P53介导miRNA-30e-5p抑制乳腺癌细胞恶性表型的实验研究  

Experimental Study of P53-Mediated MiRNA-30e-5p Inhibiting Malignant Phenotypes of Breast Cancer Cells

作　　者：刘嘉玉 李泽颖 陈玲燕 梅杰 马涛 张岩 Liu Jiayu;Li Zeying;Chen Lingyan;Mei Jie;Ma Tao;Zhang Yan(Department of Breast Surgery,Wuxi Maternal and Child Health Hospital,Wuxi School of Medicine of Jiangnan University,Wuxi 214122,Jiangsu,China;Wuxi Clinical Medical College,Nanjing Medical University,Wuxi 214023,Jiangsu,China)

机构地区：[1]江南大学无锡医学院无锡市妇幼保健院乳腺外科,江苏无锡214122 [2]南京医科大学无锡临床医学院,江苏无锡214023

出　　处：《肿瘤预防与治疗》2022年第11期979-986,共8页Journal of Cancer Control And Treatment

基　　金：江苏省妇幼保健项目(编号:F202034);无锡市双百人才基金(编号:BJ2020076);无锡市卫健委精准医学项目(编号:J202106);无锡市科技局重大项目(编号:N20201006)。

摘　　要：目的:探讨P53介导的miR-30e-5p对乳腺癌细胞恶性表型的抑制作用。方法:利用染色质免疫共沉淀试验与荧光定量PCR验证P53对miR-30e-5p的转录激活作用;分别采用CCK8试验、划痕试验和Transwell小室试验检测miR-30e-5p对于乳腺癌细胞增殖、迁移和侵袭能力的影响;通过激光共聚焦技术探究miR-30e-5p对β-catenin亚细胞定位的影响,并利用Western blot法检测β-catenin及其靶蛋白的表达情况。结果:P53可与MCF-7、MDAMB-453细胞的miR-30e-5p启动子区域结合,转录激活miR-30e-5p的表达(P <0.05);乳腺癌细胞转染miR-30e-5p后,其增殖、迁移、侵袭能力受到不同程度抑制(P <0.05);此外,miR-30e-5p可减少β-catenin的核内分布,抑制β-catenin靶基因表达。结论:P53可介导抑癌基因miR-30e-5p表达,通过抑制经典Wnt通路而抑制乳腺癌细胞恶性表型。Objective: To explore the inhibitory effect of p53-mediated miR-30e-5p on malignant phenotype of breast cancer cells. Methods: Chromatin immunoprecipitation and quantitative PCR were used to verify the transcriptional activation effect of P53 on miR-30e-5p. The effects of miR-30e-5p on the proliferation, migration and invasion of breast cancer cells were detected by CCK8, wound healing and Transwell assays, respectively. Confocal laser was used to explore the effect of miR-30e-5p on the subcellular localization of β-catenin. Western blot was used to detect the expressions of β-catenin and its target proteins. Results:P53 can bind to the miR-30e-5p promoter region of MCF-7 and MDA-MB-453 cells, and transcriptionally activates the expression of miR-30e-5p( P < 0.05). The breast cancer cells’ proliferation, migration and invasion abilities were inhibited to varying degrees after they were transfected with miR-30e-5p( P < 0.05). In addition, miR-30e-5p can reduce the distribution of β-catenin in nuclei and inhibit the expression of target genes of β-catenin. Conclusion : P53 can mediate the expression of miR-30e-5p, a tumor suppressor gene, and inhibit the malignant phenotypes of breast cancer cells by mediating the canonical Wnt pathway.

关 键 词：乳腺癌 P53 miR-30e-5p 经典Wnt通路 

分 类 号：R737.2[医药卫生—肿瘤]