Abstract：
Here, we explored the role of Prolyl 4-Hydroxylase Subunit Alpha 3 (P4HA3), the most recently identified member of the prolyl-4-hydroxylase (P4H) family, in head and neck squamous cell carcinoma (HNSCC) progression. P4HA3 is upregulated during cancer progression; however, its specific role in HNSCC progression remains elusive. Thus, this study aimed to elucidate the regulatory function of P4HA3 in HNSCC development and progression and to describe the underlying mechanisms. Initially, we analyzed the correlation between the expression of P4HA3 and the WNT pathway genes and clinicopathologic features in HNSCC based on microarray data from The Cancer Genome Atlas (TCGA). Next, we used Gene Oncology (GO) functional data to describe several potentially associated pathways in HNSCC. Then, we knocked down P4HA3 in SCC15 and SCC25 cells, two classic HNSCC cell lines, and assessed the resulting changes using RT-qPCR. Furthermore, we used Western blot to evaluate the regulatory role of P4HA3 in the epithelial-to-mesenchymal transition (EMT) and the WNT/β-catenin signaling pathway. To explore the effect of P4HA3 knockdown on tumor progression, in vivo experiments were conducted using a murine model. Immunohistochemistry assays were then employed to identify proteins associated with EMT and the WNT/β-catenin signaling pathway in tumor tissues. Upregulated P4HA3 in HNSCC patient tumor tissues was positively correlated with poor prognosis. Notably, P4HA3 knockdown significantly inhibited the proliferative and invasive abilities of HNSCC. The levels of genes and proteins associated with EMT and the WNT/β-catenin signaling pathway were also markedly reduced by P4HA3 knockdown. Importantly, the in vivo experiments demonstrated that P4HA3 can promote subcutaneous tumorigenesis in nude mice and knockdown of P4HA3 induce a significant ihibitation of EMT and WNT/β-catenin pathway detected by immunohistochemistry assay in tumor tissues. In summary, we demonstrated that P4HA3 is a promising diagnostic and therapeutic biomarker for HNSCC. As an oncogene, P4HA3 increases HNSCC proliferation by inducing the EMT and activating the WNT/β-catenin signaling pathway.
摘要：
这里,我们探索了脯氨酸4-羟化酶亚基Alpha3（P4HA3）的作用，最近发现的脯氨酸-4-羟化酶（P4H）家族的成员，头颈部鳞状细胞癌（HNSCC）进展。P4HA3在癌症进展期间上调;然而，其在HNSCC进展中的具体作用仍然难以捉摸。因此，本研究旨在阐明P4HA3在HNSCC发生和发展中的调控功能，并描述其潜在机制。最初，我们基于癌症基因组图谱(TCGA)的微阵列数据分析了HNSCC中P4HA3和WNT通路基因的表达与临床病理特征之间的相关性.接下来,我们使用基因肿瘤学（GO）功能数据来描述HNSCC中的几种潜在相关通路。然后，我们在SCC15和SCC25细胞中敲除P4HA3，两种经典的HNSCC细胞系，并使用RT-qPCR评估所得的变化。此外，我们使用Westernblot评估了P4HA3在上皮-间质转化(EMT)和WNT/β-catenin信号通路中的调节作用.探讨P4HA3基因敲低对肿瘤进展的影响,使用小鼠模型进行体内实验。然后采用免疫组织化学测定来鉴定与肿瘤组织中的EMT和WNT/β-连环蛋白信号通路相关的蛋白质。HNSCC患者肿瘤组织中P4HA3上调与不良预后呈正相关。值得注意的是,P4HA3敲低可显著抑制HNSCC的增殖和侵袭能力。通过P4HA3敲低，与EMT和WNT/β-catenin信号通路相关的基因和蛋白的水平也显著降低。重要的是,体内实验表明，P4HA3可以促进裸鼠皮下肿瘤发生，而P4HA3的敲低可以诱导肿瘤组织中EMT和WNT/β-catenin通路的显着抑制。总之,我们证明P4HA3是HNSCC的一个有前景的诊断和治疗生物标志物.作为一种致癌基因，P4HA3通过诱导EMT和激活WNT/β-catenin信号通路增加HNSCC增殖。
