Wang Yufeng, Jie Luo, Gong Haifeng, Li Yuqin, Xie Anxia, Li Yanjun, Guo Hong
Department of Gynaecology and Obstetrics, Qinghai Provincial People's Hospital, Xining, Qinghai 810007, P.R. China.
Exp Ther Med. 2020 Aug;20(2):1379-1384. doi: 10.3892/etm.2020.8866. Epub 2020 Jun 10.
Effect of micro ribonucleic acid (miR)-30 on the proliferation of trophoblasts in preeclampsia (PE) rats through the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathway was studied. The miR-30 mimic was transfected into the trophoblast HTR8/SVNEO cell lines. The effects of expression level of miR-30 on the proliferation and hypoxia-induced apoptosis of HTR8/SVNEO cells were detected via methyl thiazolyl tetrazolium (MTT) assay and Annexin V/propidium iodide staining, respectively, using the flow cytometer. A total of 30 pregnant Sprague-Dawley rats were randomly divided into control group (CTL group, n=10), PE rat group (PE group, n=10) and PE + miR-30 Mimic group (PE+agomiR-30 group, n=10) using a random number table. The protein expression levels of phosphorylated ERK (p-ERK)1/2, ERK1/2, proliferating cell nuclear antigen (PCNA) and tubulin were determined using western blot analysis, and the mRNA expression level of ERK1/2 was detected via reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The expression level of PCNA in tissues was detected via immunohistochemistry. The results of MTT assay showed that the proliferation of HTR8/SVNEO cells significantly declined in hypoxic environment, while miR-30 promoted the proliferation of HTR8/SVNEO cells and alleviated the hypoxia-induced inhibition on cell proliferation. It was found that the trophoblast apoptosis rate was increased in hypoxia group compared with that in CTL group, while it was significantly decreased in miR-30 Mimic group compared with that in hypoxia group. PE group had obviously decreased p-ERK and PCNA expression levels as well as p-ERK/ERK ratio in placental tissues compared with CTL group, while PE+agomiR-30 group had an obviously increased expression level of PCNA as well as p-ERK/ERK ratio in placental tissues compared with PE group. MiR-30 activates the MAPK/ERK signaling pathway and increases the expression level of PCNA through raising the p-ERK level and p-ERK/ERK ratio, thereby inhibiting cell apoptosis and promoting cell proliferation.
研究了微小核糖核酸（miR）-30通过丝裂原活化蛋白激酶（MAPK）/细胞外信号调节激酶（ERK）途径对子痫前期（PE）大鼠滋养细胞增殖的影响。将miR-30模拟物转染到滋养细胞HTR8/SVNEO细胞系中。分别采用甲基噻唑基四氮唑（MTT）法和膜联蛋白V/碘化丙啶染色，利用流式细胞仪检测miR-30表达水平对HTR8/SVNEO细胞增殖和缺氧诱导凋亡的影响。采用随机数字表法将30只妊娠Sprague-Dawley大鼠随机分为对照组（CTL组，n = 10）、PE大鼠组（PE组，n = 10）和PE + miR-30模拟物组（PE+agomiR-30组，n = 10）。采用蛋白质免疫印迹分析测定磷酸化ERK（p-ERK）1/2、ERK1/2、增殖细胞核抗原（PCNA）和微管蛋白的蛋白表达水平，通过逆转录-定量聚合酶链反应（RT-qPCR）检测ERK1/2的mRNA表达水平。采用免疫组织化学法检测组织中PCNA的表达水平。MTT法结果显示，缺氧环境下HTR8/SVNEO细胞的增殖显著下降，而miR-30促进HTR8/SVNEO细胞的增殖，并减轻缺氧诱导的细胞增殖抑制。结果发现，缺氧组的滋养细胞凋亡率高于CTL组，而miR-30模拟物组的凋亡率明显低于缺氧组。与CTL组相比，PE组胎盘组织中p-ERK和PCNA表达水平以及p-ERK/ERK比值明显降低，而与PE组相比，PE+agomiR-30组胎盘组织中PCNA表达水平以及p-ERK/ERK比值明显升高。MiR-30通过提高p-ERK水平和p-ERK/ERK比值激活MAPK/ERK信号通路并增加PCNA表达水平，从而抑制细胞凋亡并促进细胞增殖。
