MicroRNA-126-3p suppresses HeLa cell proliferation, migration and invasion, and increases apoptosis via the PI3K/PDK1/AKT pathway
MicroRNA-126-3p suppresses HeLa cell proliferation, migration and invasion, and increases apoptosis via the PI3K/PDK1/AKT pathway
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DOI：
10.3892/or.2020.7512
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发表时间：
2020-04-01
期刊：
影响因子：
4.2
通讯作者：
Fujii, Takuma
中科院分区：
文献类型：
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作者：
Ichikawa, Ryoko;Kawasaki, Rie;Fujii, Takuma
我们之前报道，相对于正常宫颈粘液，微小RNA 126-3p (miR-126-3p) 在患有明显宫颈癌或癌前病变的患者宫颈粘液中的含量显着更高。在这里，我们研究了宫颈癌细胞系 HeLa 中强制表达 miR-126-3p 对增殖、迁移、侵袭、凋亡和蛋白表达的影响。我们用 miR-126-3p miRNA 转染 HeLa 细胞，发现与转染阴性对照模拟物的细胞相比，通过细胞计数、伤口愈合、细胞迁移和侵袭测定，这些细胞的增殖、迁移和侵袭显着减少。 miR-126-3p转染细胞中磷酸肌醇3激酶（PI3K）、磷酸化3-磷酸肌醇依赖性蛋白激酶1（p-PDK1）和p-AKT蛋白的水平较低。磷酸化 70S6K (p-p70S6K)、磷酸化糖原合酶激酶 3 beta (p-GSK3 beta)、磷酸化 S6K (p-S6K)、细胞周期蛋白 D1、磷酸化 p21 激活激酶 1 (p-PAK1)、Rho 相关卷曲螺旋蛋白激酶 1 (ROCK1)、强直性肌营养不良相关 CDC42 结合激酶α (MRCK α) 和磷脂酶 C γ 1 (p-PLC γ 1) 也下调。这表明 PI3K/PDK1/AKT 通路的下游效应器是 miR-126-3p 抑制的靶标。相比之下，细胞凋亡相关蛋白，包括 BCL-2 相关细胞死亡激动剂 (Bad)、B 细胞超大淋巴瘤 (Bcl-xL) 和 BCL-2 相关 X (Bax)，均被 miR-126-3p 上调，导致 caspase 3/7 活性增加和细胞凋亡。因此，miR-126-3p的强制表达抑制细胞迁移和侵袭，并通过调节HeLa细胞中的PI3K/PDK1/AKT通路诱导细胞凋亡。因此，高水平的miR-126-3p可能抑制宫颈癌的发生，通过miR-126-3p靶向PI3K/PDK1/AKT通路可能代表治疗宫颈癌患者的新方法。
We previously reported that relative to normal cervical mucus, microRNA 126-3p (miR-126-3p) is present in significantly greater amounts in the cervical mucus of patients with overt cervical cancer or precursor lesions. Here, we investigated the effects of enforced miR-126-3p expression in the cervical cancer cell line, HeLa, on proliferation, migration, invasion, apoptosis and protein expression. We transfected HeLa cells with miR-126-3p miRNA and found that proliferation, migration and invasion by cell counting, wound healing, cell migration and invasion assay were significantly reduced in these cells relative to those transfected with a negative control mimic. The levels of phosphoinositide 3 kinase (PI3K), phosphorylated 3-phosphoinositide-dependent protein kinase-1 (p-PDK1) and p-AKT proteins were lower in the miR-126-3p-transfected cells. Phosphorylated 70S6K (p-p70S6K), phosphorylated glycogen synthase kinase 3 beta (p-GSK3 beta), phosphorylated S6K (p-S6K), cyclin D1, phosphorylated p21-activated kinase 1 (p-PAK1), Rho associated coiled-coil containing protein kinase 1 (ROCK1), myotonic dystrophy-related CDC42-binding kinases alpha (MRCK alpha) and phospholipase C gamma 1 (p-PLC gamma 1) were also downregulated. This suggests that downstream effectors of the PI3K/PDK1/AKT pathway are targets for inhibition by miR-126-3p. In contrast, apoptotic-related proteins including the BCL-2-associated agonist of cell death (Bad), B-cell lymphoma-extra-large (Bcl-xL) and BCL-2-associated X (Bax), were all upregulated by miR-126-3p, resulting in increased caspase 3/7 activity and apoptosis. Thus, enforced expression of miR-126-3p inhibited cell migration and invasion and also induced apoptosis by regulating the PI3K/PDK1/AKT pathway in HeLa cells. Hence, high levels of miR-126-3p may inhibit cervical carcinogenesis, and targeting the PI3K/PDK1/AKT pathway via miR-126-3p could represent a new approach for treating patients with cervical cancer.