摘要
该文筛选了靶向抑制FTO的miRNA并探究其乳腺癌细胞生物学行为的影响。通过生物信息学的方法筛选出了影响乳腺癌患者生存的m6A去甲基化酶FTO后,再通过CCK-8实验验证了FTO的下调能够抑制乳腺癌细胞的增殖,随后预测出靶向FTO的miRNA-miR-504-5p,RT-qPCR和Western blot实验检测了乳腺癌细胞MCF-7和MDA-MB-231中miRNA-504-5p对FTO表达水平的影响,双荧光素酶报告基因实验验证了miRNA-504-5p与FTO的结合关系,采用CCK-8、Transwell小室实验、流式细胞术等探究了miRNA-504-5p mimic(类似物)通过调控FTO对乳腺癌细胞增殖、迁移、凋亡和细胞周期的影响。实验结果表明,低表达FTO的乳腺癌患者相较于高表达FTO的乳腺癌患者具有更高的生存概率,miRNA-504-5p作为潜在的靶向FTO的miRNA,能够在mRNA与蛋白水平抑制FTO的表达,并且miR-504-5p在FTO 3’-UTR的5 927–5 933位点处与FTO靶向结合,miR-504-5p能通过下调FTO抑制乳腺癌细胞的增殖与迁移并促进乳腺癌细胞的凋亡,使细胞阻滞在G0/G1期。综上所述,该研究发现了miR-504-5p能下调FTO的表达,抑制乳腺癌细胞的增殖与迁移,促进乳腺癌细胞的凋亡,使乳腺癌细胞的细胞周期阻滞。这可以为乳腺癌的分子机制探究与治疗提供潜在的参考价值。
This study aimed to identify miRNA that targeted FTO and its role in biological behaviors of breast cancer cells.Bioinformatic analysis was used to screen out the survival-related RNA methylation regulator FTO.CCK-8 assay was performed to evaluate the proliferation of breast cancer cells with FTO-downregulation.miR-504-5p was subsequently predicted as a potential miRNA targeting FTO.qRT-PCR and Western blot were applied to investigate the effects of miR-504-5p on the expression of FTO.Dual-luciferase reporter assay was utilized to validate the relationship between miR-504-5p and FTO.CCK-8 assay,Transwell assay and flow cytometry were performed to evaluate the influence of miR-504-5p on the proliferation,migration,apoptosis and cell cycle distribution of breast cancer cells.The results indicated that FTO low-expressed breast cancer patients had more favorable survival rate than the FTO high-expressed breast cancer patients.miR-504-5p was able to down-regulate the expression of FTO mRNA or protein and it binded with FTO in the position of 5 927–5 933 of FTO 3’-UTR.miR-504-5p attenuated the proliferation and migration,meanwhile promoted the apoptosis of breast cancer cells,blocked the cell cycles in G0/G1.In summary,this study found that miR-504-5p could down-regulate the expression of FTO,inhibit the proliferation and migration of breast cancer cells,promote the apoptosis of breast cancer cells,and block the cell cycle of breast cancer cells.This study may provide potential reference values for molecular mechanism study and therapy of breast cancer.
作者
陆双欣
淮瑞平
陈孟旸
李钢
杨慧
熊莉丽
LU Shuangxin;HUAI Ruiping;CHEN Mengyang;LI Gang;YANG Hui;XIONG Lili(School of Life Science and Engineering,Southwest Jiaotong University,Chengdu 610031,China)
出处
《中国细胞生物学学报》
CSCD
2021年第7期1418-1427,共10页
Chinese Journal of Cell Biology
基金
国家自然科学基金青年科学基金(批准号:31200999)
四川省科技计划项目(批准号:18YYJC0551)
中央高校基本科研业务费专项资金(批准号:2682020ZT112)资助的课题。
