Human Papillomavirus Infection, Centrosome Aberration, and Genetic Stability in Cervical Lesions

Abstract

DNA replication and centrosome duplication have to be strictly synchronized to guarantee genomic stability. p53, pRb, cyclin E, and cyclin A are reported to be involved in the synchronizing process. We investigated the relationship between papillomavirus infection, centrosome aberration and aneuploidy during genesis of cervical carcinoma. The number of centrosomes found in cells from normal cervical epithelium (n ‫؍‬ 5), condyloma acuminata (n ‫؍‬ 5), cervical intraepithelial neoplasia (CIN) I, II, and III (n ‫؍‬ 14) and invasive cervical carcinoma (n ‫؍‬ 5) was analyzed by ␥ tubulin immunofluorescence staining. The nuclear DNA content was investigated by image cytometry and human papillomavirus (HPV) infection was determined by polymerase chain reaction. Normal epithelia and condyloma acuminata showed cells with one or two centrosomes, whereas CIN lesions showed cells with an increasing number of centrosomes. This abnormality was found to be lowest in CIN I lesions, increased with advancing grade of CIN and was highest in lesions of invasive carcinomas. In parallel, an increasing number of cells with aberrant DNA content was seen. All carcinomas and all except one of the CIN III lesions showed aneuploidy. Three CIN II cases were aneuploid and two cases with CIN I were tetraploid. Normal epithelia and condyloma acuminata showed diploidy. All invasive carcinomas and lesions with CIN were positive for high-risk HPV types 16, 18, or 31, except one invasive carcinoma and one CIN II lesion positive for universal primers only. Three condyloma acuminata were HPV 16-positive and one HPV 6-positive.The results suggest that high-risk HPV infection is correlated to a progressive numerical disturbance of centrosome replication followed by progressive chromosomal aberrations in CIN lesions and invasive carcinomas.

Characterisation of breast fine-needle aspiration biopsies by centrosome aberrations and genomic instability

“…We found that scattering of DNA profiles as a measure of genomic instability correlates highly significantly with centrosome aberrations. While centrosome abnormalities have been reported earlier in different human malignancies (Pihan et al, 1998;Ghadimi et al, 2000;Skyldberg et al, 2001;Roshani et al, 2002), including breast cancer (Lingle et al, 1998;Lingle and Salisbury, 1999), a difference in the extent of centrosome aberrations in cytometrically determined subtypes of breast carcinomas has, to our knowledge, not been reported by other authors.…”

Centrosome amplification and the development of cancer

“…The degree of genomic instability parallels the degree of centrosome abnormalities in cell lines from breast , pancreas , prostate (Pihan et al, 2001), colon (Ghadimi et al, 2000), and cervix tumors (Skyldberg et al, 2001), from short-term culture of mouse mammary tumors (Montagna et al, 2002), and from SV40 ST overexpressing fibroblasts (Gaillard et al, 2001). Centrosome abnormalities were higher in high-grade prostate tumors (Pihan et al, 2001) and high-grade cervical tumors (Skyldberg et al, 2001) than in low-grade tumors. Centrosome amplification and chromosomal instability can be induced by over-expression of the centrosomal protein pericentrin and by over-expression of the centrosome kinase BTAK/STK15, suggesting that alternative mechanisms may lead to centrosome defects and consequent genomic instability (Pihan et al, 2001;Zhou et al, 1998).…”

“…Strong support for a direct mechanistic link between centrosome amplification and CIN is suggested by the significant linear correlation between centrosome amplification and the rate of change in karyotype (CIN) seen in human breast tumors . Although such correlation alone does not necessarily imply cause and effect, these observations have led many authors to propose the hypothesis that centrosome amplification is the primary cause of genomic instability observed in most tumors (Gaillard et al, 2001;Lingle et al, 2002;Sato et al, 2001;Skyldberg et al, 2001). An alternative hypothesis has been proposed that chromosomal instability seen in cancer cells is caused by aneuploidy, that is that aneuploidy itself destabilizes the karyotype and thus initiates CIN leading to widespread heterogeneity in tumor cell phenotypes (Duesberg, 1999;Duesberg and Rasnick, 2000;Duesberg et al, 1998;Li et al, 2000).…”

Human papillomaviruses and centrosome duplication errors: modeling the origins of genomic instability

“…It has been shown that these multipolar mitoses are associated with abnormal numbers of mitotic spindle poles that are formed by centrosomes (Duensing et al, 2000). The incidence of cells with abnormal centrosome numbers increases in parallel with clinical progression during HPV-associated cervical carcinogenesis (Skyldberg et al, 2001).…”