Department of Urology Surgery, Shigatse People's Hospital, Shigatse, Tibet, P.R. China.
Department of Gastric Cancer and Soft-Tissue Sarcoma Sugery, Fudan University Shanghai Cancer Center, Shanghai, P.R. China.
J Cell Physiol. 2018 Jan;234(1):464-474. doi: 10.1002/jcp.26591. Epub 2018 Jun 12.
This study was aimed to uncover the effects of miR-124-3p on bladder cancer (BC) by regulating DNA methyltransferase 3B. The expressions of miR-124-3p and DNMT3B mRNA in BC tissues and cell lines were detected using RT-PCR. The expression of DNMT3B in cells was determined using Western blot and immunohistochemistry in tissues. In addition, chromogenic in situ hybridization staining was used to measure the expression of miR-124-3p in tissues. BC cells were transfected with miR-124-3p mimics, miR-124-3p inhibitors, DNMT3B siRNAs, and DNMT3B cDNAs + miR-124-3p mimics. Subsequently, cell proliferation, apoptosis, migration, and invasion were measured using CCK-8, the cytometry test, wound healing assay, and Transwell assay, respectively. Finally, the relationship between miR-124-3p and DNMT3B was confirmed using dual luciferase reporter gene assay. MiR-124-3p expression was significantly lower and the level of DNMT3B was significantly higher in BC tissues and cell lines compared with the normal controls. MiR-124-3p was verified to target DNMT3B. The transfection of miR-124-3p mimics and DNMT3B siRNAs down-regulated BC cell proliferation, migration, and invasion, as well as induced cell apoptosis; miR-124-3p inhibitors promoted BC cell proliferation, migration, invasion, and reduced cell apoptosis; and the effects of DNMT3B cDNAs can be compromised by miR-124-3p mimics. Thus, we concluded that miR-124-3p could suppress the proliferation, migration, invasion, and promote apoptosis of BC cells by targeting DNMT3B.
本研究旨在通过调控 DNA 甲基转移酶 3B（DNMT3B）来揭示 miR-124-3p 对膀胱癌（BC）的影响。采用 RT-PCR 检测 BC 组织和细胞系中 miR-124-3p 和 DNMT3B mRNA 的表达。采用 Western blot 和免疫组织化学检测组织中 DNMT3B 的表达。此外，采用显色原位杂交染色检测组织中 miR-124-3p 的表达。将 miR-124-3p 模拟物、miR-124-3p 抑制剂、DNMT3B siRNAs 和 DNMT3B cDNA+miR-124-3p 模拟物转染至 BC 细胞中。随后，采用 CCK-8 法、细胞计数法、划痕愈合试验和 Transwell 试验分别测量细胞增殖、凋亡、迁移和侵袭。最后，采用双荧光素酶报告基因试验证实 miR-124-3p 与 DNMT3B 之间的关系。与正常对照组相比，BC 组织和细胞系中 miR-124-3p 的表达显著降低，DNMT3B 的水平显著升高。证实 miR-124-3p 可靶向作用于 DNMT3B。转染 miR-124-3p 模拟物和 DNMT3B siRNAs 可下调 BC 细胞增殖、迁移和侵袭，并诱导细胞凋亡；miR-124-3p 抑制剂促进 BC 细胞增殖、迁移、侵袭，并减少细胞凋亡；而 miR-124-3p 模拟物可削弱 DNMT3B cDNA 的作用。因此，我们得出结论，miR-124-3p 可通过靶向作用于 DNMT3B 抑制 BC 细胞的增殖、迁移、侵袭并促进细胞凋亡。
