Endoplasmic reticulum stressed HNSCC cell-derived exosomal miR-26a-5p promotes PD-L1 expression in macrophage through PTEN/AKT signaling pathway
JIAO Pengfei,
WANG Zeyu,
WU Heming,
YAO Siyue,
WANG Huilin,
YAO Enhui,
ZHANG Yuyao,
YUAN Yi,
ZHONG Yi
Affiliations
JIAO Pengfei
1.The Affiliated Stomatology Hospital of Suzhou Vocational Health College.2.Jiangsu Province Key Laboratory of Oral Diseases, School of Stomatology, Nanjing Medical University 3.Jiangsu Province Engineering Research Center of Stomatological Translational Medicine, School of Stomatology, Nanjing Medical University
WANG Zeyu
1Jiangsu Province Key Laboratory of Oral Diseases, School of Stomatology, Nanjing Medical University. 2.Jiangsu Province Engineering Research Center of Stomatological Translational Medicine, School of Stomatology, Nanjing Medical University
WU Heming
1.Jiangsu Province Key Laboratory of Oral Diseases, School of Stomatology, Nanjing Medical University 2.Jiangsu Province Engineering Research Center of Stomatological Translational Medicine, School of Stomatology, Nanjing Medical University 3.Department of Oral and Maxillofacial Surgery, the Affiliated Stomatological Hospital of Nanjing Medical University
YAO Siyue
1.The Affiliated Stomatology Hospital of Suzhou Vocational Health College 2.Jiangsu Province Key Laboratory of Oral Diseases, School of Stomatology, Nanjing Medical University 3.Jiangsu Province Engineering Research Center of Stomatological Translational Medicine, School of Stomatology, Nanjing Medical University
WANG Huilin
1.Jiangsu Province Key Laboratory of Oral Diseases, School of Stomatology, Nanjing Medical University 2.Jiangsu Province Engineering Research Center of Stomatological Translational Medicine, School of Stomatology, Nanjing Medical University
YAO Enhui
1.Jiangsu Province Key Laboratory of Oral Diseases, School of Stomatology, Nanjing Medical University. 2.1
ZHANG Yuyao
1.Jiangsu Province Key Laboratory of Oral Diseases, School of Stomatology, Nanjing Medical University 2.1
YUAN Yi
1.Jiangsu Province Key Laboratory of Oral Diseases, School of Stomatology, Nanjing Medical University 2.Jiangsu Province Engineering Research Center of Stomatological Translational Medicine, School of Stomatology, Nanjing Medical University 3.Department of Oral and Maxillofacial Surgery, the Affiliated Stomatological Hospital of Nanjing Medical University
ZHONG Yi
of Stomatology, Nanjing Medical University 2.Jiangsu Province Engineering Research Center of Stomatological Translational Medicine, School of Stomatology, Nanjing Medical University 3.Department of Oral Pathology, the Affiliated Stomatological Hospital of Nanjing Medical University
Objective To investigate the impact of exosomal miRNAs derived from endoplasmic reticulum-stressed (ERS) head and neck squamous cell carcinoma (HNSCC) cells on macrophages. Methods This study was reviewed and approved by the Ethics Committee. The expression levels of ERS-associated proteins, including protein kinase R-like endoplasmic reticulum kinase (PERK) and glucose-regulated protein 78 (GRP78), in HNSCC tissues and para-tumor tissues were detected by Western blot (WB) and quantitative real-time PCR (RT-qPCR). HN4 human laryngeal squamous cell carcinoma cells were treated with 500 U/mL interferon-γ (IFN-γ) for 48 h to induce ER stress, and exosomes secreted by ER-stressed HN4 cells were collected and identified. The types of miRNAs in exosomes were identified through bioinformatics analysis, and the target genes of miRNAs were predicted. Macrophages were transfected with miRNA, co cultured with collected exosomes, and the expression of PTEN in macrophages was knocked down. The downstream signaling pathway regulated by exosomal miRNAs was studied by WB and RT-qPCR. Results Compared with that in para-tumor tissues, the expression level of ER stress-associated proteins in HNSCC tissues was increased (P<0.05). RNA-seq analysis revealed that miR-26a-5p was highly upregulated in ER-stressed HN4 cell-derived exosomes (P<0.05). PTEN is the target gene for miR-26a-5p. miR-26a-5p increased the expression level of PD-L1 in macrophages and downregulated the expression of PTEN (P<0.05). Macrophages co cultured with ERS extracellular vesicles showed an increase in miR-26a-5p and PD-L1 expression, a decrease in PTEN expression, and an increase in p-AKT expression (P<0.05). Knock down the expression of PTEN in macrophages and increase the expression of PD-L1 (P<0.01). Conclusion ERS HNSCC cell-derived exosomal miR-26a-5p promotes the expression of PD-L1 in macrophages through the PTEN/AKT signaling pathway.
