Department of Gynaecology and Obstetrics, The First Affiliated Hospital of Hunan Normal University.
Department of Anesthesiology, The Third Xiangya Hospital, Central South University, Changsha, People's Republic of China.
Anticancer Drugs. 2019 Jul;30(6):579-587. doi: 10.1097/CAD.0000000000000743.
Cervical cancer is the second most common gynecological malignancy worldwide and the tumorigenesis mechanisms of cervical cancer are still unclear. This study aimed to reveal the role of miR-29a in cervical cancer. The expression level of miR-29a and CDC42 was measured using qRT-PCR. Cell proliferation, apoptosis, migration, and invasion were detected using colony formation, flow cytometry analysis, wound-healing assay, and Transwell assay, respectively. Luciferase reporter assay was used to determine the binding of miR-29a with CDC42. CDC42/p21-activated kinase 1 (PAK1) pathway-related proteins were measured by western blotting. MiR-29a was downregulated and CDC42 was upregulated in cervical cancer cells. Luciferase reporter assay showed that miR-29a negatively regulated the expression of CDC42 by directly targeting 3'-UTR of CDC42. Cell proliferation, migration, and invasion were markedly inhibited, whereas cell apoptosis was significantly increased in Hela and CaSki cells transfected with miR-29a mimics. These effects were partly recovered by CDC42 overexpression. Protein levels of PAK1, p-PAK1, p-LIMK, and p-cofilin were significantly downregulated by miR-29a mimics, which was reversed by CDC42 overexpression and was increased by the miR-29a inhibitor. MiR-29a inhibited cell proliferation, migration, and invasion, as well as promoted cell apoptosis through repressing the PAK1/LIMK signaling pathway by targeting CDC42 in cervical cancer.
宫颈癌是全球第二大常见的妇科恶性肿瘤，其肿瘤发生机制仍不清楚。本研究旨在揭示 miR-29a 在宫颈癌中的作用。采用 qRT-PCR 检测 miR-29a 和 CDC42 的表达水平。通过集落形成、流式细胞术分析、划痕愈合试验和 Transwell 试验分别检测细胞增殖、凋亡、迁移和侵袭。采用荧光素酶报告实验确定 miR-29a 与 CDC42 的结合。通过 Western blot 检测 CDC42/p21 激活激酶 1（PAK1）通路相关蛋白。结果显示，宫颈癌细胞中 miR-29a 下调，CDC42 上调。荧光素酶报告实验表明，miR-29a 通过直接靶向 CDC42 的 3'-UTR 负调控 CDC42 的表达。转染 miR-29a 模拟物的 Hela 和 CaSki 细胞中，细胞增殖、迁移和侵袭明显受到抑制，而细胞凋亡显著增加。CDC42 的过表达部分恢复了这些效应。miR-29a 模拟物显著下调 PAK1、p-PAK1、p-LIMK 和 p-cofilin 的蛋白水平，而 CDC42 的过表达逆转了这一效应，miR-29a 抑制剂则增加了这一效应。miR-29a 通过靶向 CDC42 抑制 PAK1/LIMK 信号通路，抑制细胞增殖、迁移和侵袭，并促进细胞凋亡，从而抑制宫颈癌的发生。
