Mu Guang, Zhang Wenhao, Gu Yan, Wang Hongchang, Huang Jingjing, Bian Chengyu, Wei Ke, Xia Yang, Wang Jun
Department of Thoracic Surgery, Jiangsu Province Hospital and The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Department of Cardiothoracic Surgery, Taihe Hospital, Hubei University of Medicine, Shiyan, China.
Sci Rep. 2025 Jul 22;15(1):26672. doi: 10.1038/s41598-025-12661-9.
Prolyl 4-hydroxylase β-subunit (P4HB) is a class of molecular chaperones that inhibits the misfolding of endoplasmic reticulum proteins, and its role in microinvasive adenocarcinoma (MIA) progresses to invasive adenocarcinoma (IAC) is currently unclear. We mined the single-cell dataset GSE189357 to analyze the role of P4HB in LUAD progression. Subsequently, P4HB was overexpressed in H1299 and A549 cell lines, and Galactose lectin-3 binding protein (LGALS3BP) expression was increased and NF-kB pathway was inhibited by PCR and WB. Meanwhile, we also knock down of P4HB in H1299 and A549 cell lines, and the LGALS3BP was decreased and NF-kB pathway was activated. Afterward, the interaction between P4HB and LGALS3BP were verified by Co-IP assays. Meanwhile, the ferroptosis level and the cell proliferation, invasion and metastasis ability were detected under different P4HB expression. Finally, overexpression and knockdown P4HB stable transplants were constructed and verified in vivo. Overexpression of P4HB promoted LUAD cell proliferation (approximately two-fold) and consequently metastasis (approximately two-fold), while inhibiting ferroptosis development, as evidenced by halved ROS production. Clinically, increased P4HB is accompanied by a poor prognosis. Mechanistically, P4HB promotes LGALS3BP expression, suppressing NF-kB pathway activation and thereby driving LUAD progression. P4HB inhibited the activation of the NF-kB pathway by promoting the expression of LGALS3BP, thus promoting the progression of MIA to IAC. This provides a new target for the clinical diagnosis and treatment of LUAD.
脯氨酰4-羟化酶β亚基（P4HB）是一类分子伴侣，可抑制内质网蛋白的错误折叠，其在微浸润腺癌（MIA）进展为浸润性腺癌（IAC）中的作用目前尚不清楚。我们挖掘了单细胞数据集GSE189357，以分析P4HB在肺腺癌（LUAD）进展中的作用。随后，在H1299和A549细胞系中过表达P4HB，通过PCR和WB检测发现半乳糖凝集素-3结合蛋白（LGALS3BP）表达增加且NF-κB通路受到抑制。同时，我们也在H1299和A549细胞系中敲低P4HB，发现LGALS3BP表达降低且NF-κB通路被激活。之后，通过免疫共沉淀实验验证了P4HB与LGALS3BP之间的相互作用。同时，检测了不同P4HB表达水平下的铁死亡水平以及细胞增殖、侵袭和转移能力。最后，构建并在体内验证了P4HB过表达和敲低的稳定移植模型。P4HB的过表达促进了LUAD细胞增殖（约两倍）并因此促进了转移（约两倍），同时抑制了铁死亡的发展，活性氧生成减半证明了这一点。临床上，P4HB表达增加与预后不良相关。机制上，P4HB促进LGALS3BP表达，抑制NF-κB通路激活，从而推动LUAD进展。P4HB通过促进LGALS3BP的表达抑制NF-κB通路的激活，从而促进MIA向IAC进展。这为LUAD的临床诊断和治疗提供了新的靶点。
