Liu Jing, Chen Weiyan, Zhang Haiyan, Liu Ting, Zhao Lin
Department of Gynecology, Linyi Tumor Hospital, Linyi, Shandong 276002, P.R. China.
Oncol Lett. 2017 Nov;14(5):5711-5718. doi: 10.3892/ol.2017.6953. Epub 2017 Sep 15.
The present study aimed to investigate the potential role of microRNA (miR)-214 in targeting the phosphatase and tensin homolog (PTEN)-mediated phosphoinositide 3-kinase (PI3K)/Akt signaling pathway in ovarian cancer (OC). The target gene of miR-214 was determined by luciferase reporter gene assay and was indicated to be PTEN. Human SK-OV-3 cells were transfected with a miR-214 inhibitor and a miR-214 mimic, and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to detect relative expression of miR-214. The MTT assay was performed to detect cell viability following transfection. Cell cycle and apoptosis were assessed by staining with propidium iodide (PI) and double staining with Annexin V/PI, respectively. The expression levels of PTEN and PI3K/Akt signaling pathway-associated proteins were detected by western blot analysis. The expression of miR-214 in tumor tissues and normal tissues was detected by RT-qPCR, and PTEN expression was detected by immunohistochemistry. SK-OV-3 cells transfected with a miR-214 inhibitor showed significantly inhibited cell viability and proliferation, and markedly increased apoptotic rate. SK-OV-3 cells transfected with miR-214 mimic showed significantly increased viability and proliferation, and markedly decreased apoptotic rate. The cells transfected with a miR-214 inhibitor exhibited significantly upregulated PTEN expression and significantly downregulated phosphatidylinositol (3,4,5)-trisphosphate (PIP3), phosphorylated (p)-Akt and p-glycogen synthase kinase (GSK)-3β expression. The cells transfected with miR-214 mimic exhibited significantly downregulated PTEN expression and significantly upregulated PIP3, p-Akt and p-GSK-3β expressions. The OC tissues exhibited an increased expression of miR-214 and a reduced positive rate of PTEN expression compared with adjacent normal tissues. miR-214 may activate the PI3K/Akt signaling pathway by downregulating the targeted PTEN, which may promote OC cell proliferation and inhibit apoptosis.
本研究旨在探讨微小RNA(miR)-214在卵巢癌(OC)中靶向磷酸酶和张力蛋白同源物(PTEN)介导的磷酸肌醇3激酶(PI3K)/Akt信号通路的潜在作用。通过荧光素酶报告基因检测确定miR-214的靶基因,结果表明为PTEN。用miR-214抑制剂和miR-214模拟物转染人SK-OV-3细胞,采用逆转录定量聚合酶链反应(RT-qPCR)检测miR-214的相对表达。转染后通过MTT法检测细胞活力。分别用碘化丙啶(PI)染色和Annexin V/PI双染法评估细胞周期和凋亡情况。通过蛋白质印迹分析检测PTEN和PI3K/Akt信号通路相关蛋白的表达水平。用RT-qPCR检测肿瘤组织和正常组织中miR-214的表达,用免疫组织化学检测PTEN表达。转染miR-214抑制剂的SK-OV-3细胞显示细胞活力和增殖受到显著抑制,凋亡率明显增加。转染miR-214模拟物的SK-OV-3细胞显示活力和增殖显著增加,凋亡率明显降低。转染miR-214抑制剂的细胞PTEN表达显著上调,磷脂酰肌醇(3,4,5)-三磷酸(PIP3)、磷酸化(p)-Akt和p-糖原合酶激酶(GSK)-3β表达显著下调。转染miR-214模拟物的细胞PTEN表达显著下调,PIP3、p-Akt和p-GSK-3β表达显著上调。与相邻正常组织相比,OC组织中miR-214表达增加,PTEN表达阳性率降低。miR-214可能通过下调靶向的PTEN激活PI3K/Akt信号通路,这可能促进OC细胞增殖并抑制凋亡。