miR-496通过SFMBT1抑制子宫颈癌HeLa细胞增殖、迁移和侵袭的相关研究

miR-496 inhibits proliferation,migration and invasion of HeLa cell in cervical cancer via SFMBT1

背景与目的:微小RNA(microRNA,miRNA)已被证实与子宫颈癌的发病相关。分析miR-496与SFMBT1的相互关系,阐明miR-496在子宫颈癌中的作用及其机制。方法:预测并验证miR-496的靶基因。实时荧光定量聚合酶链式反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)和蛋白质印迹法(Western blot)分析miR-496 mimics对SFMBT1表达的影响。使用免疫荧光和Western blot分析SFMBT1在子宫颈癌组织中的表达变化。细胞计数试剂盒-8(cell counting kit-8,CCK-8)法和transwell实验分别检测miR-496和SFMBT1对HeLa细胞增殖、迁移和侵袭的影响。观察miR-496在BALB/c裸小鼠移植瘤模型中对肿瘤生长的影响。结果:相比癌旁正常组织,SFMBT1在子宫颈癌组织中的mRNA(1.69±0.23 vs 1.00±0.12)以及蛋白(1.73±0.28 vs 1.00±0.15)均呈高表达,miR-496能通过特异性结合SFMBT1的3’-UTR抑制SFMBT1的mRNA(0.81±0.07 vs 1.00±0.15)以及蛋白表达(0.26±0.02 vs 1.00±0.14);细胞实验中相比对照组,miR-496 mimics处理组能抑制HeLa细胞增殖(1.39±0.10 vs 2.01±0.09)、迁移(40.50±3.17 vs 28.42±1.21)和侵袭能力(15.03±1.67 vs 25.71±2.56),但相比miR-496 mimics处理组,miR-496+SFMBT1处理组迁移(33.21±2.66 vs 19.28±1.50)和侵袭能力增强(30.11±2.73 vs 15.03±1.67)。子宫颈癌裸小鼠移植瘤模型发现,相比对照组miR-496组裸小鼠中移植瘤体积、重量均明显降低。miR-496组裸小鼠中病变程度、SFMBT1表达和Ki-67增殖指数均明显降低,差异有统计学意义(P<0.01)。结论:miR-496通过靶向调控SFMBT1抑制子宫颈癌细胞的生物学行为和裸小鼠移植瘤的生长。

Background and purpose:MicroRNA(miRNA)has been proved to be related to the pathogenesis of cervical cancer.The study aimed to analyze the mutual relation between miR-496 and SFMBT1 and the mechanism of miR-496 in cervical cancer.Methods:Target gene of miR-496 was predicted and confirmed.The effect of miR-496 mimics on SFMBT1 expression was detected by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)and Western blot.Changes of SFMBT1 expression in cervical cancer tissues were analyzed using immunofluorescence and Western blot.Cell counting kit-8(CCK-8)and transwell assays were used to detect the effects of both miR-496 and SFMBT1 on HeLa cell proliferation,migration and invasion,respectively.The impact of miR-496 on tumor growth was also explored in nude mice model.Results:Compared with para-cancerous tissues,mRNA(1.69±0.23 vs 1.00±0.12)and protein(1.73±0.28 vs 1.00±0.15)of SFMBT1 were highly expressed in tissues of cervical cancer;miR-496 could inhibit mRNA(0.81±0.07 vs 1.00±0.15)and protein(0.26±0.02 vs 1.00±0.14)expression of SFMBT1 via specifically binding to the 3’-UTR of SFMBT1.In cell experiments,HeLa cell proliferation(1.39±0.10 vs 2.01±0.09),migration(40.50±3.17±vs 28.42±1.21)and invasion(15.03±1.67 vs 25.71±2.56)abilities were suppressed in miR-496 mimics group compared with control group.Abilities of cell migration(33.21±2.66 vs 19.28±1.50)and invasion(30.11±2.73 vs 15.03±1.67)were enhanced in miR 496+SFMBT1 group compared with miR-496 mimics group.In nude mice xenograft model of cervical cancer,tumor volume and weight were obviously reduced in miR-496 group compared with control group.The degree of lesion and SFMBT1 expression and Ki-67 proliferative index were obviously decreased in miR-496 group(P<0.01).Conclusion:MiR-496 inhibits biological behaviors of cervical cancer cells and transplanted tumor growth of nude mice via targeted regulation of SFMBT1.