Department of Pediatric Surgery, Zigong First People's Hospital, Zigong, Sichuan 643099, P.R. China.
Department of Clinical, Chongqing Jiulongpo District Hospital of Traditional Chinese Medicine, Chongqing 400039, P.R. China.
Mol Med Rep. 2022 Jun;25(6). doi: 10.3892/mmr.2022.12721. Epub 2022 Apr 29.
The transcription factor homeobox protein Hox‑B2 (HOXB2) and its downstream factor nucleolar and spindle‑associated protein 1 (NUSAP1) play important regulatory roles in cell proliferation, invasion and migration. However, their effects and specific mechanisms in nephroblastoma have not been previously investigated, to the best of our knowledge. Therefore, in the present study, the mRNA and protein expression levels of HOXB2 and NUSAP1 were determined in nephroblastoma cells using reverse transcription‑quantitative PCR and western blot analyses, respectively. Furthermore, cell transfection experiments were carried out to knock down NUSAP1 and overexpress HOXB2 in nephroblastoma cell lines. The proliferative, invasive and migratory abilities of nephroblastoma cells were assessed by MTT, EdU, colony formation, wound healing and Transwell assays. In addition, the JASPAR website was used to predict the association between HOXB2 and NUSAP1, which was further verified by dual‑luciferase reporter and chromatin immunoprecipitation assays. Finally, the expression levels of the PI3K/Akt signaling pathway‑related proteins were measured by western blot analysis. The results showed that the expression of NUSAP1 was abnormally upregulated in nephroblastoma cell lines. However, NUSAP1 silencing attenuated the proliferation, invasion and migration abilities of nephroblastoma cells. The results also suggested that HOXB2 could transcriptionally activate NUSAP1. Therefore, HOXB2 overexpression abrogated the inhibitory effect of NUSAP1 silencing on the proliferation and metastasis of nephroblastoma cells, possibly via the PI3K/Akt signaling pathway. The aforementioned findings indicated that HOXB2 may upregulate NUSAP1 to promote the proliferation, invasion and migration of nephroblastoma cells via the PI3K/Akt signaling pathway.
转录因子同源盒蛋白 Hox-B2（HOXB2）及其下游因子核仁纺锤体相关蛋白 1（NUSAP1）在细胞增殖、侵袭和迁移中发挥重要的调节作用。然而，据我们所知，它们在肾母细胞瘤中的作用和具体机制尚未得到研究。因此，在本研究中，分别采用逆转录-定量 PCR 和 Western blot 分析检测 HOXB2 和 NUSAP1 在肾母细胞瘤细胞中的 mRNA 和蛋白表达水平。此外，通过细胞转染实验敲低 NUSAP1 并过表达 HOXB2 在肾母细胞瘤细胞系中。通过 MTT、EdU、集落形成、划痕愈合和 Transwell 分析评估肾母细胞瘤细胞的增殖、侵袭和迁移能力。此外，使用 JASPAR 网站预测 HOXB2 和 NUSAP1 之间的关联，通过双荧光素酶报告基因和染色质免疫沉淀实验进一步验证。最后，通过 Western blot 分析测定 PI3K/Akt 信号通路相关蛋白的表达水平。结果显示，NUSAP1 在肾母细胞瘤细胞系中的表达异常上调。然而，沉默 NUSAP1 可减弱肾母细胞瘤细胞的增殖、侵袭和迁移能力。结果还表明，HOXB2 可转录激活 NUSAP1。因此，过表达 HOXB2 可消除 NUSAP1 沉默对肾母细胞瘤细胞增殖和转移的抑制作用，可能通过 PI3K/Akt 信号通路。上述研究结果表明，HOXB2 可能通过 PI3K/Akt 信号通路上调 NUSAP1 促进肾母细胞瘤细胞的增殖、侵袭和迁移。
