School of Basic Medicine, Jiamusi University, Jiamusi, Heilongjiang 154007, China; Tianjin Institute of Environmental and Operational Medicine, Tianjin 300050, China.
Tianjin Institute of Environmental and Operational Medicine, Tianjin 300050, China.
Pathol Res Pract. 2024 Aug;260:155481. doi: 10.1016/j.prp.2024.155481. Epub 2024 Jul 22.
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.
在这里，我们研究了脯氨酰 4-羟化酶亚基α 3（P4HA3）的作用，P4HA3 是脯氨酰-4-羟化酶（P4H）家族中最新鉴定的成员，在头颈部鳞状细胞癌（HNSCC）进展中。P4HA3 在癌症进展过程中上调；然而，其在 HNSCC 进展中的具体作用仍不清楚。因此，本研究旨在阐明 P4HA3 在 HNSCC 发生和发展中的调节功能，并描述其潜在机制。首先，我们根据癌症基因组图谱（TCGA）的微阵列数据分析了 P4HA3 的表达与 WNT 途径基因和 HNSCC 的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 中几个潜在相关的途径。然后，我们在 SCC15 和 SCC25 细胞（两种经典的 HNSCC 细胞系）中敲低 P4HA3，并使用 RT-qPCR 评估结果变化。此外，我们使用 Western blot 评估 P4HA3 在上皮间质转化（EMT）和 WNT/β-连环蛋白信号通路中的调节作用。为了探索 P4HA3 敲低对肿瘤进展的影响，我们在小鼠模型中进行了体内实验。然后使用免疫组织化学检测鉴定肿瘤组织中与 EMT 和 WNT/β-连环蛋白信号通路相关的蛋白质。HNSCC 患者肿瘤组织中上调的 P4HA3 与预后不良呈正相关。值得注意的是，P4HA3 敲低显著抑制了 HNSCC 的增殖和侵袭能力。P4HA3 敲低也显著降低了与 EMT 和 WNT/β-连环蛋白信号通路相关的基因和蛋白质的水平。重要的是，体内实验表明 P4HA3 可以促进裸鼠皮下肿瘤的发生，免疫组织化学检测显示 P4HA3 敲低显著抑制 EMT 和 WNT/β-连环蛋白通路。综上所述，我们证明 P4HA3 是 HNSCC 有前途的诊断和治疗生物标志物。作为一种癌基因，P4HA3 通过诱导 EMT 并激活 WNT/β-连环蛋白信号通路，增加 HNSCC 的增殖。
