Limited evidence of human papillomavirus on breast tissue using molecular in situ methods

Abstract

BACKGROUND: Human papillomavirus (HPV) has been proposed as an etiologic agent of breast cancer based on numerous reports of high-risk (oncogenic) HPV types in malignant breast tissues. However, most of those studies used standard and nested solution polymerase chain reaction (PCR) techniques, both of which are disadvantaged by vulnerability to laboratory contamination from positive control DNA and the inability to localize the signal to a specific cell type. To overcome these drawbacks, the authors of this report explored the use of in situ molecular methods of viral detection to reassess the frequency of HPV in malignant breast tissue. METHODS: In situ hybridization (ISH) was used with probes that were specific for the capsid region of 12 oncogenic HPV types, and in situ PCR (IS-PCR) was used with primers that were specific for the capsid region of HPV-16, which is the most common oncogenic HPV type. These methods were resistant to molecular contamination and allowed identification of the positive cell type. The specimens examined were malignant tissues from patients with 70 breast cancer patients at The University of Texas M. D. Anderson Cancer Center in Houston, Texas. RESULTS: HPV was observed in 4 of 70 specimens (5.7%) using ISH and in 2 of 70 specimens (2.9%) of specimens using IS-PCR. Concordance between the 2 methods was high for negative specimens; both methods yielded negative results in 66 of 70 specimens (94.3%). However, there was no concordance for the few positive specimens, probably because of differences in sensitivity and the targeted HPV types. CONCLUSIONS: Oncogenic (high-risk) HPV types were present in malignant breast epithelium very infrequently and, thus, may be causative agents of only a relatively small proportion of all breast cancers. Cancer 2012;118:1212À20.

Is Human Papilloma Virus Associated with Breast Cancer? A Review of the Molecular Evidence

“…Other studies directly applied primers for specific oncogenic types [39, 40, 45, 46, 48, 49, 53, 62, 67, 68, 70, 72, 79]. The URR (upstream regulatory region) was also used [43], and primers for the E1 region [64, 67], microarray [56], multiplex PCR [78], hybrid capture [74], or genotyping kits directly [71].…”

The Possible Involvement of HPV in Breast Cancer

“…To overcome these disadvantages, methods of molecular biology are used, as they are resistant to contamination, like the in situ hybridization (ISH) with specific probes for the identification of the viral type in positive cell in the capsid region of the malignant mammary tissue. Based on these findings, it is obvious saying that PCR is more sensitive than the in situ hybridization or the southern blot, although hybridization in situ is more specific, as it shows the virus location (11) . (12) investigated through PCR and ISH the occurrence of HPV in breast and nipple/areola carcinoma of these patients, finding 69% and 86%, respectively, and postulate a ductular retrograde pattern of viral propagation.…”

“…Antonsson et al (2011) (22) reported the prevalence of HPV-18 DNA by PCR of 50% (27/54) (11) used PCR-IS and observed HPV-16 in 3% (2/70), and 6% by ISH (4/70), justifying the little agreement between the methods due to few positive specimens, sensibility differences, and specific HPV types.…”

“…Between 1992 and 2012 the worldwide systematic revision of a number of studies about HPV relation in breast cancer, showed that prevalence varies between 4% (3/67) in Mexico to 86% (25/29) in the USA (Table 1) (8,11,(12)(13)(14)19,21,(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36) . These variations are based in different geographic regions, and can be attributed to distinct susceptibility of the population to the various detection methods of HPV types or to the primer type of PCR used (Simões et al 2012) (37) .…”

“…Few studies have convincingly demonstrated the presence of oncogenic HPV in the human mammary epithelium using more than one method and careful methodology (11,(14)(15)(16)(17)(18)(19)(20) . Akil et al (2008) (21) investigated 113 invasive breast cancers and found 69 (61%) positive cases of high-risk HPV, as follows: HPV-16 (9%), HPV-18 (10%), HPV-31 (77%), HPV-33 (56%) and HPV-35 (37%); and 24 tissues (34,78%) amongst these specimens have been coinfected with more than one HPV type.…”