Supaporn Suwiwat

HPV genotyping in cervical cancer in Northern Thailand: Adapting the linear array HPV assay for use on paraffin-embedded tissue

OBJECTIVES The aims of this study were to determine the prevalence of HPV infection and distribution of HPV genotypes in Northern Thai women and thereby estimate the benefit of administering the HPV vaccine in the population. METHODS Formaldehyde-fixed, paraffin-embedded samples of invasive squamous cell carcinoma from 99 patients were tested for HPV genotypes using the Linear Array HPV Genotyping Test. RESULTS HPV was detected in 96/99 (96.9%) cases. Seventy-five (78.1%) cases were single infections and 21 (21.9%) multiple. HPV16 and HPV18 were the most common subtypes, detected in 62/96 (64.4%) cases. HPV52 and HPV58 infections were found in 17/96 (17.7%) cases. Co-infection always involved HPV16. The most common co-infection was HPV16 and 52 (7 cases) but never HPV16 and 18. CONCLUSIONS Although the prevalence of HPV infection in cervical cancer of Northern Thai women is comparable to the other regions worldwide, the distribution of HPV subtypes differs with lower frequencies of HPV16 and 18, and higher frequencies of HPV52 and 58. Moreover, multiple infections are common. The vaccine against HPV16 and HPV18 can be estimated to prevent approximately two thirds of the cervical cancer cases in Northern Thailand. Although designed for use on unfixed tissue, this study shows that the Linear Array HPV Genotyping Test can be successfully used for HPV genotyping on paraffin-embedded archival tissue. This methodology also provides a means for retrospective studies on serial samples for a greater understanding of HPV genotypes, co-infections, and relationship to cervical cancer.

Prognostic significance of 14-3-3γ overexpression in advanced non-small cell lung cancer.

The 14-3-3 protein has been shown to be involved in the cancer process. However, there is no understanding of the relationship between 14-3-3γ (14-3-3 gamma) expression and prognosis in advanced non-small cell lung cancer. In this study, we therefore investigated the association between protein levels by immunohistochemistry and clinicopathological features of advanced NSCLC patients. Survival curves were estimated using the Kaplan-Meier method and tested by log-rank. Multivariate analysis was conducted with the Coxs regression model to determine independence of factors. p values less than 0.05 were considered significant. A total 153 patients were studied, with 54.3% being stage III and 45.8% stage IV. Fifty-one cases (33.3%) were squamous cell carcinomas, and 98 cases (64.1%) were adenocarcinomas. High 14-3-3γ expression was seen in 59.5% and significantly correlated with lymph node metastasis (p=0.010) and distant metastasis (p=0.017). On Kaplan-Meier analysis, high 14-3-3γ expression was associated with poorer survival with a marginal trend toward significance (p=0.055). On multivariate analysis, age, treatment, and 14-3-3γ expression proved to be independent prognostic parameters. In vitro experiments indicated that 14-3-3γ overexpression also played a potential role in cancer invasion. In conclusion, our data suggest that 14-3-3γ overexpression is associated with invasion and a poor prognosis. Therefore, 14-3-3γ may be a potential prognostic marker of advanced non-small cell lung cancer.

Prevalence of p53 mutations and protein expression in esophageal cancers in Southern Thailand

To investigate p53 alterations in esophageal squamous‐cell carcinomas of patients in the high‐risk area of southern Thailand, 72 paraffin‐embedded samples were analyzed immunohistochemically for p53 protein expression and 16 frozen samples for p53 mutational status. Forty‐two of the 72 tumors (58.3%) showed p53 protein accumulation in the nuclei of tumor cells. Expression of p53 in tumors was not significantly correlated with gender, histological grading, depth of invasion, node involvement, smoking or alcohol consumption. Analysis of the p53 gene in a sub‐set of 16 tumors showed mis‐sense mutations in 7 out of 11 p53‐positive and 1 out of 5 p53‐negative tumors. The p53 mutational spectrum was 50% transitions (3 C‐to‐T and 1 G‐to‐A, all occurring at CpG dinucleotide sites) and 50% transversions (one each, C‐to‐G, G‐to‐T, T‐to‐G, and T‐to‐A). Our findings support the hypothesis that alterations of p53 are involved in the carcinogenesis of most squamous‐cell carcinomas of the esophagus, irrespective of the population and the factors responsible for carcinogenesis. The mutation profile of the p53 gene might indicate etiologic contributions of different mutagen exposures in patients from high‐risk areas of southern Thailand. Int. J. Cancer 72:23–26, 1997.

Prognostic Value of HPV18 DNA Viral Load in Patients with Early-Stage Neuroendocrine Carcinoma of the Uterine Cervix

OBJECTIVES To evaluate the clinicopathologic correlation and prognostic value of HPV18 DNA viral load in patients with early-stage cervical neuroendocrine carcinoma (NECA). METHODS Formalin-fixed, paraffin- embedded tissue of cervical NECA patients with known HPV18 infection and clinicopathologic data including follow-up results were collected. The HPV18 DNA load was assessed with quantitative PCR targeting the HPV18 E6E7 region. RESULTS Twenty-one patients with early-stage (IB-IIA) cervical NECA were identified. HPV18 DNA viral load ranged from 0.83 to 55,174 copies/cell (median 5.90). Disease progression, observed in 10 cases (48%), was not significantly associated with any clinicopathologic variables. However, the group of patients with progressive disease tended to have a higher rate of pelvic lymph node metastasis (50% versus 9%, p=0.063) and a lower median value of HPV18 DNA viral load (4.37 versus 8.17 copies/cell, p=0.198) compared to the non-recurrent group. When stratified by a cut-off viral load value of 5.00 copies/cell, the group of patients with viral load ≤5.00 copies/cell had a significantly shorter disease-free survival than the group with viral load >5.00 copies/cell (p=0.028). The group with a lower viral load also tended to have a higher rate of disease progression (75% versus 31%, p=0.080). No significant difference in the other clinicopathologic variables between the lower and higher viral load groups was identified. CONCLUSION THPV18 DNA viral load may have a prognostic value in patients with early-stage NECA of the cervix. A low viral load may be predictive of shortened disease-free survival in these patients.

Hepatic cytotoxic T-cell infiltrates in patients with peripheral T-cell proliferative diseases/lymphomas: Clinicopathological and molecular analysis

Seventy patients with various types of peripheral T‐cell proliferative disease/lymphoma who manifested with prolonged fever, weight loss, anemia, lymphadenopathy, hepatosplenomegaly and elevated serum levels of alkaline phosphatase and/or lactate dehydrogenase were evaluated. Histopathological examination of the livers revealed T‐cell infiltration into the hepatic sinusoids and portal tracts. The morphology of the infiltrated T cells varied from mature small lymphocytes to malignant lymphoid cells. The liver pathology was classified into four groups on the basis of cellular atypia. Group A and group B showed mature lymphoid cell infiltration; however, only group B had multiple large areas of hepatocellular necrosis. Group C showed atypical lymphoid cell infiltration and in group D malignant lymphoid cell infiltrates were demonstrated. The majority of the antigenic phenotypes of these T‐cell infiltrates were CD3+, CD4–, CD8+, CD20–, CD45RO+, CD56–, CD57–, TIA‐1+ and βF1–. Epstein–Barr virus RNA in the nuclei of the infiltrated T cells was recorded in 38.6% of the patients and was more common in groups C and D. Patients in groups B, C and D had a very poor prognosis, median survival was only 1 month, whereas median survival in group A patients was 36 months. Chemotherapy was not effective in improving survival.  Monoclonal  band/s  of  T‐cell  receptors  (TCR) β and/or γ gene rearrangements were detected in 88.6% of patients, and DNA‐sequence analysis showed high identity to the human TCR germline gene.

Methylation of 14‑3‑3σ gene and prognostic significance of 14‑3‑3σ expression in non‑small cell lung cancer

Loss of 14-3-3σ expression through DNA methylation has been associated with carcinogenesis and the prognosis for various cancer types. Detection of methylation of the gene in serum may be useful for diagnostic utility. The present study aimed to investigate the correlation between 14-3-3σ methylation level in 36 paired tumor tissues of non-small cell lung cancer (NSCLC) and matched serum using methylation-specific polymerase chain reaction. The prognostic significance of 14-3-3σ expression in 167 NSCLC was also evaluated using immunohistochemistry. Methylation of the 14-3-3σ gene was identified in all samples. The methylation level in the serum (mean 87.7%, range 64.6–100%) was higher compared with tumor (mean 46.7%, range 25.3–56.3%). However, no significant correlation between methylation levels in tissues and serums was observed (Spearmans correlation, −0.036; P=0.837). In the 167 tumor tissues, the majority of the cases (83.8%) exhibited negative expression. Adenocarcinoma is more likely to exhibit negative expression (91.4%) compared with squamous cell carcinoma (70.2%). No significant difference was identified in the overall survival according to 14-3-3σ expression status and 14-3-3σ expression did not demonstrated independent prognostic significance. In conclusion, NSCLC harbors certain levels of 14-3-3σ methylation in the tumor and the sera of patients. The clinical value of serum 14-3-3σ methylation should be further elucidated. Immunohistochemical expression 14-3-3σ protein has limited value on prognostic significance.

Epstein-Barr virus-associated smooth muscle tumor of the tonsil.

Smooth muscle tumors of the tonsil are rare. Recently, the occurrence of Epstein-Barr virus-associated smooth muscle tumor (EBV-SMT) has been increasingly recognized in immunocompromised patients, mainly post-transplantation and AIDS patients. The clinicopathologic features of EBV-SMT are different from conventional smooth muscle tumors. To the best of our knowledge, EBV-SMT involving the tonsil in an AIDS patient has not been reported. A 27-year-old man presented with a 2.2cm right tonsillar mass six months after AIDS diagnosis. The tumor was composed of a cellular proliferation of oval to spindle-shaped cells with mitotic count up to 10 in 10 high-power fields. The diagnosis of EBV-SMT was confirmed by in situ hybridization for EBV-encoded RNA (EBER) transcripts. Synchronous lesions were also detected in the liver and peritoneum by an abdominal computed tomographic scan. EBV-SMT should be included in the differential diagnoses of a mesenchymal tumor in immunocompromised patients, and in the differential diagnoses of a smooth muscle tumor occurring in uncommon sites including the tonsil.

Erythema multiforme-like cutaneous lesions in monomorphic epitheliotropic intestinal T-cell lymphoma: a rare case report

Monomorphic epitheliotropic intestinal T‐cell lymphoma (MEITL), also known as Type II enteropathy‐associated T‐cell lymphoma (EATL), is an aggressive peripheral T‐cell lymphoma. EATL generally presents in adults with gastrointestinal symptoms. Skin involvement is very rare, found only in approximately five percent of patients. The authors report a 67‐year‐old Asian male who presented with chronic diarrhea and developed erythema multiforme‐like cutaneous lesions. A skin biopsy revealed extensive pagetoid spread of atypical lymphocytes in the epidermis. The results of an immunohistochemistry test led to a diagnosis of MEITL. This report points to the need for dermatologists and dermatopathologists to consider a possible diagnosis of MEITL when encountering similar cases.