LncRNA UCA1 对膀胱癌细胞增殖、凋亡及放疗敏感性的影响(P＜0.05);LncRNA UCA1可靶向负调控miR-520a-5p表达.敲低LncRNA UCA1联合放疗可显著抑制膀胱癌移植瘤生长,增加放疗敏感性(P＜0.05).结论 LncRNA UCA1在膀胱癌细胞中表达上调,敲低LncRNA UCA1可通过上调miR-520a-5p表达,抑制细胞增殖并诱导细胞凋亡,增强膀胱癌细胞放疗敏感性.
Objective To investigate the effects of urothelial carcinoma antigen 1 long non-coding RNA(LncRNA UCA1)on the proliferation,apoptosis,and radiosensitivity of bladder cancer cells by regulating miR-520a-5p.Methods LncRNA UCA1 and miR-520a-5p were detected in normal bladder epithelial(HCV-29)and human bladder cancer(SW780,HT1376,BIU87 and T24)cell lines by quantitative reverse transcription-polymerase chain reaction.T24 cells were treated with different radiation doses(0～8 Gy)and cell proliferation activity was detected.T24 cells were assigned to Control,sh-NC,sh-UCA1,sh-UCA1+anti-miR-NC,and sh-UCA1+anti-miR-520a-5p groups.Cells in each group were treated with a radiation dose of 2 Gy,and cell proliferation,apoptosis,and cyclin D1,Ki-67,Bax,and Caspase-3 protein expression levels were detected.The targeting relationship between LncRNA UCA1 and miR-520a-5p was verified by dual-luciferase reporter gene,fluorescence in situ hybridization,and RNA immunoprecipitation assays.A nude mouse tumor xenotransplantation model was established,and the effect of LncRNA UCA1 on the radiosensitivity of bladder cancer was verified.Results LncRNA UCA1 was increased and miR-520a-5p was decreased in bladder cancer cell lines.Treating T24 cells with different radiation doses(0～8 Gy)reduced cell proliferation activity in the sh-UCA1 group compared with the sh-NC group,in line with increasing radiation dose(P＜0.05).T24 cells and a radiation dose of 2 Gy were therefore selected for subsequent experiments.Knockdown of LncRNA UCA1 in T24 cells inhibited cell proliferation,promoted cell apoptosis,increased radiosensitivity,down-regulated cyclin D1 and Ki-67,and upregulated Bax and Caspase-3(P＜0.05).miR-520a-5p antagonist weakened the impacts of LncRNA UCA1 knockdown on the proliferation,apoptosis,and radiosensitivity of bladder cancer cells(P＜0.05).LncRNA UCA1 targeted the negative regulation of miR-520a-5p.Knockdown of LncRNA UCA1 combined with radiotherapy inhibited the growth of bladder cancer tumor transplants and increased their radiosensitivity(P＜0.05).Conclusions LncRNA UCA1 is up-regulated in bladder cancer cells.Knockdown of LncRNA UCA1 can inhibit cell proliferation,induce apoptosis,and enhance the radiosensitivity of bladder cancer cells by up-regulating miR-520a-5p.
余洋;熊飞;陈晓波;孙伟;张炜
宜昌市中心人民医院泌尿外科,湖北宜昌 443000宜昌市中心人民医院泌尿外科,湖北宜昌 443000宜昌市中心人民医院泌尿外科,湖北宜昌 443000武汉市第三医院泌尿外科,武汉 430000武汉市第三医院泌尿外科,武汉 430000
医药卫生
LncRNA UCA1miR-520a-5p膀胱癌放疗敏感性增殖凋亡
LncRNA UCA1miR-520a-5pbladder cancerradiosensitivityproliferationapoptosis
《中国比较医学杂志》 2026 (8)
61-70,10
武汉市医学科研项目(WX21B04).
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