miR-214 targets the PTEN-mediated PI3K/Akt signaling pathway and regulates cell proliferation and apoptosis in ovarian cancer.
miR-214 targets the PTEN-mediated PI3K/Akt signaling pathway and regulates cell proliferation and apoptosis in ovarian cancer.
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DOI：
10.3892/ol.2017.6953
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发表时间：
2017-11
期刊：
影响因子：
2.9
通讯作者：
Zhao L
中科院分区：
文献类型：
--
作者：
Liu J;Chen W;Zhang H;Liu T;Zhao L
本研究旨在探讨microRNA (miR)-214在卵巢癌（OC）中靶向磷酸酶和紧张素同源物（PTEN）介导的PI3K /Akt信号通路中的潜在作用。通过荧光素酶报告基因试验确定miR-214的靶基因，并提示为PTEN。用miR-214抑制剂和miR-214模拟物转染人SK-OV-3细胞，使用逆转录-定量聚合酶链反应（RT-qPCR）检测miR-214的相对表达。MTT法检测转染后的细胞活力。采用碘化丙啶（PI）染色法和Annexin V/PI双染色法观察细胞周期和凋亡情况。western blot检测PTEN和PI3K/Akt信号通路相关蛋白的表达水平。RT-qPCR检测miR-214在肿瘤组织和正常组织中的表达，免疫组织化学检测PTEN的表达。转染miR-214抑制剂的SK-OV-3细胞的细胞活力和增殖明显受到抑制，凋亡率明显升高。转染miR-214 mimic后，SK-OV-3细胞的活力和增殖能力明显增强，凋亡率明显降低。转染miR-214抑制剂的细胞显示PTEN表达显著上调，磷脂酰肌醇-三磷酸（PIP3）、磷酸化(p)-Akt和p-糖原合成酶激酶(GSK)-3β表达显著下调。转染miR-214 mimic的细胞显示PTEN表达显著下调，PIP3、p-Akt和p-GSK-3β表达显著上调。与邻近正常组织相比，OC组织miR-214表达增加，PTEN表达阳性率降低。miR-214可能通过下调靶向PTEN激活PI3K/Akt信号通路，从而促进OC细胞增殖，抑制细胞凋亡。
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 -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.
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