P2Y2 receptors mediate nucleotide-induced EGFR phosphorylation and stimulate proliferation and tumorigenesis of head and neck squamous cell carcinoma cell lines. (October 2020)

• Record Type: Journal Article
• Authors: Woods, Lucas T.; Jasmer, Kimberly J.; Muñoz Forti, Kevin; Shanbhag, Vinit C.; Camden, Jean M.; Erb, Laurie; Petris, Michael J.; Weisman, Gary A.
• Is Part Of: Oral oncology. Volume 109(2020)
• Publication Date: 2020-10
• Subjects: Head and Neck Squamous Cell Carcinoma -- Purinergic P2Y2 Receptors -- Purinergic P2Y2 Receptor Antagonist -- Epidermal Growth Factor Receptor -- Adenosine 5′-Triphosphate -- Tumor Microenvironment -- Cetuximab; Mouth -- Cancer -- Periodicals; Mouth -- Tumors -- Periodicals; Mouth Diseases -- Periodicals; Mouth Neoplasms -- Periodicals; Bouche -- Cancer -- Périodiques; Bouche -- Tumeurs -- Périodiques; Tumeurs -- Périodiques; Electronic journals; 616.9943105
• DOI: 10.1016/j.oraloncology.2020.104808
• Languages: English
• ISSNs: 1368-8375

Abstract

Highlights: P2Y2 Rs mediate EGFR and ERK1/2 phosphorylation in CAL27 and FaDu HNSCC cells. Metalloprotease or Src inhibition blocks nucleotide-induced EGFR phosphorylation. P2Y2 R knockout attenuates CAL27 and FaDu tumor xenograft growth. Pharmacological P2Y2 R inhibition reduces tumorigenesis of mouse oral cancer cells. P2Y2 Rs represent a novel therapeutic target to treat HNSCC. Abstract: Objectives: To assess functional expression of the P2Y2 nucleotide receptor (P2Y2 R) in head and neck squamous cell carcinoma (HNSCC) cell lines and define its role in nucleotide-induced epidermal growth factor receptor (EGFR) transactivation. The use of anti-EGFR therapeutics to treat HNSCC is hindered by intrinsic and acquired drug resistance. Defining novel pathways that modulate EGFR signaling could identify additional targets to treat HNSCC. Materials and Methods: In human HNSCC cell lines CAL27 and FaDu and the mouse oral cancer cell line MOC2, P2Y2 R contributions to extracellular nucleotide-induced changes in intracellular free Ca 2+ concentration and EGFR and extracellular signal-regulated kinase (ERK1/2) phosphorylation were determined using the ratiometric Ca 2+ indicator fura-2 and immunoblot analysis, respectively. Genetic knockout of P2Y2 Rs using CRISPR technology or pharmacological inhibition with P2Y2 R-selective antagonist AR-C118925 defined P2Y2 R contributions to in vivo tumor growth. Results: P2Y2 R agonists UTP and ATP increased intracellular Ca 2+ levels and ERK1/2 and EGFR phosphorylation in CAL27 and FaDu cells, responses that were inhibited by AR-C118925 or P2Y2 R knockout. P2Y2 R-mediated EGFR phosphorylation was also attenuated by inhibition of the adamalysin family of metalloproteases or Src family kinases. P2Y2 R knockout reduced UTP-induced CAL27 cell proliferation in vitro and significantly reduced CAL27 and FaDu tumor xenograft volume in vivo . In a syngeneic mouse model of oral cancer, AR-C118925 administration reduced MOC2 tumor volume. Conclusion: P2Y2 Rs mediate HNSCC cell responses to extracellular nucleotides and genetic or pharmacological blockade of P2Y2 R signaling attenuates tumor cell proliferation and tumorigenesis, suggesting that the P2Y2 R represents a novel therapeutic target in HNSCC.