Effects of 5-aza-2′deoxycytidine on RECK gene expression and tumor invasion in salivary adenoid cystic carcinoma

Abstract

Reversion-inducing cysteine-rich protein with kazal motifs (RECK), a novel tumor suppressor gene that negatively regulates matrix metalloproteinases (MMPs), is expressed in various normal human tissues but downregulated in several types of human tumors. The molecular mechanism for this downregulation and its biological significance in salivary adenoid cystic carcinoma (SACC) are unclear. In the present study, we investigated the effects of a DNA methyltransferase (DNMT) inhibitor, 5-aza-2′deoxycytidine (5-aza-dC), on the methylation status of the RECK gene and tumor invasion in SACC cell lines. Methylation-specific PCR (MSP), Western blot analysis, and quantitative real-time PCR were used to investigate the methylation status of the RECK gene and expression of RECK mRNA and protein in SACC cell lines. The invasive ability of SACC cells was examined by the Transwell migration assay. Promoter methylation was only found in the ACC-M cell line. Treatment of ACC-M cells with 5-aza-dC partially reversed the hypermethylation status of the RECK gene and significantly enhanced the expression of mRNA and protein, and 5-aza-dC significantly suppressed ACC-M cell invasive ability. Our findings showed that 5-aza-dC inhibited cancer cell invasion through the reversal of RECK gene hypermethylation, which might be a promising chemotherapy approach in SACC treatment.

Epigenetic alterations in salivary gland tumors

“…therefore, the essential function of the target drugs is blocking the function of DNA methyltransferases (Zhou et al, 2015). In a previous in vitro study utilizing ACC cell line, it was demonstrated that 5-aza-dC inhibited cancer cell invasion through the reversal of RECK (tumor suppressor gene) hypermethylation, which might be a promising chemotherapy approach in ACC treatment (Zhou et al, 2015).…”

Role of spermatogonial stem cells extract in transdifferentiation of 5-Aza-2′-deoxycytidine-treated bone marrow mesenchymal stem cells into germ-like cells

“…It seems that in application of 5‐aza‐dC more time is needed to get an acceptable response (Taranger et al, ), and the better results may be obtained by using hyper‐acetylating agents in combination with 5‐aza‐dC (Talaei‐Khozani et al, ; Talaei‐Khozani et al, ; Vojdani et al, ); furthermore, 5‐aza‐dC would be more effective on ESCs markers expression, such as OCT4, SOX2 and Nanog (Hattori et al, ; Talaei‐Khozani et al, ; Tsuji‐Takayama et al, ), and its usage in most of the studies was on cell lines, unlike our study which was on primary cell cultures (Diepeveen et al, ; Håkelien et al, ). Additionally, 5‐aza‐dC at diverse doses exerts various effects on cell morphology and gene expression (Kumar et al, ; Zhou et al, ), and also epigenetic status is varied in dissimilar animal cells (Kang et al, ), and changed with time and with environmental stimulations (Jirtle and Skinner, ; Szyf, ).…”

Aberrant methylation‐induced dysfunction of p16 is associated with osteoblast activation caused by fluoride

“…The concentrations of NaF and the time of 72 hours were chosen based on data obtained by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2.5‐iphenyltetrazolium bromide (MTT) assays. To determine if the changes of gene expression and cell cycle were affected by aberrant methylation, cells were treated with NaF together with 5‐AZA‐dC (Sigma, Missouri) at 5, 10, or 20 μmol·L −1 , for 72 hours …”