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Methylation-associated gene silencing of RARB in areca carcinogens induced mouse oral squamous cell carcinoma.

Lai ZL, Tsou YA, Fan SR, Tsai MH, Chen HL, Chang NW, Cheng JC, Chen CM - Biomed Res Int (2014)

Bottom Line: These results showed that retinoic acid receptor β (RARB) was indicated in hypermethylation at the promoter region and the loss of expression during cancer development.According to the results of real-time PCR, it was shown that de novo DNA methyltransferases were involved in gene epigenetic alternations of OSCC.Collectively, our results showed that RARB hypermethylation was involved in the areca-associated oral carcinogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Sciences, Agricultural Biotechnology Center, National Chung Hsing University, No. 250 Kao-Kuang Road, Taichung 402, Taiwan.

ABSTRACT
Regarding oral squamous cell carcinoma (OSCC) development, chewing areca is known to be a strong risk factor in many Asian cultures. Therefore, we established an OSCC induced mouse model by 4-nitroquinoline-1-oxide (4-NQO), or arecoline, or both treatments, respectively. These are the main two components of the areca nut that could increase the occurrence of OSCC. We examined the effects with the noncommercial MCGI (mouse CpG islands) microarray for genome-wide screening the DNA methylation aberrant in induced OSCC mice. The microarray results showed 34 hypermethylated genes in 4-NQO plus arecoline induced OSCC mice tongue tissues. The examinations also used methylation-specific polymerase chain reaction (MS-PCR) and bisulfite sequencing to realize the methylation pattern in collected mouse tongue tissues and human OSCC cell lines of different grades, respectively. These results showed that retinoic acid receptor β (RARB) was indicated in hypermethylation at the promoter region and the loss of expression during cancer development. According to the results of real-time PCR, it was shown that de novo DNA methyltransferases were involved in gene epigenetic alternations of OSCC. Collectively, our results showed that RARB hypermethylation was involved in the areca-associated oral carcinogenesis.

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DNMTs RNA expression in human oral cancer cell lines. The measurement of DNMTs expression levels by real-time PCR was showed in (a), (b), and (c), respectively. (a) The Dnmt1 expression was not changed in all human oral cancer cell lines. (b) and (c) TW2.6 and Ca922 were higher expression in Dnmt3a and Dnmt3b than others. The figures shown are the mean of three experiments where all of the samples were analyzed in triplicate. The start sign showed the statistically significant (P < 0.05).
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fig5: DNMTs RNA expression in human oral cancer cell lines. The measurement of DNMTs expression levels by real-time PCR was showed in (a), (b), and (c), respectively. (a) The Dnmt1 expression was not changed in all human oral cancer cell lines. (b) and (c) TW2.6 and Ca922 were higher expression in Dnmt3a and Dnmt3b than others. The figures shown are the mean of three experiments where all of the samples were analyzed in triplicate. The start sign showed the statistically significant (P < 0.05).

Mentions: DNA methylation is catalyzed by the family of DNA methyltransferases (DNMT) including Dnmt1, Dnmt3a, and Dnmt3b. Figure 5 shows the measurement of gene expressions via real-time PCR on Dnmt1, Dnmt3a, and Dnmt3b in human oral cancer lines. For Dnmt1, there were no expression differences between NHOK and other cell lines (Figure 5(a)). However, Dnmt3a and Dnmt3b showed significantly higher expression levels in TW2.6 and Ca922 than others (Figures 5(b) and 5(c)).


Methylation-associated gene silencing of RARB in areca carcinogens induced mouse oral squamous cell carcinoma.

Lai ZL, Tsou YA, Fan SR, Tsai MH, Chen HL, Chang NW, Cheng JC, Chen CM - Biomed Res Int (2014)

DNMTs RNA expression in human oral cancer cell lines. The measurement of DNMTs expression levels by real-time PCR was showed in (a), (b), and (c), respectively. (a) The Dnmt1 expression was not changed in all human oral cancer cell lines. (b) and (c) TW2.6 and Ca922 were higher expression in Dnmt3a and Dnmt3b than others. The figures shown are the mean of three experiments where all of the samples were analyzed in triplicate. The start sign showed the statistically significant (P < 0.05).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150525&req=5

fig5: DNMTs RNA expression in human oral cancer cell lines. The measurement of DNMTs expression levels by real-time PCR was showed in (a), (b), and (c), respectively. (a) The Dnmt1 expression was not changed in all human oral cancer cell lines. (b) and (c) TW2.6 and Ca922 were higher expression in Dnmt3a and Dnmt3b than others. The figures shown are the mean of three experiments where all of the samples were analyzed in triplicate. The start sign showed the statistically significant (P < 0.05).
Mentions: DNA methylation is catalyzed by the family of DNA methyltransferases (DNMT) including Dnmt1, Dnmt3a, and Dnmt3b. Figure 5 shows the measurement of gene expressions via real-time PCR on Dnmt1, Dnmt3a, and Dnmt3b in human oral cancer lines. For Dnmt1, there were no expression differences between NHOK and other cell lines (Figure 5(a)). However, Dnmt3a and Dnmt3b showed significantly higher expression levels in TW2.6 and Ca922 than others (Figures 5(b) and 5(c)).

Bottom Line: These results showed that retinoic acid receptor β (RARB) was indicated in hypermethylation at the promoter region and the loss of expression during cancer development.According to the results of real-time PCR, it was shown that de novo DNA methyltransferases were involved in gene epigenetic alternations of OSCC.Collectively, our results showed that RARB hypermethylation was involved in the areca-associated oral carcinogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Sciences, Agricultural Biotechnology Center, National Chung Hsing University, No. 250 Kao-Kuang Road, Taichung 402, Taiwan.

ABSTRACT
Regarding oral squamous cell carcinoma (OSCC) development, chewing areca is known to be a strong risk factor in many Asian cultures. Therefore, we established an OSCC induced mouse model by 4-nitroquinoline-1-oxide (4-NQO), or arecoline, or both treatments, respectively. These are the main two components of the areca nut that could increase the occurrence of OSCC. We examined the effects with the noncommercial MCGI (mouse CpG islands) microarray for genome-wide screening the DNA methylation aberrant in induced OSCC mice. The microarray results showed 34 hypermethylated genes in 4-NQO plus arecoline induced OSCC mice tongue tissues. The examinations also used methylation-specific polymerase chain reaction (MS-PCR) and bisulfite sequencing to realize the methylation pattern in collected mouse tongue tissues and human OSCC cell lines of different grades, respectively. These results showed that retinoic acid receptor β (RARB) was indicated in hypermethylation at the promoter region and the loss of expression during cancer development. According to the results of real-time PCR, it was shown that de novo DNA methyltransferases were involved in gene epigenetic alternations of OSCC. Collectively, our results showed that RARB hypermethylation was involved in the areca-associated oral carcinogenesis.

Show MeSH
Related in: MedlinePlus