Department of Urology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou City, Henan Province, China.
Department of Urology, The Central Hospital of Xinyang City, Xinyang City, Henan Province, China.
Dis Markers. 2022 Aug 1;2022:8151161. doi: 10.1155/2022/8151161. eCollection 2022.
The effects of microRNA-29a-3p in the proliferation process of nerve cells are unclear. The purpose of this study is to delve into the regulatory role of microRNA-29a-3p, via interaction with phosphatase and tension homolog (PTEN), in the SH-SY5Y cell proliferation process. Different expressions of microRNA-29a-3p in the SH-SY5Y cells were constructed by transfected miRNA-29a-3p mimic and inhibitor. The effects of cell transfection and the mRNA expressions of PTEN, Akt, and mTOR were detected by qPCR. The expressions of PTEN, Akt, and mTOR protein and the phosphorylation levels of Akt and mTOR were examined using Western blotting. Nerve cell proliferation activity and neurite length of each group were measured and examined by the use of 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2Htetrazolium bromide (MTT), and morphological examination. We observed that the levels of PTEN mRNA and protein were distinctly decreased in the microRNA-29a-3p mimic group, but the expressions of the phosphorylated Akt and mTOR mRNA and protein were distinctly upregulated. In the transfected miRNA-29a-3p inhibitor SH-SY5Y cells, the expressions of miRNA-29a-3p were significantly suppressed; however, the expressions of PTEN gene and protein were significantly enhanced. The expressions of phosphorylated Akt and mTOR in the downregulated microRNA-29a-3p group distinctly were suppressed. The SH-SY5Y cell proliferation activity and neurite length in the upregulated microRNA-29a-3p group increased significantly. Our findings revealed that microRNA-29a-3p could enhance the proliferation activity of SH-SY5Y cells and promote neurite growth by inhibiting the expression of PTEN and regulating PI3K/Akt/mTOR signaling pathway.
miRNA-29a-3p 在神经细胞增殖过程中的作用尚不清楚。本研究旨在通过 miRNA-29a-3p 模拟物和抑制剂转染，深入研究 miRNA-29a-3p 与磷酸酶和张力蛋白同源物（PTEN）相互作用在 SH-SY5Y 细胞增殖过程中的调节作用。构建了 miRNA-29a-3p 模拟物和抑制剂转染后 SH-SY5Y 细胞中不同表达水平的 miRNA-29a-3p。通过 qPCR 检测细胞转染和 PTEN、Akt、mTOR 的 mRNA 表达。Western blot 检测 PTEN、Akt、mTOR 蛋白表达及 Akt 和 mTOR 的磷酸化水平。采用 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2H-四唑溴盐（MTT）检测各组神经细胞增殖活性和神经突长度，形态学观察。结果发现，miRNA-29a-3p 模拟物组中 PTEN mRNA 和蛋白水平明显降低，而磷酸化 Akt 和 mTOR mRNA 和蛋白表达明显上调。在转染 miRNA-29a-3p 抑制剂的 SH-SY5Y 细胞中，miRNA-29a-3p 的表达明显受到抑制，但 PTEN 基因和蛋白的表达明显增强。下调 miRNA-29a-3p 组磷酸化 Akt 和 mTOR 的表达明显受到抑制。上调 miRNA-29a-3p 组 SH-SY5Y 细胞增殖活性和神经突长度明显增加。我们的研究结果表明，miRNA-29a-3p 通过抑制 PTEN 的表达，调节 PI3K/Akt/mTOR 信号通路，增强 SH-SY5Y 细胞的增殖活性，促进神经突生长。
