Why are papilloma viruses associated with cancer

Four-micron thick sections of breast tumour tissue were cut and placed on silanised slides. Positive controls known virus-positive cervical cancer tissue , and negative controls known virus-negative tissue , and controls omitting DIGdUTP to confirm incorporation of DIG label , omitting primers to confirm the signal was the result of specific amplification rather than self priming of degraded tissues , or omitting Taq polymerase, were also undertaken for each specimen.

The outcomes were assessed by the staining of the DIG label in the tissues by light microscopy. A number of specimens gave a false-positive signal when the primers were omitted. Consequently, we carried out a no-primer negative control for each sample at the same time as the in situ screening PCR and eliminated the specimens that gave false-positive colour signals. Any specimens that produced a signal in the in situ PCR negative controls omission of the DNA primers and separately, omission of the Taq polymerase were eliminated from the study.

False positives indicated that the DNA was self-priming and were unsuitable for in situ work this is probably because of fragmented DNA, caused by formalin fixation, acting as primers. The methods used are outlined below. The products were sequenced and the identity of the sequences was determined using the BLAST alignment system. Previously described protocols were used to extract genomic DNA from the breast cancer specimens Dawkins et al , The amplified products were visualised on a 1.

The PCR was independently repeated for each sample. The presence of koilocytosis in the fixed breast cancer series was assessed by light microscopy with koilocyte positive cervical cancer specimens used for comparison. Koilocytes were best characterised by Koss and Durfee, the presence of large cells with relatively small, but irregular and hyperchromatic nuclei surrounded by clear and transparent cytoplasm.

Koilocytosis is restricted to the replicating basal cells and multinucleation is common in these cells. The histological features of HPV positive breast tumours are similar to cervical koilocytosis and have previously been reported by de Villiers et al Polymerase chain reaction PCR product nucleotide sequences of human papilloma virus HPV -positive patient samples and breast cancer cell lines.

There is no sequence variation in specimen 7 when matched against HPV-type 16 genome accession FJ Lanes 1—7 are patient samples breast cancer specimens 1—7. Lanes 9—11 are negative controls water in place of DNA in reaction.

Both samples in lanes 1 specimen 1 and 3 specimen 3 are negative for HPV. Having established the method and demonstrated the presence of HPV in breast cancer cell lines, we screened a series of fixed breast cancer and normal breast tissue specimens using in situ PCR. As discussed in Materials and Methods section, we eliminated those samples, which potentially could give false-positive HPV results, identified by positive in situ PCR without primers.

An unknown number of these eliminated specimens would have been true positives. Accordingly, the data cannot be used to make estimates of prevalence of the presence of these viruses. Unexpectedly, we saw HPV containing cells in the surrounding normal tissue of some samples Figure 4D.

The presence of HPV in normal breast tissues is consistent with the requirement for HPV infection in the breast tissue before HPV-induced tumourigenic transformation of a single clone. Human papilloma virus HPV in cancer cells of ductal carcinoma in situ breast cancer demonstrated by in situ polymerase chain reaction PCR same specimen in all panels. The appearance of koilocytes in the HPV containing cells shown in panel F selected koilocytes shown by arrows is indicated by the clearing of the cytoplasm and condensed, hyperchromatic nuclei.

Seven breast cancer specimens and three normal specimens showed false-positive outcomes when the PCR primer was omitted from the in situ PCR analysis. These specimens were not considered further. The outcome was confirmed in all the three HPV-positive normal breast specimens. The outcome was confirmed in five of the eight HPV-positive breast cancer specimens. Minor sequence variations indicate that contamination was unlikely. The identification of two HPV types is a further indication that contamination is unlikely.

All controls gave the expected outcomes. Koilocyte-like cells were also observed in some HPV-positive normal breast tissue specimens. How is it transmitted? How can it be recognised? What are the symptoms? What consequences can it cause and why is it worth getting vaccinated? Papilloma virus: what are HPVs? How is HPV transmitted?

In the absence of a proper screening test for the infection as opposed to a Pap test or HPV test for women , examinations such as: anoscopy; anal swab; the acetic acid test; peniscopy; molecular biology investigations for HPV viral DNA. The link between HPV and male urogenital cancers There is some evidence to suggest a link between the presence of the Papilloma Virus and the development of urogenital cancers in men, including penile cancer, testicular cancer and urothelial carcinomas.

Papilloma virus and penile carcinoma Penile cancer is a relatively rare disease in Western countries, with an incidence of between 0. Study Findings. Metastatic Cancer Research.

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