Format

Send to

Choose Destination
BMC Cancer. 2018 Feb 17;18(1):193. doi: 10.1186/s12885-018-4077-3.

In vitro and in silico validation of CA3 and FHL1 downregulation in oral cancer.

Author information

1
Department of Head and Neck Surgery, A. C. Camargo Cancer Hospital, São Paulo, Brazil.
2
Laboratory of Cancer Genetics, Ludwig Institute for Cancer Research, Sao Paulo, Branch, Brazil.
3
Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos, Brazil.
4
Department of Science Biology, Universidade Federal de São Paulo, UNIFESP, Diadema, Brazil.
5
Embrapa Informatica Agropecuaria, Campinas, Brazil.
6
Discipline of Hematology and Hemotherapy, Universidade Federal de São Paulo, UNIFESP, São Paulo, Brazil.
7
Department of Head and Neck Surgery, A. C. Camargo Cancer Hospital, São Paulo, Brazil. carvalhoal@gmail.com.
8
Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos, Brazil. carvalhoal@gmail.com.
9
Department of Head and Neck Surgery, Barretos Cancer Hospital, Barretos, São Paulo, Brazil. carvalhoal@gmail.com.

Abstract

BACKGROUND:

Aberrant methylation is a frequent event in oral cancer.

METHODS:

In order to better characterize these alterations, a search for genes downregulated by aberrant methylation in oral squamous cell carcinoma (OSCC) was conducted through the mining of ORESTES dataset. Findings were further validated in OSCC cell lines and patients' samples and confirmed using TCGA data. Differentially expressed genes were identified in ORESTES libraries and validated in vitro using RT-PCR in HNSCC cell-lines and OSCC tumor samples. Further confirmation of these results was performed using mRNA expression and methylation data from The Cancer Genome Atlas (TCGA) data.

RESULTS:

From the set of genes selected for validation, CA3 and FHL1 were downregulated in 60% (12/20) and 75% (15/20) of OSCC samples, respectively, and in HNSCC cell lines. The treatment of cell lines JHU-13 and FaDu with the demethylating agent 5'-aza-dC was efficient in restoring CA3 and FHL1 expression. TCGA expression and methylation data on OSCC confirms the downregulation of these genes in OSCC samples and also suggests that expression of CA3 and FHL1 is probably regulated by methylation. The downregulation of CA3 and FHL1 observed in silico was validated in HNSCC cell lines and OSCC samples, showing the feasibility of integrating different datasets to select differentially expressed genes in silico.

CONCLUSIONS:

These results showed that the downregulation of CA3 and FHL1 data observed in the ORESTES libraries was validated in HNSCC cell lines and OSCC samples and in a large cohort of samples from the TCGA database. Moreover, it suggests that expression of CA3 and FHL1 could probably be regulated by methylation having an important role the oral carcinogenesis.

KEYWORDS:

CA3; FHL1; Gene expression; Methylation; OSCC

PMID:
29454310
PMCID:
PMC5816396
DOI:
10.1186/s12885-018-4077-3
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for BioMed Central Icon for PubMed Central
Loading ...
Support Center