School of Clinical Medicine, Ningxia Medical University, Yinchuan, China.
Ningxia Human Stem Cell Institute, General Hospital of Ningxia Medical University, Yinchuan, China.
Bioengineered. 2022 May;13(5):12350-12364. doi: 10.1080/21655979.2022.2074107.
In this study, we employed multiple laboratory techniques to acknowledge the biological activities and processes of Per2 and Id3 in glioma. We analyzed TCGA and CGGA databases for seeking association among Per2, Id3, and clinical features in glioma. Immunohistochemistry and Western blot were used to detect protein expression levels. CCK-8 assay, colony formation assay, Transwell assay, the wound healing assay, flow cytometric, and Xenograft nude mice were used to acknowledge the impact of Per2 and Id3 on biological behavior of glioma. The results showed that the Per2 mRNA expression was negatively correlated with the WHO grade, while the Id3 mRNA expression was positively correlated with the WHO grade in patients with glioma in TCGA and CGGA databases. Per2 and Id3 maintained separate prognostic abilities and had a negative connection in human glioma. In the clinical sample study, Per2 and Id3 were validated at the protein level with the same results compared to the mRNA expression level in TCGA and CGGA. By using a wide range of functional examples, overexpression of Per2 restrains malignant biological behaviors in glioma cells by many ways, while Id3 promotes malignant biological behaviors in glioma cells. Furthermore, overexpression of Per2 can inhibit Id3 expression via regulating PTEN/AKT/Smad5 signaling pathway and thereby abolish malignant biological behaviors that are caused by Id3 overexpression. These results suggested that Per2 inhibits glioma cell proliferation through regulating PTEN/AKT/Smad5/Id3 signaling pathway, which may be a viable therapeutic target for glioma.
在这项研究中，我们采用了多种实验室技术来确认 Per2 和 Id3 在神经胶质瘤中的生物学活性和过程。我们分析了 TCGA 和 CGGA 数据库，以寻找 Per2、Id3 与神经胶质瘤临床特征之间的关联。我们使用免疫组织化学和 Western blot 来检测蛋白表达水平。CCK-8 assay、集落形成 assay、Transwell assay、划痕愈合 assay、流式细胞术和裸鼠异种移植用于研究 Per2 和 Id3 对神经胶质瘤生物学行为的影响。结果表明，在 TCGA 和 CGGA 数据库中，Per2 mRNA 表达与 WHO 分级呈负相关，而 Id3 mRNA 表达与 WHO 分级呈正相关。Per2 和 Id3 在人类神经胶质瘤中具有独立的预后能力，且呈负相关。在临床样本研究中，与 TCGA 和 CGGA 的 mRNA 表达水平相比，Per2 和 Id3 在蛋白水平上得到了相同的验证。通过使用广泛的功能实例，过表达 Per2 通过多种方式抑制神经胶质瘤细胞的恶性生物学行为，而 Id3 则促进神经胶质瘤细胞的恶性生物学行为。此外，过表达 Per2 可以通过调节 PTEN/AKT/Smad5 信号通路抑制 Id3 表达，从而消除 Id3 过表达引起的恶性生物学行为。这些结果表明，Per2 通过调节 PTEN/AKT/Smad5/Id3 信号通路抑制神经胶质瘤细胞增殖，可能成为神经胶质瘤的一种可行的治疗靶点。
