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Human papillomavirus type 16 E7 oncoprotein mediates CCNA1 promoter methylation.

Chalertpet K, Pakdeechaidan W, Patel V, Mutirangura A, Yanatatsaneejit P - Cancer Sci. (2015)

Bottom Line: CCNA1 methylation is found in HPV-associated cervical cancers, as well as in head and neck squamous cell cancer.Using both E7 knockdown and overexpression approaches in SiHa and C33a cells, our data showed that CCNA1 promoter methylation decreases with a corresponding increase in expression in E7 siRNA-transfected cells.By contrast, CCNA1 promoter methylation was augmented with a corresponding reduction in expression in E7-overexpressing cells.

View Article: PubMed Central - PubMed

Affiliation: Human Genetics Research Group, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.

No MeSH data available.


Related in: MedlinePlus

Profile of CCNA1 promoter methylation and expression in SiHa (human papillomavirus-positive) and C33a (human papillomavirus-negative) cells. (a) Methylation status of CCNA1 promoter was detected by methylation-specific PCR using duplex primers for methylated (Met) and unmethylated (Unmet) regions. (b) CCNA1 expression was observed using duplex primers; expression of CCNA1 and GAPDH, as an internal control to normalize CCNA1 expression, were measured. Neg, distilled water as a negative control.
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fig01: Profile of CCNA1 promoter methylation and expression in SiHa (human papillomavirus-positive) and C33a (human papillomavirus-negative) cells. (a) Methylation status of CCNA1 promoter was detected by methylation-specific PCR using duplex primers for methylated (Met) and unmethylated (Unmet) regions. (b) CCNA1 expression was observed using duplex primers; expression of CCNA1 and GAPDH, as an internal control to normalize CCNA1 expression, were measured. Neg, distilled water as a negative control.

Mentions: Two cervical cancer cell lines, SiHa (HPV-positive) and C33a (HPV-negative), confirmed for their HPV status by using L1 primer (MY09/MY11),20 were subjected to CCNA1 promoter methylation and expression analysis. As shown in Figure 1a, SiHa cells showed full CCNA1 methylation, whereas only partial methylation (band intensity of 44.97%) was observed in C33a cells. For CCNA1 expression, we observed that SiHa cells had lower CCNA1 expression, giving a band intensity of 26.85%, compared with C33a (70.31%). We also used GAPDH as an internal control for normalization (Fig. 1b). The data suggest that the presence of HPV might influence CCNA1 promoter methylation and expression.


Human papillomavirus type 16 E7 oncoprotein mediates CCNA1 promoter methylation.

Chalertpet K, Pakdeechaidan W, Patel V, Mutirangura A, Yanatatsaneejit P - Cancer Sci. (2015)

Profile of CCNA1 promoter methylation and expression in SiHa (human papillomavirus-positive) and C33a (human papillomavirus-negative) cells. (a) Methylation status of CCNA1 promoter was detected by methylation-specific PCR using duplex primers for methylated (Met) and unmethylated (Unmet) regions. (b) CCNA1 expression was observed using duplex primers; expression of CCNA1 and GAPDH, as an internal control to normalize CCNA1 expression, were measured. Neg, distilled water as a negative control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4638020&req=5

fig01: Profile of CCNA1 promoter methylation and expression in SiHa (human papillomavirus-positive) and C33a (human papillomavirus-negative) cells. (a) Methylation status of CCNA1 promoter was detected by methylation-specific PCR using duplex primers for methylated (Met) and unmethylated (Unmet) regions. (b) CCNA1 expression was observed using duplex primers; expression of CCNA1 and GAPDH, as an internal control to normalize CCNA1 expression, were measured. Neg, distilled water as a negative control.
Mentions: Two cervical cancer cell lines, SiHa (HPV-positive) and C33a (HPV-negative), confirmed for their HPV status by using L1 primer (MY09/MY11),20 were subjected to CCNA1 promoter methylation and expression analysis. As shown in Figure 1a, SiHa cells showed full CCNA1 methylation, whereas only partial methylation (band intensity of 44.97%) was observed in C33a cells. For CCNA1 expression, we observed that SiHa cells had lower CCNA1 expression, giving a band intensity of 26.85%, compared with C33a (70.31%). We also used GAPDH as an internal control for normalization (Fig. 1b). The data suggest that the presence of HPV might influence CCNA1 promoter methylation and expression.

Bottom Line: CCNA1 methylation is found in HPV-associated cervical cancers, as well as in head and neck squamous cell cancer.Using both E7 knockdown and overexpression approaches in SiHa and C33a cells, our data showed that CCNA1 promoter methylation decreases with a corresponding increase in expression in E7 siRNA-transfected cells.By contrast, CCNA1 promoter methylation was augmented with a corresponding reduction in expression in E7-overexpressing cells.

View Article: PubMed Central - PubMed

Affiliation: Human Genetics Research Group, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.

No MeSH data available.


Related in: MedlinePlus