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Viral load, integration and methylation of E2BS3 and 4 in human papilloma virus (HPV) 16-positive vaginal and vulvar carcinomas.

Lillsunde Larsson G, Helenius G, Sorbe B, Karlsson MG - PLoS ONE (2014)

Bottom Line: Fifty-seven samples, positive for HPV16, were selected for the study.Vaginal tumors were found to have a higher viral load (p = 0.024) compared to vulvar tumors but a high copy number (> median value, 15,000) as well as high methylation (>50%) was significantly (p = 0.010 and p = 0.045) associated with a worse cancer-specific survival rate in vulvar carcinoma, but not in vaginal carcinoma.HPV16- related parameters were found to be of clinical importance in the vulvar series only.

View Article: PubMed Central - PubMed

Affiliation: Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden; School of Health and Medical Sciences, Örebro University, Örebro, Sweden.

ABSTRACT

Objective: To investigate if viral load, integration and methylation of E2BS3 and 4 represent different ways of tumor transformation in vaginal and vulvar carcinoma and to elucidate its clinical impact.

Methods: Fifty-seven samples, positive for HPV16, were selected for the study. Detection of viral load was made with realtime-PCR using copy numbers of E6 and integration was calculated from comparing E2 to E6-copies. Methylation of E2BS3 and 4 was analysed using bisulphite treatment of tumor DNA, followed by PCR and pyrosequencing.

Results: Vaginal tumors were found to have a higher viral load (p = 0.024) compared to vulvar tumors but a high copy number (> median value, 15,000) as well as high methylation (>50%) was significantly (p = 0.010 and p = 0.045) associated with a worse cancer-specific survival rate in vulvar carcinoma, but not in vaginal carcinoma. Four groups could be defined for the complete series using a Cluster Two step analysis; (1) tumors holding episomal viral DNA, viral load below 150,000 copies not highly methylated (n = 25, 46.3%); (2) tumors harboring episomal viral DNA and being highly methylated (>50%; n = 6, 11.1%); (3) tumors with viral DNA fully integrated (n = 11, 20.4%), and (4) tumors harboring episomal viral DNA and being medium- or unmethylated (<50%) and having a high viral load (> total mean value 150,000; n = 12, 22.2%). The completely integrated tumors were found to be distinct group, whilst some overlap between the groups with high methylation and high viral load was observed.

Conclusion: HPV16- related integration, methylation in E2BS3 and 4 and viral load may represent different viral characteristics driving vaginal and vulvar carcinogenesis. HPV16- related parameters were found to be of clinical importance in the vulvar series only.

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Related in: MedlinePlus

Cancer-specific survival rate versus HPV virus copy load in vulvar carcinomas.There was a statistically significant (log-rank test; p = 0.010) difference in cancer-specific survival rate between patients with tumors showing low (< median value) and high (> median value) virus loads. Median value of viral load for vulvar series was used (15 000).
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pone-0112839-g002: Cancer-specific survival rate versus HPV virus copy load in vulvar carcinomas.There was a statistically significant (log-rank test; p = 0.010) difference in cancer-specific survival rate between patients with tumors showing low (< median value) and high (> median value) virus loads. Median value of viral load for vulvar series was used (15 000).

Mentions: To further investigate the biological differences between the cohorts, with regard to patient outcome, both series were investigated for cancer-specific survival in relation to viral load, integration and methylation of E2BS3 and 4. Despite the fact that the median number of virus copies was higher for the vaginal than for the vulvar series, a high copy number (> median value, 15 000) was highly significantly (log-rank test; p = 0.010) associated with a worse cancer-specific survival rate in vulvar carcinoma (Figure 2), but not in vaginal carcinoma (> median value 91 000, log-rank test; p = 0.551). Integration could not be significantly associated with cancer-specific survival rate in any of the two series, log-rank test; p = 0.729, p = 0.567. High methylation (>50%) was statistically significantly associated with a worse cancer-specific survival rate in vulvar carcinoma (log-rank test; p = 0.045), but not in vaginal carcinoma (log-rank test; p = 0.122).


Viral load, integration and methylation of E2BS3 and 4 in human papilloma virus (HPV) 16-positive vaginal and vulvar carcinomas.

Lillsunde Larsson G, Helenius G, Sorbe B, Karlsson MG - PLoS ONE (2014)

Cancer-specific survival rate versus HPV virus copy load in vulvar carcinomas.There was a statistically significant (log-rank test; p = 0.010) difference in cancer-specific survival rate between patients with tumors showing low (< median value) and high (> median value) virus loads. Median value of viral load for vulvar series was used (15 000).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4231157&req=5

pone-0112839-g002: Cancer-specific survival rate versus HPV virus copy load in vulvar carcinomas.There was a statistically significant (log-rank test; p = 0.010) difference in cancer-specific survival rate between patients with tumors showing low (< median value) and high (> median value) virus loads. Median value of viral load for vulvar series was used (15 000).
Mentions: To further investigate the biological differences between the cohorts, with regard to patient outcome, both series were investigated for cancer-specific survival in relation to viral load, integration and methylation of E2BS3 and 4. Despite the fact that the median number of virus copies was higher for the vaginal than for the vulvar series, a high copy number (> median value, 15 000) was highly significantly (log-rank test; p = 0.010) associated with a worse cancer-specific survival rate in vulvar carcinoma (Figure 2), but not in vaginal carcinoma (> median value 91 000, log-rank test; p = 0.551). Integration could not be significantly associated with cancer-specific survival rate in any of the two series, log-rank test; p = 0.729, p = 0.567. High methylation (>50%) was statistically significantly associated with a worse cancer-specific survival rate in vulvar carcinoma (log-rank test; p = 0.045), but not in vaginal carcinoma (log-rank test; p = 0.122).

Bottom Line: Fifty-seven samples, positive for HPV16, were selected for the study.Vaginal tumors were found to have a higher viral load (p = 0.024) compared to vulvar tumors but a high copy number (> median value, 15,000) as well as high methylation (>50%) was significantly (p = 0.010 and p = 0.045) associated with a worse cancer-specific survival rate in vulvar carcinoma, but not in vaginal carcinoma.HPV16- related parameters were found to be of clinical importance in the vulvar series only.

View Article: PubMed Central - PubMed

Affiliation: Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden; School of Health and Medical Sciences, Örebro University, Örebro, Sweden.

ABSTRACT

Objective: To investigate if viral load, integration and methylation of E2BS3 and 4 represent different ways of tumor transformation in vaginal and vulvar carcinoma and to elucidate its clinical impact.

Methods: Fifty-seven samples, positive for HPV16, were selected for the study. Detection of viral load was made with realtime-PCR using copy numbers of E6 and integration was calculated from comparing E2 to E6-copies. Methylation of E2BS3 and 4 was analysed using bisulphite treatment of tumor DNA, followed by PCR and pyrosequencing.

Results: Vaginal tumors were found to have a higher viral load (p = 0.024) compared to vulvar tumors but a high copy number (> median value, 15,000) as well as high methylation (>50%) was significantly (p = 0.010 and p = 0.045) associated with a worse cancer-specific survival rate in vulvar carcinoma, but not in vaginal carcinoma. Four groups could be defined for the complete series using a Cluster Two step analysis; (1) tumors holding episomal viral DNA, viral load below 150,000 copies not highly methylated (n = 25, 46.3%); (2) tumors harboring episomal viral DNA and being highly methylated (>50%; n = 6, 11.1%); (3) tumors with viral DNA fully integrated (n = 11, 20.4%), and (4) tumors harboring episomal viral DNA and being medium- or unmethylated (<50%) and having a high viral load (> total mean value 150,000; n = 12, 22.2%). The completely integrated tumors were found to be distinct group, whilst some overlap between the groups with high methylation and high viral load was observed.

Conclusion: HPV16- related integration, methylation in E2BS3 and 4 and viral load may represent different viral characteristics driving vaginal and vulvar carcinogenesis. HPV16- related parameters were found to be of clinical importance in the vulvar series only.

Show MeSH
Related in: MedlinePlus