方艳惠, 马彩娟, 张纯丽, et al. miR-496 inhibits proliferation, migration and invasion of HeLa cell in cervical cancer via SFMBT1[J]. China Oncology, 2021, 31(8): 697-703.
方艳惠, 马彩娟, 张纯丽, et al. miR-496 inhibits proliferation, migration and invasion of HeLa cell in cervical cancer via SFMBT1[J]. China Oncology, 2021, 31(8): 697-703. DOI: 10.19401/j.cnki.1007-3639.2021.08.002.
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(P0.01)。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。
Abstract
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 (P0.01). Conclusion: MiR-496 inhibits biological behaviors of cervical cancer cells and transplanted tumor growth of nude mice via targeted regulation of SFMBT1.