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

doi:10.19401/j.cnki.1007-3639.2021.08.002

中国癌症杂志 ›› 2021, Vol. 31 ›› Issue (8): 697-703.doi: 10.19401/j.cnki.1007-3639.2021.08.002

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

方艳惠 ¹ ，马彩娟 ² ，张纯丽 ¹ ，翟嘉威 ¹ ，朱巧英 ¹ ，刘茵 ¹ ，赵喜娃 ¹

1. 河北医科大学第四医院东院区妇科，河北石家庄 050011 ；
2. 晋州同济医院妇产科，河北石家庄 052260

出版日期:2021-08-30 发布日期:2021-09-03
通信作者: 赵喜娃 E-mail: yyffh01@163.com
基金资助:
河北省2021年度医学科学研究重点课题（20210082）。

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

FANG Yanhui ¹ , MA Caijuan ² , ZHANG Chunli ¹ , ZHAI Jiawei ¹ , ZHU Qiaoying ¹ , LIU Yin ¹ , ZHAO Xiwa ¹

1. Department of Gynecology, The Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei Province, China; 2. Department of Obstetrics and Gynecology, Jinzhou Tongji Hospital, Shijiazhuang 052260, Hebei Province, China

Published:2021-08-30 Online:2021-09-03
Contact: ZHAO Xiwa E-mail: yyffh01@163.com

摘要/Abstract

摘要： 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。

关键词: SFMBT1, miR-496, 子宫颈癌, HeLa

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 (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.

Key words: SFMBT1, miR-496, Cervical cancer, HeLa

方艳惠, 马彩娟, 张纯丽, 翟嘉威, 朱巧英, 刘茵, 赵喜娃. 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[J]. 中国癌症杂志, 2021, 31(8): 697-703.

FANG Yanhui , MA Caijuan , ZHANG Chunli , ZHAI Jiawei , ZHU Qiaoying , LIU Yin , ZHAO Xiwa . miR-496 inhibits proliferation, migration and invasion of HeLa cell in cervical cancer via SFMBT1[J]. China Oncology, 2021, 31(8): 697-703.

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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

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