Cervical Intraepithelial Neoplasia Specimens Research Articles

FHIT and C-MYC expression in cervical histology and cytology as biomarkers for detecting high-grade intraepithelial neoplasia in human papillomavirus-positive women.

BACKGROUND: The current cervical cancer screening strategies based on Papanicolaou (Pap) and Human papillomavirus (HPV) tests receive great achievement but still exhibit many limitations in clinical practice. Exploring new biomarkers as stratified management method in HPV primary screening is becoming the tendency of current research.METHODS: Immunocytochemistry (ICC) of FHIT and C-MYC were performed on exfoliated cervical cells from 197 eligible high-risk HPV positive women. Mann-Whitney U test, Pearson Chi-Square test, logistic regression analysis and receiver operating characteristic (ROC) curves were used to assess the diagnostic efficiency.RESULTS: ICC staining intensity of FHIT and C-MYC in high-grade cervical intraepithelial neoplasia (CIN) specimens was significantly different from low-grade CIN and normal specimens. Compared with Pap test, ROC analysis of ICC in detecting high-grade CIN resulted in a larger area under the curve (AUC) (0.805 and 0.814 vs 0.723, 0.001). FHIT achieved higher sensitivity than Pap test (79.41% vs 66.67%, 0.04). Logistic regression analysis of the combination of two biomarkers led to higher AUC value, specificity and PPV than any single biomarker.CONCLUSIONS: The utility of FHIT and C-MYC ICC analysis in cervical exfoliated cells of HPV-positive women displayed superior diagnostic potential and may improve clinical performance of cervical cancer screening.

Correlation between ER, PR, P53, Ki67 Expression and High-Risk HPV Infection in Patients with Different Levels of Cervical Intraepithelial Neoplasia

To investigate the correlation between high-risk human papillomavirus infection and the expression of ER, PR, P53 and Ki67 in patients with different grades of cervical intraepithelial neoplasia. A retrospective study was conducted in which from June 2016 to June 2018, 140 specimens of cervical intraepithelial neoplasia were collected from the pathology department of the Beijing Tongren Hospital that included 40 specimens of cervical intraepithelial neoplasia1, 50 specimens of cervical intraepithelial neoplasia2 and 50 specimens of cervical intraepithelial neoplasia1. The expression of ER, PR, p53 and Ki67 were determined by immunohistochemistry. High-risk human papillomavirus infections were detected by polymerase chain reaction fluorescence quantification and were given the correlation analysis. Results of the 140 specimens indicated the rates of human papillomavirus16 and human papillomavirus18 in cervical intraepithelial neoplasia1 specimens were 27.5 and 25.0 % respectively, in cervical intraepithelial neoplasia2 specimens were 64.0 and 60.0 %, respectively and in cervical intraepithelial neoplasia3 specimens were 90.0 and 92.0 % respectively, the difference were statistically significant (p<0.05). There was no significant correlation between human papillomavirus 16 and human papillomavirus 18 positive rate and patient age, tissue differentiation, and tumour size. With the increasing cervical intraepithelial neoplasia grade, the rate of ER, PR, p53 and Ki67 expression in specimens was also increased significantly (p<0.05). Pearson correlation analysis showed there were positive correlation between the rates of human papillomavirus 16 and human papillomavirus 18 and the rates of ER, PR, p53 and Ki67 expression (p<0.05). In conclusion with increasing cervical intraepithelial neoplasia level, rate of high-risk human papillomavirus infection increased along with increased rate of expression of ER, PR, p53 and Ki67.

RETRACTED: Correlation between ER, PR, P53, Ki67 expression, and high-risk HPV infection in patients with different levels of cervical intraepithelial neoplasia

This study was designed to investigate the correlation between high-risk human papillomavirus (HPV) infection and the expression of IHC markers (ER, PR, p53, Ki67) in patients with different grades of cervical intraepithelial neoplasia (CIN). It was a retrospective study, which was conducted from June 2016 to June 2018. 140 specimens of CIN were collected from the pathology department of a certain hospital that included 40 specimens of CIN1, 50 specimens of CIN2 and 50 specimens of CIN3. The expression of ER, PR, P53 and Ki67 were determined by immunohistochemistry. The high-risk HPV infections were detected by PCR fluorescence quantification and were given the correlation analysis. In the 140 specimens, the positive rates of HPV16 and HPV18 in CIN1 specimens were 27.5% and 25.0% respectively, and in CIN2 specimens were 64.0% and 60.0% respectively, and in CIN3 specimens were 90.0% and 92.0% respectively, the difference were statistically significant (p&lt;0.05). There were no significant correlation (p&lt;0.05) between HPV16 and HPV18 positive rate and patient age, tissue differentiation, and tumor size. With the increased of CIN grade, the positive rate of ER, PR, P53 and Ki67 expression in specimen were also increased significantly, and the difference were statistically significant (p&lt;0.05). Pearson correlation analysis showed there were positive correlation (p&lt;0.05) between the positive rates of HPV16 and HPV18 and the positive rates of ER, PR, P53 and Ki67. With the increase of CIN level, the positive rates of high-risk HPV infection as well as ER, PR, P53 and Ki67 are increased, and they have positive correlation.

Comprehensive genomic variation profiling of cervical intraepithelial neoplasia and cervical cancer identifies potential targets for cervical cancer early warning

BackgroundTo better understand the pathogenesis of cervical cancer (CC), we systematically analysed the genomic variation and human papillomavirus (HPV) integration profiles of cervical intraepithelial neoplasia (CIN) and CC.MethodsWe performed whole-genome...

Comprehensive Genomic Variation Profiling of Cervical Intraepithelial Neoplasia and Cervical Cancer Identifies Potential Targets for Cervical Cancer Early Warning

To better understand the pathogenesis of cervical cancer (CC), we systematically analyzed the genomic variation and human papillomavirus (HPV) integration profiles of cervical intraepithelial neoplasia (CIN) and CC. We performed whole-genome sequencing (WGS) or whole-exome sequencing (WES) of 102 tumor-normal pairs and human papillomavirus (HPV) probe capture sequencing (HPCS) of 45 CCs, 44 cervical intraepithelial neoplasia (CIN) samples, and 25 normal cervical samples, and constructed strict integrated workflow of genomic analysis. Mutational analysis identified eight significantly mutated genes in CC including four genes (FAT1, MLL3, MLL2, and FADD), which have not previously been reported in CC. Targetable alterations were identified in 55.9% of patients. In addition, HPV integration breakpoints occurred in 97.8% of the CC samples, 70.5% of the CIN samples and 42.8% of the normal cervical samples with HPV infection. Integrations of high-risk HPV strains in CCs, including HPV16, 18, 33 and 58, also occurred in the CIN samples. Moreover, gene mutations were detected in 52% of the CIN specimens, and 54.8% of these mutations occurred in genes that also mutated in CCs. Our results lay the foundation for a deep understanding of the molecular mechanisms and finding new diagnostic and therapeutic targets of CC. Funding Statement: This work was supported by grants from the National High Technology Research and Development Program of China (2012AA02A205), the Chinese National Key Program on Basic Research (2014CB965002), the National Natural Science Foundation of China (81272306 and 81472639), and the Shanghai Commission for Science and Technology (15431902900 and 15JC1403000). Declaration of Interests: The authors declare that they have no competing interests. Ethics Approval Statement: Chinese CC patients were recruited under the approval of the Research Ethics Committee of Southwestern Hospital (No. 2014-016) and this project and protocols were conducted according to the Declaration of Helsinki. Informed consents were provided.

Immunohistochemical Expression of Bronchial Dysplasia Associate Proteins in Uterine Cervical Lesions [6P

INTRODUCTION: Bronchial dysplasia associate proteins are involved in the proliferation, cell differentiation and adhesion, tumorigenesis, invasion and metastasis of cancer. We examined the immunohistochemical expression of bronchial dysplasia associate proteins in uterine cervical lesions. METHODS: Specimens from 32 patients with uterine cervical intraepithelial neoplasia (CIN) or uterine cervical invasive carcinoma were immunohistochemically evaluated for the expression of the 6 proteins associated with bronchial dysplasia: serpin peptidase inhibitor B2 (SERPINB2), carboxypeptidase A4 (CPA4), keratin 6A and 13, interleukin 1 receptor antagonist (IL-1Ra), small proline-rich protein 3 (SPRR3) and desmoglein 3 (Dsg3). Immunohistochemistry staining intensity was quantitatively scored from 0 to 2+. This study was approved by the Institutional Review Board. RESULTS: The positive expression rates of SERPINB2, CPA4, Dsg3 and SPRR3 were 84%, 62.5%, 27%, and 18.9% in the uterine cervical cells, 87.5%, 75%, 70.3%, and 0% in the biopsy tissue specimens, respectively. The expression of keratin6A/13 was not specific, and the expression of IL-1Ra only in the cervical dysplastic cells revealed positive, but negative in normal cells and cervical invasive carcinoma cells, and in all biopsy tissues. The expression of SERPINB2 seemed to have positive correlation with the progression of lesions, but the expression of CPA4 in only the dysplastic cells revealed positive. CONCLUSION: SERPINB2 might be useful as a marker for uterine cervical lesions because it was expressed according to the grade of the CIN and cervical invasive carcinoma specimens. CPA4, Dsg3, and IL-1Ra could be potential immunohistochemical markers for uterine cervical lesions because of positive expression in the dysplastic cells.

HMGB1 overexpression correlates with poor prognosis in early-stage squamous cervical cancer.

High mobility group box 1 (HMGB1) is associated with tumor progression and a poor prognosis; microtubule-associated protein 1 light chain 3 (LC3) plays a critical role in autophagy. However, the roles of HMGB1 and LC3 in squamous cervical cancer (SCC) remain unclear. An array of 166 early-stage SCC, 62 cervical intraepithelial neoplasia (CIN), and 50 normal cervical tissue samples was assessed. HMGB1 and LC3 protein levels were examined by immunohistochemistry, and the associations of HMGB1 and LC3 levels with clinicopathological characteristics evaluated, to assess their prognosis significance. High nuclear HMGB1 levels were detected in 72.9% SCC cases; 16% cases showed cytoplasmic expression of HMGB1 in cancer cells with low nuclear expression. Interestingly, HMGB1 levels in SCC samples were significantly higher than CIN and control specimens, while lower LC3 expression was found in SCC samples (P < 0.001). Nuclear HMGB1 expression was weakly negatively correlated to LC3 amounts (r = -0.254, P = 0.001). High nuclear HMGB1 levels were associated with vascular metastasis (P < 0.05). In addition, cytoplasmic HMGB1 expression was associated with lymph node metastasis (P < 0.05). Furthermore, high nuclear HMGB1 levels and cytoplasmic HMGB1 expression predicted poor overall survival (OS) and disease-free survival (DFS). Meanwhile, high LC3 expression was associated with favorable prognosis. Multivariate analysis showed that both nuclear and cytoplasmic HMGB1 expressions were independent prognostic factors for overall- and disease-free survival, along with nodule metastasis. HMGB1 overexpression plays a significant role in SCC progression. Both nuclear and cytoplasmic HMGB1 are independent factors for poor prognosis in early-stage SCC.

Immunohistochemical Expression of Serpin Peptidase Inhibitor B2 and Carboxypeptidase A4 in Uterine Cervical Lesions [200

OBJECTIVE: Serpin peptidase inhibitor B2 is involved in the invasion and metastasis of cancer. Carboxypeptidase A4 may have a correlation with the grade of prostate cancer. We examined the immunohistochemical expression of serpin peptidase inhibitor B2 and carboxypeptidase A4 in uterine cervical lesions. MATERIALS AND METHODS: Specimens from 25 patients with uterine cervical intraepithelial neoplasia (CIN) or uterine cervical invasive carcinoma were immunohistochemically evaluated for the expression of the two markers, serpin peptidase inhibitor B2 and carboxypeptidase A4. Of the patients from whom uterine cervical cells and biopsy tissue specimens were obtained, eight had normal tissues (32%), two had CIN1 lesions (40%), eight had CIN2 lesions (32%), five had CIN3 lesions (20%), and two had cervical invasive carcinoma (8%). Immunohistochemistry staining intensity was quantitatively scored from 0 to 2+. This study was approved by the institutional review board. RESULT: The positive expression rates of serpin peptidase inhibitor B2 and carboxypeptidase A4 were 75% and 20% in the uterine cervical cells and 63% and 0% in the biopsy tissue specimens, respectively. The expression of serpin peptidase inhibitor B2 seemed to have a positive correlation with the progression of lesions, but the expression of carboxypeptidase A4 in the only dysplastic cells revealed positive. The expression of serpin peptidase inhibitor B2 was positive, but that of carboxypeptidase A4 was negative in cases of cervical invasive carcinoma. CONCLUSION: Serpin peptidase inhibitor B2 might be useful as a marker for uterine cervical lesions because it was expressed according to the grade of the CIN and cervical invasive carcinoma specimens. Carboxypeptidase A4 could be potential immunohistochemical markers for uterine cervical lesions because of positive expression in the dysplastic cells.

Molecular cytogenetic analyses of hTERC (3q26) and MYC (8q24) genes amplifications in correlation with oncogenic human papillomavirus infection in Czech patients with cervical intraepithelial neoplasia and cervical carcinomas.

It is known that cervical cancer develop from precancerous intraepithelial neoplasia (CIN) which is characterized by series of genetic abnormalities. The progression of CIN to cervical carcinoma has been associated especially with the genomic integration of oncogenic human papilloma virus (HPV) and gain of the human telomerase RNA gene hTERC (3q26) and MYC (8q24). In this study, cytology specimens of cervical intraepithelial neoplasia and cervical carcinoma from 74 Czech women were analyzed using the triple-color Cervical FISH Probe Kit designed for identification of HPV infected cells and copy number aberration of the hTERC and MYC genes. HPV-positivity exhibited 70% of patients with premalignant lesions (CIN I- CIN III, carcinoma in situ), chromosomal changes were found in 53.3% of cases - MYC amplification had 33.3% of women with CINI- CIN III and 50% with carcinoma in situ. Amplification of hTERC was detected in 16.7% of patient with CIN I, in 50% with CIN II, in 58.3% with CIN III and in 66.7% with carcinoma in situ. Based on HPV-positivity and the occurrence of chromosomal aberrations, patients were divided into high-, intermediate- and low-risk group. Among women with cervical carcinomas, HPV infection was detected in 90.1% of specimens and chromosomal aberrations were found in 87.5% of samples. Amplification of MYC gene was detected in 25% and hTERC gene in 62.5% of patients. According to the histopathological grade of tumors, MYC gene amplification occurred more frequently in specimens of spinocellular carcinoma than adenocarcinoma (p=0.029). We found no association between the frequency of cytogenetic lesions and the incidence of lymphangiogenesis or lymph node metastases in cervical carcinoma patients. Simultaneous hTERC and MYC genes amplification was significantly more frequent in samples of cervical carcinomas than in premalignant lesions (p=0.008).In acohort of 26 patients with cervical carcinoma we used oligo-based GGH+SNP microarray technique for the high resolution mapping of copy number changes of hTERC and MYC genes. We found that recurrent gain of genetic material in chromosome 3q26 area carrying hTERC gene of size 43.6 Mb between 3q25.1-3qter and duplication of 3q were the most common genomic identifications of amplified gene. In MYC locus array-CGH profiling identified duplication of 8q and trisomy 8 as frequent genomic changes.Our work confirmed that in cervical carcinoma gains of hTERC and MYC genes are specific genomic changes associated with developing of malignant phenotype. We also showed that in premalignant stages HPV-FISH assay can be used as an effective diagnostic procedure to identify patients carrying highly risking HPV infection and chromosomal aberrations associated with this malignancy. cervical cancer, cervical dysplasia, HPV infection, hTERC amplification, MYC amplification, FISH, array-CGH.

Establishment of keratinocyte colonies from small‐sized cervical intraepithelial neoplasia specimens

The size of human cervical intraepithelial neoplasia (CIN) biopsies is usually very small and standard methods do not allow an adequate number of keratinocytes to be isolated for culturing purposes. In this study, a new approach to establish keratinocyte cultures from small CIN a tissue fragments was developed. Neoplastic specimens and corresponding normal tissues, which were used as controls, were digested with collagenase. Tissue-derived fibroblasts and keratinocytes were co-cultured in calcium and serum medium. Single keratinocyte colonies from primary cultures were expanded using a culture medium optimized in our laboratory. Primary keratinocyte colonies, as well as expanded colonies, were tested for epithelial and cervical markers such as 5, 14, 17, and 19 keratins, and p63 by immunofluorescence. Our results indicate that a variable number of primary keratinocyte colonies could be detected in neoplastic cultures, depending on the grade of cervical lesions from which the colonies originated. Single colonies, when cultured with our new medium, grew at a high rate with uniform size and morphology for some passages. Epithelial and p63 markers were expressed in keratinocyte colonies, as well as in expanded colonies. In conclusion, our study reports a rapid and easy culturing system which enables keratinocyte colonies from minute cervical tumor tissues to be obtained. Moreover, using the new culture medium, keratinocyte colonies can be expanded at a high proliferative rate.

An evaluation of p16INK4a expression in cervical intraepithelial neoplasia specimens, including women with HIV-1

Although several studies have evaluated the role of p16(INK4a) as a diagnostic marker of cervical intraepithelial neoplasia (CIN) and its association with disease progression, studies regarding the role of p16(INK4a) in human immunodeficiency virus (HIV)-infected patients remain scarce. The present study was designed to determine the potential utility of p16(INK4a) as a diagnostic marker for CIN and invasive cervical cancer in HIV-positive and negative cervical specimens. An immunohistochemical analysis of p16(INK4a) was performed in 326 cervical tissue microarray specimens. Performance indicators were calculated and compared using receiving operating characteristics curve (ROC)/area under the curve. In HIV-1-negative women, the percentage of cells that was positive for p16(INK4a) expression was significantly correlated with the severity of CIN (p < 0.0001). A ROC curve with a cut-off value of 55.28% resulted in a sensitivity of 89%, a specificity of 81%, a positive predictive value of 91% and a negative predictive value of 78%. HIV-seropositive women exhibited decreased expression of p16(INK4a) in CIN2-3 specimens compared with HIV-negative specimens (p = 0.031). The ROC data underscore the potential utility of p16(INK4a) under defined conditions as a diagnostic marker for CIN 2-3 staging and invasive cervical cancer. HIV-1 infection, however, is associated with relatively reduced p16(INK4a) expression in CIN 2-3.

Dysregulated microRNAs in the pathogenesis and progression of cervical neoplasm

MicroRNAs (miRNAs) play an important role in a variety of physiological as well as pathophysiological processes, including carcinogenesis. The aim of this study is to identify a distinct miRNA expression signature for cervical intraepithelial neoplasia (CIN) and to unveil individual miRNAs that may be involved in the development of cervical carcinoma. Expression profiling using quantitative real-time RT-PCR of 202 miRNAs was performed on micro-dissected high-grade CIN (CIN 2/3) tissues and compared to normal cervical epithelium. Unsupervised hierarchical clustering of the miRNA expression pattern displayed a distinct separation between the CIN and normal cervical epithelium samples. Supervised analysis identified 12 highly differentially regulated miRNAs, including miR-518a, miR-34b, miR-34c, miR-20b, miR-338, miR-9, miR-512-5p, miR-424, miR-345, miR-10a, miR-193b and miR-203, which distinguished the high-grade CIN specimens from normal cervical epithelium. This miRNA signature was further validated by an independent set of high-grade CIN cases. The same characteristic signature can also be used to distinguish cervical squamous cell carcinoma from normal controls. Target prediction analysis revealed that these dysregulated miRNAs mainly control apoptosis signaling pathways and cell cycle regulation. These findings contribute to understanding the role of microRNAs in the pathogenesis and progression of cervical neoplasm at the molecular level.

Comparison of Clinical Performance of Abbott RealTi m e High Risk HPV Test with That of Hybrid Capture 2 Assay in a Screening Setting

Randomized trials have produced sound evidence about the efficacy of screening with human papillomavirus (HPV) DNA tests in reducing cervical cancer incidence and mortality. We evaluated the clinical performance and reproducibility of the Abbott RealTime High Risk (HR) HPV test compared with that of the HR hybrid capture 2 (HC2) assay as assessed by a noninferiority score test. A random sample of 998 cervical specimens (914 specimens of cervical intraepithelial neoplasia less severe than grade 2 [<CIN2] and 84 specimens of cervical intraepithelial neoplasia grade 2 or more severe lesions [≥CIN2]), collected in the Florence and Catania cervical Cancer Screening Programs from women aged 25 to 64 and already tested by HR HC2, were retested with the Abbott RealTime HR HPV test. Absolute specificity was 92.3% (95% confidence interval [CI], 90.4 to 94.0) and 92.6% (95% CI, 90.7 to 94.2) for the Abbott RealTime HR HPV test and the HR HC2, respectively. Absolute sensitivity was 96.4% (95% CI, 89.9 to 99.3) and 97.6% (95% CI, 91.7 to 99.7) for the Abbott RealTime HR HPV test and the HR HC2, respectively. The noninferiority score test revealed that the clinical sensitivity and specificity of the Abbott RealTime HR HPV test were not inferior (P = 0.004 and 0.009, respectively) to those of HR HC2. Overall agreement between the two assays was 96.5%, with a k value of 0.86 (CI 95%, 0.82 to 0.91). We evaluated the intralaboratory reproducibility by retesting 521 samples at least 4 weeks after the first test; the crude agreement between the first and second test was 98.5%, with an overall k value of 0.97 (CI 95%, 0.95 to 0.99). This test fully satisfies the requirements of a primary cervical cancer screening test. This assay differentiates between HPV16, HPV18, and non-HPV16/18 types in every specimen, but how to use this information in a screening setting still is unclear.

Immunolabeling of the Inhibin-βA and -βB Subunit in Normal and Malignant Human Cervical Tissue and Cervical Cancer Cell Lines

Inhibins, dimeric peptide hormones composed of an α-subunit and 1 of 2 possible β subunits (βA or βB), exhibit substantial roles in human reproduction and in endocrine-responsive tumors. However, it is still unclear whether normal and cancerous cervical tissues as well as cervical cancer cell lines express the inhibin-βA and -βB subunits. Normal human uterine cervical tissue was obtained from 4 premenopausal nonpregnant patients. In addition, a total of 32 specimens of cervical intraepithelial neoplasia (CIN) of different stages were obtained (CIN 1 = 10, CIN 2 = 9, and CIN 3 = 13). Moreover, 30 squamous cervical cancer samples of well-differentiated (grade 1; n = 10), moderate differentiated (grade 2; n = 10), and poorly differentiated (grade 3; n = 10) grading were analyzed. An immunohistochemical staining reaction for inhibin-βA and -βB subunits could be observed in normal and malignant cervical tissue as well as in cervical cancer cell lines. Regarding inhibin-βA significant differences were observed between normal tissue and CIN 1 and CIN 3. Moreover, the immunohistochemical staining reaction for inhibin-βA was significantly higher in CIN 3 compared with that in cervical carcinoma grades 1 and 2. The inhibin-βB expression was higher in CIN and cervical cancer compared with that in normal cervical tissue. Inhibin-βB was significantly higher in CIN 2 and CIN 3 compared with cancer tissues of histological grade 1. In addition, a significant increase of the staining intensity was observed between cervical cancer grades 1 and 2 as well as grade 3. Both inhibin-β subunits demonstrated a differential expression in CIN and squamous cancer, suggesting important roles in cervical carcinogenesis. Inhibin-βA might be important during progression of CIN, whereas the inhibin-βB subunit could exert a substantial function during differentiation of cervical carcinomas. Moreover, the synthesis of this subunit in cervical carcinoma cell lines also allows the use of this cell line to elucidates their functions in cervical cancer pathogenesis.

HER2/neu (c‐erbB‐2) gene amplification and protein expression are rare in uterine cervical neoplasia: a tissue microarray study of 814 archival specimens

Published studies have reported widely variable incidence of HER2/neu (c-erbB-2) protein expression and HER2/neu (c-erbB-2) gene amplification in cervical carcinoma. We examined tissue microarrays (TMAs) constructed from 814 formaldehyde-fixed paraffin-embedded archival specimens of cervical intraepithelial neoplasia (CIN)1 (n = 262), CIN2 (n = 230), CIN3 (n = 186) and invasive carcinoma (n = 136), for HER2/neu protein expression by immunohistochemistry (IHC) and for HER2/neu gene amplification by chromogenic in situ hybridization (CISH). We found moderate or strong immunohistochemical positivity for HER2/neu in 64 of 814 specimens (7.9%). Using CISH, polysomy of the HER2/neu gene was detected in 87 cases (10.7%), low/borderline amplification in five cases (0.6%) and true amplification in four cases (0.5%). The correlation between IHC and CISH was statistically significant in CIN2, CIN3 and invasive cervical carcinoma specimens. When present, Her-2/neu positivity is more commonly seen in higher grades of cervical dysplasia and in carcinoma. However, this large TMA study shows that HER2/neu oncoprotein expression and HER2/neu gene amplification overall are uncommon events in cervical neoplasia. This provides compelling evidence that HER2/neu plays no major role in the development and progression of cervical neoplasia.

Immunohistochemical evaluation of insulin-like growth factor I receptor status in cervical cancer specimens.

The insulin-like growth factor I receptor (IGF-IR) is exceptionally overexpressed in many cervical-cancer-derived cell lines. It is postulated that a decrease of p53 protein levels due to human papillomavirus (HPV) infection may contribute to the up-regulation of IGF-IR expression in cervical cancer cells because transcription of IGF-IR is strictly down-regulated by p53. To evaluate this fact in clinical cervical cancer specimens, we checked the expression levels and activated status of IGF-IR by immunohistochemistry. Formalin-fixed and paraffin-embedded specimens obtained by conization or hysterectomy were stained with anti-IGF-IR and with an antibody recognizing phosphorylated tyrosine at its c-terminus. The expression levels of IGF-IR were significantly high in cervical intraepithelial neoplasia (CIN) III and invasive cancer specimens. Phosphorylation of IGF-IR was promoted in all CIN and invasive cancer specimens, and its intensity was related to the promotion of lesions. Interestingly, IGF-IR overexpression was missing in the basal layer of CIN I and II lesions, whereas it was evenly distributed in CIN III and invasive cancer lesions. This IGF-IR overexpression pattern may be utilized in the diagnosis of HPV infection status in CIN lesions.

Expression of the p16INK4a and Ki-67 in relation to the grade of cervical intraepithelial neoplasia and high-risk human papillomavirus infection

The purposes of this study were to evaluate the expression of p16(INK4a) (referred as to p16) and Ki-67 in cervical intraepithelial neoplasia (CIN), and the correlation between high-risk human papillomavirus (HPV) infection and the above biomarkers. We analyzed 31 patients who were diagnosed with CIN at Kwandong University Myongji Hospital from October 2006 to September 2007. CIN specimens (CIN1, 12; CIN2, 6; CIN3, 13) were obtained by colposcopy-directed biopsy (CDB) or loop electrical excision procedure (LEEP). The expressions of p16 and Ki-67 were evaluated by immunohistochemical methods with antibodies to p16 and Ki67. The immunohistochemical staining results were classified into four grades: 0, 1, 2 and 3. HPV genotyping or Hybrid Capture-II test was used to detect high-risk HPV. The expression of p16 (p<0.001) and Ki-67 (p=0.003) were positively associated with CIN grade. p16 expressions increased significantly with high-risk HPV infection (p=0.014), especially HPV type 16 and 58. Ki-67 expression was not related with high-risk HPV. There was positive correlation between the expression of the p16 and Ki-67 (p=0.007). CIN grade were positively related to the expression of p16 and Ki-67. p16 expressions of high-risk HPV specimens significantly increased more than Ki-67. Therefore, in the diagnosis of CIN and high-risk HPV infection, p16 can be a useful biomarker.

Mandatory fortification with folic acid in the United States is associated with increased expression of DNA methyltransferase-1 in the cervix

Objective The objective of this study was to evaluate whether mandatory fortification of grain products with folic acid in the United States is associated with changes in DNA methyltransferase-1 (Dnmt-1) expression in cells involved in cervical carcinogenesis. Methods Archived specimens of cervical intraepithelial neoplasia (CIN) diagnosed before (1990–1992) and after (2000–2002) mandatory folic acid fortification were used to examine the expression of Dnmt-1 in specific lesions involved in cervical carcinogenesis by immunohistochemistry. The total number of lesions examined was 101 in the prefortification period and 96 in the postfortification period. Immunohistochemical staining for Dnmt-1, its assessment, and data entry were blinded with regard to the fortification status. Results Age- and race-adjusted mean percentage of cells positive for Dnmt-1 or the Dnmt-1 score was significantly higher in all lesion types (i.e., normal cervical epithelium, reactive cervical epithelium, metaplastic cervical epithelium, CIN, or carcinoma in situ) detected in the postfortification period compared with the prefortification period ( P < 0.05, all comparisons). The degree of Dnmt-1 was significantly higher ( P < 0.0001) in CIN ≥2 lesions compared with CIN ≤1 lesions, regardless of the fortification group. Conclusion These results suggest that mandatory fortification with folic acid in the United States seems to have resulted in a change in the degree of expression of Dnmt-1 in cells involved in cervical carcinogenesis. Because the approach we have taken to demonstrate these differences have limitations inherent to a study of this nature and this is the first study to report a folate fortification associated change in Dnmt-1, validating these results in other study populations and/or with other techniques of assessing Dnmt-1 will increase the scientific credibility of these findings.

The Mandatory Fortification of Foods with Folic Acid in the United States is not associated with an Improvement in the Degree or Pattern of Global DNA Methylation in the Cervical Epithelium

The purpose of this study was to evaluate the effects of folic acid fortification of foods on global DNA methylation (GDM) by comparing the methylation status in specific cells identified in archival specimens of cervical intraepithelial neoplasia (CIN) diagnosed prior to fortification (1990–92) and after fortification (2000–02). The degree of GDM was assessed by an immunohistochemical technique that used monoclonal antibodies specific for 5-methyl cytosine (5-mc) in 121 lesions identified in specimens of 39 women in the pre-fortification period and in 170 lesions identified in specimens of 49 women in the post-fortification period. The mean percentage of cells positive for 5-mc (age and race adjusted) was not significantly different between pre and post fortification periods in any of the lesion types (normal cervical epithelium, reactive cervical epithelium, metaplastic cervical epithelium, CIN 1, CIN 2, CIN 3 or carcinoma in situ) evaluated. The degree of GDM was significantly higher in lesions ≥ CIN 2 compared to lesions ≤ CIN 1 and this methylation pattern was similar in the pre and post fortification groups. These results suggested that the mandatory fortification with folic acid in the United States has not resulted in a change in the degree or the pattern of GDM in cells involved in the process of cervical carcinogenesis.

Comparison Between In Situ Hybridization and Real-time PCR Technique as a Means of Detecting the Integrated Form of Human Papillomavirus 16 in Cervical Neoplasia

Integration of the human papillomavirus (HPV) genome is thought to be one of the causes of cancer progression. However, there is controversy concerning the physical status of HPV 16 in premalignant cervical lesions, and there have been no reports on the concordance between detection of the integrated form of HPV16 by real-time PCR and by in situ hybridization. We investigated specimens of cervical intraepithelial neoplasia (CIN) and invasive carcinomas for the physical status of HPV 16 by real-time PCR and in situ hybridization. The presence of the integrated form was detected by both real-time PCR and in situ hybridization in zero of four cases of CIN1, three of six cases of CIN2, nine of 27 cases of CIN3, and two of six cases of invasive carcinomas. Integrated HPV 16 was present in some premalignant lesions but was not always present in carcinomas. The concordance rate between the two methods for the detection of the presence of the integrated form was 37 of 43 (86%) cases. Real-time PCR and in situ hybridization were found to be complementary and convenient techniques for determining the physical status of the HPV genome. We conclude that a combination of both methods is a more reliable means of assessing the physical status of the HPV genome in cervical neoplasia.