The Influence of Met Receptor Level on HGF-Induced Glycolytic Reprogramming in Head and Neck Squamous Cell Carcinoma

Abstract

Head and neck squamous cell carcinoma (HNSCC) is known to overexpress a variety of receptor tyrosine kinases, such as the HGF receptor Met. Like other malignancies, HNSCC involves a mutual interaction between the tumor cells and surrounding tissues and cells. We hypothesized that activation of HGF/Met signaling in HNSCC influences glucose metabolism and therefore substantially changes the tumor microenvironment. To determine the effect of HGF, we submitted three established HNSCC cell lines to mRNA sequencing. Dynamic changes in glucose metabolism were measured in real time by an extracellular flux analyzer. As expected, the cell lines exhibited different levels of Met and responded differently to HGF stimulation. As confirmed by mRNA sequencing, the level of Met expression was associated with the number of upregulated HGF-dependent genes. Overall, Met stimulation by HGF leads to increased glycolysis, presumably mediated by higher expression of three key enzymes of glycolysis. These effects appear to be stronger in Methigh-expressing HNSCC cells. Collectively, our data support the hypothesized role of HGF/Met signaling in metabolic reprogramming of HNSCC.

HGF-Induced PD-L1 Expression in Head and Neck Cancer: Preclinical and Clinical Findings

“…For investigating the effects of HGF stimulation in HNSCC on PD-L1 concentration, we chose three commercially available cell lines: Detroit 562, FaDu, and SCC-9. We already used these three cell lines in a project dealing with glycolytic reprogramming upon HGF stimulation [ 28 ]. As part of this project, RNA was isolated, and mRNA sequencing was performed after stimulating the cells with 50 ng/mL HGF for 16 h. Figure 1 a shows the results of this mRNA sequencing for the levels of the mRNA of CD274, the gene coding for PD-L1.…”

“…Investigations trying to determine the influence of Met on PD-L1 in other tumor diseases already found a high level of PD-L1 in Met amplified cell lines without HGF treatment [ 18 , 33 ]. In our study, we investigated the Met amplified cell line Detroit-562 [ 28 ]. It did not harbor significantly more PD-L1 mRNA or protein than the other two cell lines without HGF stimulation (see Figure 1 a,c).…”

“…After spinning down cells for 5 min at 2000 rpm and 4 °C, total RNA was isolated using a commercially available kit (RNeasy Mini Kit, Qiagen, Hilden, Germany), as described before. The detailed protocol for mRNA sequencing is described elsewhere [ 28 ]. The accession number for the raw data is GSE135552 at the Gene Expression Omnibus [ 44 ].…”

“…In the PCR reaction, 20 ng of cDNA was used with 1.5 µL of PD-L1 Primer (QuantiTect Primer Assay, Qiagen) and 12.5 µL of a ready-to-use qPCR master mix (QuantiTect SYBR Green PCR Kit, Qiagen). qPCR and analysis of results were performed as described [ 28 ].…”

Molecular phenotypes of circulating tumor cells and efficacy of nivolumab treatment in patients with head and neck squamous cell carcinoma

“…On the contrary, Wang et al suggested that c-MET expression in tumor cells suppresses the function of NK cells and T cells via the production of indoleamine-2,3-dioxygenase 18 . In addition, the activation of HGF/Met signaling in HNSCC increases glycolysis, which would result in the suppression of T-cell functions 19 . Thus, c-Met expression in tumor cells contributes to immune suppression through a variety of mechanisms.…”