Men Yingchao, Zhang Lei, Ai Hao
Department of Biochemistry and Molecular Biology, Jinzhou Medical University, Jinzhou 121000, China.
Department of Gynecology, Third Affiliated Hospital, Jinzhou Medical University, Jinzhou 121000, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2020 Jan 30;40(1):61-66. doi: 10.12122/j.issn.1673-4254.2020.01.10.
To investigate the role of microRNA-145-5p (miR-145-5p) in regulating the proliferation, migration, invasion and apoptosis of human endometrial carcinoma cells.
Human endometrial carcinoma Ishikawa cells were transfected with miR-145-5p mimic, miR-145-5p inhibitor, or their negative controls via liposome (Lipo2000), and the changes in the expression of miR-145-5p was verified by real-time PCR. The effects of overexpression or inhibition of miR-145-5p on the proliferation, migration, invasion and apoptosis of the cells were evaluated using MTT assay, wound healing assay, Transwell assay or flow cytometry. Bioinformatic analysis was performed to predict the target genes of miR-145-5p. The mRNA and protein expression levels of the downstream target of miR-145-5p, namely dual specific phosphatase 6 (DUSP6), were detected using real-time PCR and Western blotting.
Transfection of the cells with miR-145-5p mimic significantly suppressed the proliferation of Ishikawa cells, while transfection with miR-145-5p inhibitor obvious enhanced the proliferation of the cells ( < 0.05). Over-expression of miR-145-5p significantly suppressed the migration and invasion and promoted apoptosis of the cells, and inhibition of miR-145-5p caused the reverse changes ( < 0.05). Bioinformatic analysis showed that DUSP6 was the potential target gene of miR-145-5p. Over-expression of miR-145-5p significantly lowered while inhibition of miR-145-5p significantly enhanced the expression of DUSP6 protein ( < 0.05).
Overexpression of miR-145-5p inhibits the proliferation, migration and invasion and promotes apoptosis of endometrial cancer cells possibly by negative regulation of DUSP6 expression.
探讨微小RNA-145-5p(miR-145-5p)在调控人子宫内膜癌细胞增殖、迁移、侵袭及凋亡中的作用。
通过脂质体(Lipofectamine 2000)将miR-145-5p模拟物、miR-145-5p抑制剂或其阴性对照转染至人子宫内膜癌Ishikawa细胞,采用实时定量聚合酶链反应验证miR-145-5p表达的变化。运用MTT法、划痕实验、Transwell实验或流式细胞术评估miR-145-5p过表达或抑制对细胞增殖、迁移、侵袭及凋亡的影响。进行生物信息学分析以预测miR-145-5p的靶基因。采用实时定量聚合酶链反应和蛋白质免疫印迹法检测miR-145-5p下游靶标双特异性磷酸酶6(DUSP6)的mRNA和蛋白表达水平。
用miR-145-5p模拟物转染细胞显著抑制Ishikawa细胞的增殖,而用miR-145-5p抑制剂转染则明显增强细胞的增殖(P<0.05)。miR-145-5p过表达显著抑制细胞的迁移和侵袭并促进细胞凋亡,抑制miR-145-5p则导致相反的变化(P<0.05)。生物信息学分析表明DUSP6是miR-145-5p的潜在靶基因。miR-145-5p过表达显著降低而抑制miR-145-5p显著增强DUSP6蛋白的表达(P<0.05)。
miR-145-5p过表达可能通过负调控DUSP6表达来抑制子宫内膜癌细胞的增殖、迁移和侵袭并促进其凋亡。