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. 2021 Jun 18;11(6):908.
doi: 10.3390/biom11060908.

The m6A RNA Modification Quantity and mRNA Expression Level of RNA Methylation-Related Genes in Head and Neck Squamous Cell Carcinoma Cell Lines and Patients

Kamila Romanowska  1   2   3 , Agnieszka A Rawłuszko-Wieczorek  2 , Łukasz Marczak  4 , Agnieszka Kosińska  1 , Wiktoria M Suchorska  3 , Wojciech Golusiński  1
Affiliations

The m6A RNA Modification Quantity and mRNA Expression Level of RNA Methylation-Related Genes in Head and Neck Squamous Cell Carcinoma Cell Lines and Patients

Kamila Romanowska et al. Biomolecules. .

Abstract

RNA methylation at the nitrogen sixth of adenosine (m6A, N6-methyladenosine) is the most abundant RNA modification which plays a crucial role in all RNA metabolic aspects. Recently, m6A modification has been assigned to mediate the biological processes of cancer cells, but their significance in HNSCC development is still poorly described. Thus, the main aim of this study was to globally quantify m6A modification by the mass spectrometry approach and determine the mRNA expression level of selected m6A RNA methyltransferase (METTL3), demethylase (FTO), and m6A readers (YTHDF2, YTHDC2) in 45 HNSCC patients and 4 cell lines (FaDu, Detroit 562, A-253 and SCC-15) using qPCR. In the results, we have not observed differences in the global amount of m6A modification and the mRNA level of the selected genes between the cancerous and paired-matched histopathologically unchanged tissues from 45 HNSCC patients. However, we have found a positive correlation between selected RNA methylation machinery genes expression and m6A abundance on total RNA and characterized the transcript level of those genes in the HNSCC cell lines. Moreover, the lack of global m6A differences between cancerous and histopathologically unchanged tissues suggests that m6A alterations in specific RNA sites may specifically influence HNSCC tumorigenesis.

Keywords: RNA methylation; head and neck squamous cell carcinoma; m6A RNA modification.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
The percentage of m6A RNA modification in relation to adenosine (A) in total RNA derived from 45 head and neck squamous cancerous and paired-matched histopathologically unchanged tissues (A) and 4 cell lines (FaDu, Detroit 562, A-253, SCC-15) (B). ns (non-significant), + (mean value).
Figure 2
Figure 2
The transcript level of (A) METTL3 and (B) FTO genes in cancerous and histopathologically unchanged tissues from HNSCC patients and (C,D) cell lines. ns (non-significant), + (mean value), * (p ≤ 0.05).
Figure 3
Figure 3
Spearman correlation of m6A/A [%] and METTL3 and FTO mRNA level in cancerous tissues (A,B) and histopathologically unchanged tissues (C,D).
Figure 4
Figure 4
The transcript level of (A) YTHDF2 and (B) YTHDC2 genes in cancerous and histopathologically unchanged tissues from patients with HNSCC and cell lines (C,D). ns (non-significant), + (mean value), * (p ≤ 0.05), ** (p < 0.01).
Figure 5
Figure 5
Spearman correlation of m6A/A [%] and YTHDF2 and YTHDC2 mRNA level in cancerous tissues (A,B) and histopathologically unchanged tissues (C,D).
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