Department of Pharmacy, Yunnan Hospital of Traditional Chinese Medicine, Kunming, Yunnan 650021, P.R. China.
Department of Pediatrics, Kunming Hospital of Traditional Chinese Medicine, Kunming, Yunnan 650011, P.R. China.
Mol Med Rep. 2021 Jun;23(6). doi: 10.3892/mmr.2021.12071. Epub 2021 Apr 13.
The present study aimed to analyze the effects and underlying mechanisms of microRNA (miR)‑29‑3p on the proliferation and secretory abilities of prolactinoma cells by targeting insulin‑like growth factor (IGF)‑1/β‑catenin. The relationship between miR‑29a‑3p and the survival of prolactinoma cells was analyzed with the Kaplan‑Meier method in reference to The Cancer Genome Atlas. The expression levels of miR‑29a‑3p and IGF‑1 in MMQ and GH3 cells were detected. A dual‑luciferase reporter gene assay was performed to verify the combination of miR‑29a‑3p and IGF‑1. Cells were transfected with a miR‑29a‑3p mimic and/or IGF‑1 pcDNA3.1 to analyze the effects on the proliferation, apoptosis and secretion of prolactin (PRL) and growth hormone (GH) of prolactinoma cells. The effects on β‑catenin in the cytoplasm and nucleus were investigated by western blot analysis. The results showed that miR‑29a‑3p expression was low in MMQ and GH3 cells. Overexpression miR‑29a‑3p inhibited IGF‑1 mRNA and protein expression. miR‑29a‑3p inhibited cell proliferation and PRL and GH expression, and promoted apoptosis by inhibiting IGF‑1. Increasing the expression of miR‑29a‑3p increased β‑catenin levels in the cytoplasm, whereas IGF‑1 promoted β‑catenin activation and entry into the nucleus, and reversed the inhibitory effects of miR‑29a‑3p on β‑catenin. To conclude, miR‑29a‑3p inhibited the proliferation and secretory abilities of prolactinoma cells by inhibiting nuclear translocation of β‑catenin via a molecular mechanism that is inseparable from IGF‑1.
本研究旨在分析微小 RNA(miR)-29-3p 通过靶向胰岛素样生长因子(IGF)-1/β-连环蛋白对泌乳素瘤细胞增殖和分泌能力的影响及其潜在机制。采用 Kaplan-Meier 方法结合 The Cancer Genome Atlas 分析 miR-29a-3p 与泌乳素瘤细胞生存的关系。检测 MMQ 和 GH3 细胞中 miR-29a-3p 和 IGF-1 的表达水平。通过双荧光素酶报告基因检测验证 miR-29a-3p 和 IGF-1 的结合。转染 miR-29a-3p 模拟物和/或 IGF-1 pcDNA3.1,分析其对泌乳素瘤细胞增殖、凋亡和泌乳素(PRL)和生长激素(GH)分泌的影响。采用 Western blot 分析检测细胞浆和细胞核中β-连环蛋白的变化。结果表明,MMQ 和 GH3 细胞中 miR-29a-3p 表达水平较低。过表达 miR-29a-3p 抑制 IGF-1 mRNA 和蛋白表达。miR-29a-3p 通过抑制 IGF-1 抑制细胞增殖及 PRL 和 GH 表达,并促进细胞凋亡。增加 miR-29a-3p 的表达增加了细胞浆中β-连环蛋白的水平,而 IGF-1 促进了β-连环蛋白的激活和核内转位,并逆转了 miR-29a-3p 对β-连环蛋白的抑制作用。综上所述,miR-29a-3p 通过抑制核内 β-连环蛋白的易位,通过一种与 IGF-1 密不可分的分子机制,抑制了泌乳素瘤细胞的增殖和分泌能力。