Srisuttee, Ratakorn

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Srisuttee, Ratakorn
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ratakorn.sr@kmitl.ac.th

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    Head and neck squamous cell carcinoma drives long interspersed element-1 hypomethylation in the peripheral blood mononuclear cells
    (2019-01-01)
    Arayataweegool, Areeya
    ;
    Srisuttee, Ratakorn
    ;
    Mahattanasakul, Patnarin
    ;
    Tangjaturonsasme, Napadon
    ;
    Kerekhanjanarong, Virachai
    Objective: Alteration of long interspersed element-1 (LINE-1) methylation in peripheral blood mononuclear cells (PBMCs) has been simultaneously activated to breast carcinogenesis due to its secretion. To evaluate the effect in head and neck squamous cell carcinoma (HNSCC), LINE-1 methylation levels and patterns have been measured both in vitro and in vivo. Methods: Analysis of LINE-1 methylation in cocultured models between HNSCC cell lines and normal PBMCs was performed. The observation of PBMCs of HNSCC patients compared to PBMCs of normal controls was performed using the semiquantitative combined bisulfite restriction analysis technique. Results: Downregulation of LINE-1 methylation was significantly found in the PBMCs cocultured with all HNSCC cell lines compared to normal PBMCs. Likewise, a reduction in LINE-1 methylation levels was observed in PBMCs of HNSCC compared to normal PBMCs (p < 0.0001). Receiver operating characteristic analysis demonstrated the potential of the unmethylated alleles (<sup>u</sup>C<sup>u</sup>C) of LINE-1 for distinguishing the PBMCs of HNSCC patients from normal controls with 100% sensitivity and specificity. Conclusion: Our data supported that the alteration of LINE-1 methylation levels in PBMCs was influenced by HNSCC secretions. Moreover, the unmethylated LINE-1 allele of PBMCs was proved to be an effective tumor marker and possesses a potential as HNSCC diagnostic tool.
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    Evaluation of NID2 promoter methylation for screening of Oral squamous cell carcinoma
    (2020-03-14)
    Srisuttee, Ratakorn
    ;
    Arayataweegool, Areeya
    ;
    Mahattanasakul, Patnarin
    ;
    Tangjaturonrasme, Napadon
    ;
    Kerekhanjanarong, Virachai
    Background: Oral squamous cell carcinoma (OSCC) is an aggressive human malignancy. Because of late diagnosis and recurrence of OSCC, the treatment of patients with OSCC is often ineffective. Thus, finding novel biomarkers of OSCC are essential. Here we derived a methylation marker by utilizing methylation microarray data and testing its capacity in cross-sectional study designed for OSCC detection and screening. Methods: According to bioinformatics analysis of total of 27,578 cg sites, cg22881914 of Nidogen 2 (NID2) methylation was selected for evaluation. Next, we confirmed the methylation status by bisulfite sequencing from the microdissected OSCC cells in comparison with the microdissected oral epithelia. Subsequently, we developed a simple technique using real-time PCR with the specific probe to examine the ability for the detection of OSCC in the oral epithelial samples, which included 103 oral rinse and 82 oral swab samples. Results: Based on the comparison of microdissected tissue, cg22881914 of NID2 was proved to be methylated in most OSCC cells but unmethylated in the normal oral epithelia. Furthermore, the methylated NID2-relied quantitative PCR approach has demonstrated that this marker assists in distinguishing among patients with OSCC from normal oral epithelia, smokers, and patients with oral lichen planus using the non-invasive oral rinse and swab samples. Conclusions: Specific methylation at cg22881914 of NID2 of OSCC could be used as an important potential marker for detecting OSCC. Thus, to certify the utility of this marker, further studies with a larger sample size are needed.
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    Induction of ZCCHC6 expression in peripheral blood mononuclear cells by HNSCC secretions
    (2020-09-05)
    Arayataweegool, Areeya
    ;
    Srisuttee, Ratakorn
    ;
    Bin-Alee, Fardeela
    ;
    Mahattanasakul, Patnarin
    ;
    Tangjaturonrasme, Napadon
    Cancer secretion can change the properties of adjacent cells, including peripheral blood mononuclear cells (PBMCs). We investigated whether such secretion influences messenger RNA expression in PBMCs of patients with head and neck squamous cell carcinoma (HNSCC). In the present study, co-culture model of normal PBMCs and HNSCC cell lines were established. The PBMCs were subsequently subjected to RNA sequencing for transcriptome analysis. Furthermore, expression data from the Gene Expression Omnibus repository, platform GPL4133, series GSE39400, were gathered to analyze, afterward identify zinc finger CysCysHisCys (CCHC)–type domain-containing protein 6 (ZCCHC6) as the main gene involved in HNSCC. This gene was then validated by a quantitative real-time polymerase chain reaction. The results showed that ZCCHC6 was expressed at significantly higher levels in the patients with HNSCC than in the healthy controls, and the sensitivity and specificity of these findings for diagnostic purposes were 100.00% and 70.83%, respectively. In summary, our findings demonstrated that the secretion of HNSCC cells could cause the alterations in messenger RNA expression by PBMCs. The ZCCHC6 expression level may apply in HNSCC screening.