Human papillomavirus-related cellular changes measured by cytometric analysis of DNA ploidy and chromatin texture.
BACKGROUND
Image cytometry has provided two highly sensitive markers for the identification of the malignant potential of squamous lesions. Aneuploidy and chromatin texture have been investigated as quantitative measures of nuclear damage in premalignant lesions and carcinoma. Real-time PCR methods have evolved to yield highly specific measurements of mRNA expression in very sparse cellular samples.


METHODS
Human papillomavirus (HPV) 16 and 18 E7 mRNA expression was measured using quantitative RT-PCR. DNA index and chromatin measures were taken from image cytology samples. The chromatin features, through discriminant analysis, were aggregated into a score, and both measurements were related to mRNA expression.


RESULTS
mRNA level and DNA index show an increasing trend over increasing histological grades. However, DNA index and chromatin score were not correlated to mRNA levels in these samples. Chromatin score differed by mRNA type found with HPV 18 infected samples having a higher score than those with HPV 16. Samples infected with HPV 16 and HPV 18 had even higher chromatin scores.


CONCLUSIONS
DNA index and chromatin score were not directly correlated with mRNA levels. However both mRNA and DNA index were related to histological grade, and chromatin score was associated with HPV type. Therefore, DNA index and mRNA levels could be independent predictors of cervical dysplasia, and chromatin score could be related to the viral integration process in cells infected with HPV 18 or dual infections.
Scheurer
Michael E
ME
Department of Epidemiology, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA.
Guillaud
Martial
M
Tortolero-Luna
Guillermo
G
McAulay
Calum
C
Follen
Michele
M
Adler-Storthz
Karen
K
D000328:Adult; D002277:Carcinoma / Q000235:genetics* / Q000473:pathology / Q000821:virology; D002843:Chromatin / Q000235:genetics / Q000473:pathology*; D002869:Chromosome Aberrations; D005260:Female; D006367:HeLa Cells; D006801:Humans; D019044:Image Cytometry / Q000379:methods; D008875:Middle Aged; D030361:Papillomavirus Infections / Q000235:genetics*; D011003:Ploidies; D011237:Predictive Value of Tests; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D002583:Uterine Cervical Neoplasms / Q000235:genetics* / Q000473:pathology / Q000821:virology; D016662:Virus Integration / Q000235:genetics
D002843:Chromatin; D012333:RNA, Messenger; D012367:RNA, Viral