Department of Obstetrics and Gynecology, Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China.
Department of Pharmacy, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China.
Mol Med Rep. 2017 Nov;16(5):7267-7276. doi: 10.3892/mmr.2017.7523. Epub 2017 Sep 19.
Integrin‑linked kinase (ILK) is overexpressed in ovarian cancer (OC), and ILK gene silencing results in apoptosis in OC cells. In the present study, the mechanism by which ILK induces apoptosis was explored from the perspective of microRNA (miRNA) expression. Alterations in the global miRNA expression profile were detected using a miRNA microarray after OC cells were transduced with an ILK small hairpin RNA lentivirus. ILK silencing led to a significant upregulation of 14 miRNAs by at least 1.5‑fold. These findings were validated by reverse transcription‑quantitative polymerase chain reaction. A pathway analysis of experimentally validated target genes revealed the inhibition of multiple cancer‑associated signaling pathways and the wnt signaling pathway. Compared with cells transfected with scrambled RNA, the ILK‑silenced cells had remarkably lower expression of wnt ligands (wnt3a, wnt4 and wnt5a) and downstream β‑catenin. ILK silencing led to apoptosis of OC cells and impaired the migratory ability. Taken together, the present results suggested that miRNA‑mediated wnt pathway alterations are involved in the anti‑apoptotic role of ILK in OC. It was also indicated that ILK silencing reduced the ability of OC cells to adhere to fibronectin, which may lead to unstable focal contact. Consistently, the phosphorylation levels of focal adhesion kinase and RAC‑α serine/threonine protein kinase were downregulated. The present work demonstrated the first global miRNA expression profile of OC cells when ILK was inhibited, and this expression profile may provide a basis for the development of biomarkers and therapeutic targets for OC.
整合素连接激酶（ILK）在卵巢癌（OC）中过表达，ILK 基因沉默会导致 OC 细胞凋亡。在本研究中，从 miRNA（miRNA）表达的角度探讨了 ILK 诱导凋亡的机制。用 ILK 短发夹 RNA 慢病毒转导 OC 细胞后，用 miRNA 微阵列检测全局 miRNA 表达谱的变化。ILK 沉默导致至少 1.5 倍的 14 个 miRNA 显著上调。通过逆转录-定量聚合酶链反应验证了这些发现。对实验验证的靶基因的通路分析显示，多个癌症相关信号通路和 wnt 信号通路被抑制。与转染 scrambled RNA 的细胞相比，ILK 沉默的细胞中 wnt 配体（wnt3a、wnt4 和 wnt5a）和下游 β-连环蛋白的表达显著降低。ILK 沉默导致 OC 细胞凋亡并损害迁移能力。综上所述，本研究结果表明 miRNA 介导的 wnt 通路改变参与了 ILK 在 OC 中的抗凋亡作用。研究还表明，ILK 沉默降低了 OC 细胞黏附纤维连接蛋白的能力，这可能导致不稳定的焦点接触。一致地，粘着斑激酶和 RAC-α 丝氨酸/苏氨酸蛋白激酶的磷酸化水平下调。本研究首次展示了 ILK 抑制时 OC 细胞的全局 miRNA 表达谱，该表达谱可能为 OC 的生物标志物和治疗靶点的开发提供基础。
