Cervical carcinomas with a micropapillary component: a clinicopathological study of eight cases.

AIMS: Micropapillary carcinomas, or carcinomas with a micropapillary component, are well recognised in the breast and other anatomical sites. However, they have seldom been described in the cervix. In this article, we present a clinicopathological analysis of eight cervical tumours that showed at least a focal (≥5%) component of micropapillary carcinoma. METHODS AND RESULTS: The study group comprised eight cervical carcinomas (four adenocarcinomas and four adenosquamous carcinomas) with a micropapillary component. The median patient age was 41.5 years (range 27-65 years). At presentation, five patients were stage IB, two were stage IIB, and one was stage IV. The micropapillary component accounted for ≤25% of the tumour on initial biopsy or resection specimens in all but one case. Immunohistochemistry showed 'inside-out' (reverse polarity) mucin 1 staining along the cell membrane abutting the stroma. Four patients developed metastasis, all of whom showed a pure micropapillary pattern; this led to a misdiagnosis of an apparently independent peritoneal serous carcinoma in one case. All tumours showed diffuse p16 expression, and all three cases that were tested were positive for human papillomavirus (HPV) 18. Three of the six patients with at least 12 months of follow-up died of disease, and one is alive with distant metastasis. CONCLUSIONS: Usual-type (HPV-related) cervical carcinomas may show micropapillary differentiation, usually as a focal finding, and the cells show reverse polarity like similar tumours arising in other sites. Micropapillary cervical carcinoma appears to be a clinically aggressive malignancy, although this needs to be confirmed in larger studies.

Infection and persistence of erythrovirus B19 in benign and cancerous thyroid tissues.

Human erythrovirus B19 (EVB19) is a small, pathogenic DNA virus that has been associated with a wide range of illnesses. The primary site of replication is in bone marrow-derived erythroid progenitor cells, but EVB19 DNA has been detected in a wide range of organs. Recently, studies have linked EVB19 to thyroid cancers and other thyroid diseases. Previous studies from multiple laboratories have detected EVB19 capsid proteins in Graves' disease, Hashimoto's thyroiditis, and thyroid cancer tissues. Data on viral gene expression and mechanism of infection in the thyroid are lacking. To investigate EVB19 infection and persistence in the thyroid, previously archived adult and pediatric tissue sections were examined for EVB19 DNA, RNA, and capsid proteins, as well as EVB19 receptor P-antigen and co-receptor α5ß1 integrin. EVB19 DNA and protein were detected in a majority of tissues examined (87% and 68%, respectively). Detection was similar in adult and pediatric samples. Quantification of viral genomes revealed no significant difference in the amount of viral DNA in benign, cancerous, or metastatic thyroid tissues. EVB19 capsid RNA was detected in 67% of the tissues examined, confirming at least low-level viral gene expression. Immunohistochemical staining for P-antigen and α5ß1 detected the receptor and co-receptor most frequently on normal thyroid epithelial cells. EVB19 capsid staining could be detected in tumors lacking viral receptors. These results suggest that normal thyroid epithelial cells are the initial target for EVB19 infection in the thyroid and allow for continued persistence in both normal and cancerous thyroid tissues.

No evidence of association between human cytomegalovirus infection and papillary thyroid cancer.

BACKGROUND: Human cytomegalovirus (CMV) has been detected in the thyroid gland and thyroid tumors. CMV infection may activate the mitogen-activated protein kinase pathway, of which aberrant activation is frequently associated with BRAF mutation in papillary thyroid cancer. METHODS: A total of 45 paired tumorous and adjacent non-neoplastic tissue samples, including 5 follicular adenoma and 40 papillary thyroid cancer, were obtained during thyroidectomy. BRAF mutational status was determined using direct sequencing. The presence of CMV DNA was determined using conventional PCR and quantitative real-time PCR. CMV protein in the tissue samples were evaluated with Western blot analysis. RESULTS: BRAF mutation was identified in the cancerous part of 31 (78%) papillary thyroid cancers. Papillary cancer with BRAF mutation was significantly associated with a larger tumor size (P = 0.045), extrathyroidal invasion (P = 0.012), lymph node metastasis (P = 0.008), and a higher TNM stage (P = 0.044). CMV DNA and protein were not detected in any studied samples. CONCLUSIONS: Our results suggest no association between CMV infection and papillary thyroid cancer.

Demonstration of Epstein-Barr Virus by In Situ Hybridization in Papillary Thyroid Carcinomas Developing on Background of Hashimoto's Thyroiditis.

AIM: This study aimed to demonstrate the role of Epstein-Barr Virus (EBV) in papillary thyroid carcinomas (PTC) developing on the background of Hashimoto's thyroiditis (HT). METHODS: The presence of EBV in tumoral tissue, lymphocytes, and peritumoral normal thyroid tissue was investigated using the in situ hybridization method in paraffin blocks. The subtypes of PTC, tumor diameter, TNM stage, multifocality, invasion of thyroid capsule, perineural invasion, and muscular tissue invasion were identified and compared according to EBV involvement. RESULTS: Eighty-one patients with HT diagnosis, with 93.8% (n=76) female and 6.2% (n=5) male, were included in the study. Papillary microcarcinoma was the pathological diagnosis in 24.2% (n=15) of the cases. EBV was identified in 58.06% (n=36) of the tumor cells nuclei, 58.06% (n=36) in the tumor cell cytoplasm, 16.12% (n=10) in tumor infiltrative lymphocytes, and 53.2% (n=33) in normal parenchymal follicle epithelial cells (NPFEC). In the T2 stage, the rate of EBV nuclear positivity in patients was significantly higher (p=0.034). The classic variant of papillary carcinoma was accompanied by a significantly higher rate of EBV-negative NPFEC (67.6%, p=0.049). In multifocal tumors, EBV positivity was found to be significantly higher in lymphocytes in the surrounding tissues (58.3%, p=0.034). CONCLUSION: A significant increase in EBV positivity in the surrounding tissue lymphocytes was observed in multifocal PTC developing on a background of HT. This suggests a possible association between HT and EBV.

Human papillomavirus infection and papillary squamous cell carcinoma in the head and neck region.

Papillary squamous cell carcinoma (PSCC) is a rare variant of SCC in the head and neck region. The role of human papillomavirus (HPV) infection in PSCC is still unclear. We retrospectively reviewed 11 PSCCs in our institute over a 21-year period and compared the HPV status of PSCCs with 26 squamous cell papillomas (SCPs). Polymerase chain reaction (PCR) amplification to detect HPV DNA and in situ hybridization (ISH) were performed to analyze the relationship between the papillary lesions and HPV infection. Immunohistochemical (IHC) staining for p16 protein expression was used to analyze the PSCC specimens. Nine of 11 (82 %), eight of 11 (73 %), and eight of 11 (73 %) PSCC samples were found to be HPV positive by PCR, ISH, and IHC staining for p16 protein expression, respectively. PSCC had a significantly higher rate of HPV infection than SCP by PCR (p = 0.002) and ISH (p = 0.001) analysis. This study presents different HPV status in two papillary neoplasms and may help to clarify the unique morphological and biological characteristics of head and neck PSCC.

A male patient with malignant lymphoma and thyroid papillary carcinoma after pediatric renal transplantation.

A 6-year-old boy received renal transplantation and was treated with methylprednisolone, cyclosporine A and mizoribine. He developed Epstein-Barr virus-associated malignant lymphoma at 10 years and thyroid papillary carcinoma at 20 years of age. Chemotherapy for the malignant lymphoma was done after withdrawal of cyclosporine A and mizoribine, and thyroidectomy was performed for thyroid carcinoma. He was well and his serum creatinine was 1.0 mg/dl at 22 years of age. To our knowledge, no pediatric renal transplant recipient who had thyroid carcinoma or two different types of tumor has been reported in Japan.

Detection of human papillomavirus in a urothelial carcinoma mimicking urethral caruncle.

Primary carcinoma of the female urethra is an uncommon diagnosis, accounting for less than 0.02% of all carcinomas in women. Urothelial carcinomas occupying the distal urethra in young females are considered to be extremely rare. Here we report what we believe to be the sixth case of primary urothelial carcinoma in the published English-language literature. The patient, a 26-year-old woman, presented with a distal urethral lesion that resembled a caruncle, but which was proved to be a urothelial carcinoma on histopathological examination of the resected specimen. Human papillomavirus type 51 DNA was detected in the tumor by polymerase chain reaction and in situ hybridization. These findings suggest that human papillomavirus might be involved in a subset of urothelial carcinomas of the urethra.

Bovine papillomavirus E5 oncoprotein upregulates parkin-dependent mitophagy in urothelial cells of cattle with spontaneous papillomavirus infection: A mechanistic study.

This study aimed to provide mechanistic insights into mitophagy pathway associated with papillomavirus infection in urothelial cells of cattle. The elimination of mitochondria via autophagy, termed mitophagy, is an evolutionarily conserved mechanism for mitochondrial quality control and homeostasis. PINK1/parkin-mediated mitophagy, a ubiquitin-dependent selective autophagy of dysfunctional mitochondria, has been described here, for the first time, in urothelial cells from 25 bladder cancers in cattle infected by bovine papillomavirus (BPV). The expression of BPV-2 and BPV-13 E5 oncoprotein was detected by RT-PCR. Abnormal mitochondria delimited by expanding phagophores, were peculiar ultrastructural features of neoplastic urothelial cells. High levels of mitochondrial phosphorylated PINK1/parkin were observed in neoplastic urothelial cells infected by BPVs. Phosphoparkin interacted with mitofusin 2 (Mfn2) and ubiquitin (Ub), which confirmed that Mfn2 is a parkin receptor at the mitochondrial level, where parkin interacted also with Ub. Furthermore, parkin established a complex that was comprised of optineurin, p62, LC3, laforin, and embryonic stem cell-expressed Ras (ERAS), that interacted with BPV E5 oncoprotein, and Bag3, which, in turn, regulated the formation of a complex composed of Hpc70/Hsp70, CHIP, an HSC70-interacting E3 ubiquitin ligase. It is conceivable that ERAS is involved in mitophagosome maturation via phosphatidylinositol 3-kinase (PI3K) pathway. Bag3, in association with Hsc70/Hsp70, may contribute to the transport and degradation of CHIP-ubiquitinated cargo as this complex recognises ubiquitinated cargos and transports them to aggresomes to be degraded. Furthermore, Bag3 may be involved in mitophagosome formation as it interacted with synaptopodin 2, which is known to play a role in mitophagosome biogenesis.

Detection of human papillomavirus type 11 in a case of Buschke-Löwenstein tumor.

We here report a 31-year-old male affected by a papillary tumor in his pubic region. At 26 years of age, he consulted a nearby clinic and was prescribed a topical cream. Although the condition was not relieved, he left the disease untreated. The gradually growing tumor adversely affected his quality of life, and he consulted another clinic, where he was referred to our hospital for surgery. The tumor had infiltrated the tissue at the base of the penis, but not the glans. After careful examination, we performed local excision of the tumor and a split-thickness skin graft. On pathological examination, elongation of the epidermis and koilocytes in the uppermost portion of the spinous layer were observed. Moreover, PCR examination confirmed the presence of human papillomavirus (HPV) type 11 in the tumor tissue. These findings supported a diagnosis of Buschke-Löwenstein tumor (BLT).

Review article: relationship of human papillomavirus with papillary squamous cell carcinoma of the upper aerodigestive tract: a review.

The purpose of this review is to evaluate case reports of papillary squamous cell carcinoma (PSCC) of the upper aerodigestive tract (UADT) to assess its relationship with human papillomavirus (HPV). The medical literature was searched for case reports of this condition. A total of 115 cases of PSCC were found that described the condition in sufficient detail. HPV detection was performed in only 22 of the 115 cases of PSCC (19%), and 11 of the 22 cases (50%) are related to this virus. The majority of cases related to HPV are produced by low-risk HPV type 6 followed by high-risk HPV type 16. Today, the association of HPV with PSCCs seems unclear because in the majority of patients tests were not performed for the detection of the HPV. This association should be clearly established to make a correct diagnosis and propose the best therapeutic strategies, such as new vaccines.

Bovine papillomavirus E5 oncoprotein binds to the activated form of the platelet-derived growth factor beta receptor in naturally occurring bovine urinary bladder tumours.

Studies regarding the functions of the bovine papillomavirus (BPV) E5 oncoprotein in vivo are lacking and no E5-mediated mechanism underlying epithelial carcinogenesis is known. We have shown that BPV-2 DNA is present in the majority of naturally occurring urinary bladder tumours of cattle and that E5 is expressed in the cancer cells. Here we show that the interaction between the platelet-derived growth factor (PDGF) beta receptor and BPV E5, described in vitro in cultured cells, takes place in vivo in bovine urinary bladder cancers. In these cancers, E5 and PDGF beta receptor colocalize, as shown by confocal microscopy, and physically interact, as shown by coimmunoprecipitation. Furthermore, the PDGF beta receptor associated with E5 is highly phosphorylated, suggesting the functional activation of the receptor upon E5 interaction. Our results demonstrate, for the first time, that E5-PDGF beta receptor interaction occurs during the natural history of bovine urinary bladder tumours, suggesting an important role for E5 in carcinogenesis. Finally, the system provides a suitable animal model of papillomavirus-associated cancer to test therapeutic vaccination against E5. Successful bladder tumour regression would provide a valuable model for therapeutic vaccination against papillomavirus-associated tumours.

Detection of human papillomavirus DNA by DNA chip in breast carcinomas of Korean women.

It remains unclear whether there is an association between human papillomavirus (HPV) infection and human breast cancer. The aim of this study was to investigate the presence of HPV DNA in breast carcinomas of Korean women and to examine the possible association between HPV and breast cancer development. For this purpose, HPV DNAs from 154 patients, including 123 patients with breast carcinoma and 31 with intraductal papilloma, and nipple tissue from 27 cancer patients were examined using the DNA chip method. HPV DNA was detected in 8 breast carcinomas (6.5%) but in no intraductal papilloma. All detected HPV genotypes were of high-risk groups. There was a slightly increased incidence in papillary carcinomas (11.5%) and invasive ductal carcinomas with adjacent intraductal papillomas (11.8%) compared to the other histological subtypes (3.2-4.3%), although the difference was not statistically significant (p = 0.126). The presence of HPV DNA was not correlated with specific prognostic predictors of disease. High-risk HPV DNA sequences were present in 6.5% of Korean patients with breast tumors. However, this study could not demonstrate whether or not such HPVs directly contribute to the development of breast cancer.

Expression of Epstein-Barr virus in renal cell carcinoma.

There have been few studies regarding the etiology of renal cell carcinoma. To examine the possible involvement of Epstein-Barr virus (EBV) in this disease, 9 renal cell carcinoma (RCC), 2 nephroblastoma (Wilms' tumor) and 2 RCC cell lines were subjected to mRNA in situ hybridization and indirect immunofluorescence staining. Messenger RNA in situ hybridization using BamHIW, EBNA LP, EBNA 2 and EBER1 probes of EBV revealed signals in all the examined samples, although some samples showed weak signals using the EBNA LP probe. Indirect immunofluorescence staining using anti-EBNA LP, anti-EBNA2, anti-LMP1 and anti-BZLF1 antibodies showed definitive fluorescence. PCR also revealed EBV DNA in all 8 RCC specimens including 7 cases other than hybridization and fluorescence. EBV infected all the RCC and nephroblastoma irrespective of the histological or clinical stage. On the other hand, EBV expression was stronger in papillary and clear cell-type RCC than chromophobe cell-type, as well as being stronger in the higher grades of RCC. These results suggest that the expression of EBV may be involved in the pathogenesis of RCC and nephroblastoma.

Evaluation of the presence of Epstein-Barr virus (EBV) in Iranian patients with thyroid papillary carcinoma.

BACKGROUND AND PURPOSE: Papillary thyroid carcinoma (PTC) is the most common thyroid cancer. EBV is one of the most important viruses related to different types of malignancies. This study investigated the relationship between EBV and papillary thyroid carcinoma. MATERIAL AND METHODS: In this study the presence of Epstein-Barr Nuclear Antigen 1 (EBNA1) gene in papillary thyroid carcinoma tissues were examined by nested-PCR method. Paraffin-embedded tissues (N=41) blocks of thyroid cancer were used. DNA was extracted from all samples and then samples were evaluated for the presence of EBV gene. RESULTS: In 41 samples, EBNA1 was detected in 65.8% of patients with papillary thyroid carcinoma which was significantly higher in younger ages. CONCLUSION: The significant presence of EBV genome in papillary thyroid carcinoma suggests that this virus may play a role in this cancer especially in younger ages. As a result, monitoring of patients with EBV latent infection for PTC can be very important.

Human Papillomavirus DNA and P16INK4A are not detected in renal tumors with immunohistochemistry and signal-amplified in situ hybridization in paraffin-embedded tissue.

The association between human papillomavirus (HPV) infection and carcinogenesis has long been established in literature, with the strongest evidence for its role in cervical carcinoma. The role of HPV in urological tumors has been investigated and sporadic reports have linked HPV infection to bladder, prostate, renal, penile, and testicular cancer. Although less rigorously studied, there are a few conflicting results about the role of HPV in the development of malignant renal tumors. Moreover, no data are available for association of HPV DNA and expression of P16 in benign renal tumors. Formalin-fixed, paraffin-embedded tissues from 62 renal tumors (40 clear cell, 9 papillary, and 3 chromophobe renal cell carcinomas, 1 collecting duct carcinoma, 2 urothelial carcinoma of renal pelvis and 7 oncocytomas) were immunostained with low-risk and high-risk HPV DNA (6, 11, 16, 18, 31, 33, 42, 51, 52, 56, 58). Tissue microarray sections of 62 tumors were stained with P16 by immunohistochemistry. Signal amplified colorimetric in situ hybridization was performed on microarray sections using biotinylated probes for HPV subtypes 6, 11, 16, 18. A nuclear dot-like signal was considered positive for low-risk and high-risk HPV by immunohistochemistry and in situ hybridization and nuclear or cytoplasmic staining is considered positive for P16. No staining for HPV DNA and P16 was found in any type of renal tumors. Our results support that HPV does not seem to play a role in the development of benign and malignant renal tumors.

Simian virus 40-like DNA sequences in human papillary thyroid carcinomas.

Sequences of the SV40 virus, a virus of Asian macaques, have been found in human tumors, such as pleural mesotheliomas, ependimomas and choroid plexus tumors. Transgenic mice carrying the SV40 large T gene under the transcriptional control of the thyroglobulin gene promoter, develop thyroid dedifferentiation and follicular thyroid cell proliferation, leading to thyroid hyperplasia and adenocarcinomas. On these bases we investigated the presence of SV40 DNA sequences in 69 samples of papillary thyroid carcinomas (PTC) and in other thyroid and non-thyroid carcinomas, as well as in benign thyroid diseases. By Southern blot and PCR amplification followed by sequence analysis, we found the presence of SV40-related sequences integrated in the tumoral DNA of three cases of PTC. At least the 203 bp fragment of the aminoterminus of large T antigen, the 294 bp fragment of the VP1 gene and the 483 bp entire regulatory region were present in the tumoral DNA of these patients. SV40 sequences were not found in tissues other than PTC. Our results demonstrate that, in addition to previous findings in mesotheliomas and brain tumors, SV40 is somehow linked to papillary thyroid carcinoma. Although our data do not demonstrate a causative role in the development of PTC, this possibility must be considered and requires further studies.

Human papillomavirus-related squamous cell carcinoma of the anal canal with papillary features.

Human papillomavirus (HPV) related squamous cell carcinoma (SCC) involving the anal canal is a well-known carcinoma associated with high-risk types of HPV. HPV-related SCC with papillary morphology (papillary SCC) has been described in the oropharynx. We describe, for the first time, a case of anal HPV-related squamous carcinoma with papillary morphology. The tumor arose from the anal mucosa. The biopsies revealed a superficially invasive SCC with prominent papillary features and associated in situ carcinoma. The tumor cells were positive for p16 and were also positive for high-risk types of HPV using chromogenic in situ hybridization. The findings are consistent with a HPV-related SCC of the anal canal with papillary features. This tumor shows histologic features similar to a papillary HPV-related SCC of the oropharynx. Additional studies are needed to characterize these lesions.

Expression of Epstein-Barr virus in thyroid carcinoma correlates with tumor progression.

There have been few studies regarding cancer progression from differentiated thyroid carcinoma to the undifferentiated one. To examine the possible involvement of Epstein-Barr virus (EBV) in this progression, 10 papillary carcinomas and 11 undifferentiated carcinomas were subjected to mRNA in situ hybridization, indirect immunofluorescence staining, polymerase chain reaction (PCR), and reverse-transcriptase PCR. mRNA in situ hybridization using a BamHIW probe revealed signals in all of the examined samples, although the signal strength was weaker in the papillary carcinomas than in the undifferentiated carcinomas. EBV nuclear antigen-2 (EBNA2) in situ hybridization produced almost the same results; however, the signals were detected less frequently in the papillary carcinomas. Indirect immunofluorescence using anti-EBNA2, anti-latent membrane protein-1 (LMP1), and anti-BZLF1 antibodies also showed positive results with high frequency and with more prominent fluorescence in undifferentiated carcinomas than in papillary carcinomas. An examination of thyroid carcinoma cell lines also confirmed these findings. EBV infected all of the thyroid carcinomas irrespective of the degree of pathological differentiation. The expression of EBV, especially of EBNA2 and LMP1 (both of which are oncogene products of EBV), was stronger in the undifferentiated carcinomas than in the papillary carcinomas. These results suggest that increased expression of EBV may be involved in the progression of thyroid papillary carcinoma to undifferentiated carcinoma.