Intraepithelial Squamous Cell Carcinoma Research Articles

Photodynamic therapy in combination with CO2 laser for the treatment of Bowen's disease.

Photodynamic therapy (PDT) involves the activation of a previously administered photosensitizing agent by visible light to induce tumor necrosis. Photosensitizers are topically applied in the treatment of skin tumors to avoid systemic side effects. In this study, we evaluated the feasibility and efficacy of aminolevulinic acid (ALA) as a photosensitizer (ALA-PDT) in combination with CO2 laser in the treatment of Bowen's disease (BD; intraepithelial squamous cell carcinoma). Twenty-two lesions from 18 patients were randomized into two groups: 11 lesions were treated with topical ALA-PDT (180J/cm(2) at 100mW/cm(2)) + CO2 laser for one to three sessions. The remaining 11 lesions were treated with CO2 laser alone, serving as control group. All patients were reviewed at ≤1-week intervals. Biopsies were taken from BD lesions prior to treatment. The initial evaluation was undertaken 1month after treatment, and biopsies were harvested for histological evaluation. Patients who did not respond to the three sessions of treatment were referred to surgical treatment. In the ALA-PDT + CO2 laser group, 72.73% (8/11) of BD lesions showed complete remission, with an overall clearance of 90.91%, and only one recurred (9%) during follow-up. Local side effects included mild erythema, edema, erosion, and burning and/or stinging sensation. No systemic side effects were observed. In the control group, 63.63% (7/11) of lesions had complete remission and the overall clearance was 54.55%. However, five lesions (45.45%) had recurrence. Local side effects included mild to moderate edema, erosion, ulceration, delayed healing, prolonged pain, and scarring. There existed a significant difference in recurrence rate between the two groups (P < 0.05). Moreover, after ALA-PDT plus CO2 laser treatment, complete necrosis was observed in responsive lesions, and 3months later, the atypical BD cells were replaced by normal keratinocytes. Topical ALA-PDT in combination with CO2 laser is safe, effective, and is associated with low recurrence and reduced side effects.

P16INK4A and survivin: Diagnostic and prognostic markers in cervical intraepithelial neoplasia and cervical squamous cell carcinoma

ObjectiveTo explore the the correlations of p16INK4A (p16) and survivin expressions with cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma. MethodsThe p16 and survivin expressions were detected in 50 cervical squamous cell carcinoma tissues, 150 various grades of CIN tissues and 30 normal cervical tissues using immunohistochemistry. All data were analyzed applying SPSS 17.0 software. ResultsThe p16 and survivin expressions showed the presence of statistical significance in cervical cancer, CINI, CINII, CINIII and normal cervical tissues (P<0.05), and the comparison also revealed statistical significance among groups (all P<0.05); the p16 and survivin expressions were positively correlated with the grade of cervical diseases (both P<0.05). Moreover, p16 protein was associated with CIN grade and lymph node metastases in cervical cancer (all P<0.05); survivin protein was also related with clinical stages, CIN grade and lymph node metastases (all P<0.05); the p16 and survivin expressions were positively correlated with cervical cancer (r=0.854, P<0.001), and associated with poor prognosis of cervical cancer. ConclusionBriefly, p16 and survivin expression may be correlated with the clinico-pathological and prognosis of cervical cancer.

Polymorphisms and functional analysis of the intact human papillomavirus16 e2 gene.

High risk human papillomavirus (HR-HPV) E2 proteins play roles in transcriptional regulation and are commonly functionally disrupted when the HPV genome integrates into host chromosomes. Some 15-40% of cancer cases, however, contain an intact E2 gene or episomal HPV. In these cases, polymorphism of the E2 gene might be involved. This study aimed to determine polymorphisms of the E2 gene in episomal HPV16 detected in high grade squamous intraepithelial lesions and squamous cell carcinomas and altered functions compared to the E2 prototype. The E2 gene was amplified and sequenced. Two expression vectors containing E2 gene polymorphisms were constructed and transfected in SiHa and C33A cells, then E6 gene as well as Il- 10 and TNF-α expression was determined by quantitative RT-PCR. Expression vectors and reporter vectors containing the HPV16 long control region (LCR) were co-transfected and transcriptional activity was determined. The results showed that a total of 32 nucleotides and 23 amino acids were changed in all 20 cases of study, found in the transactivation (TA) domain, hinge (H) region and DNA binding (DB) domain with 14, 5 and 13 nucleotide positions. They mostly caused amino acid change. The expressing vectors containing different E2 gene polymorphisms showed E6 mRNA suppression, TNF-α mRNA suppression and IL-10 induction but no statistically significant differences when compared to the E2 prototype. Moreover, promoter activity in HPV16 LCR was not affected by E2 protein with different gene polymorphisms, in contrast to nucleotide variations in LCR that showed an effect on transcription activity. These results demonstrated that E2 gene polymorphisms of episomal HPV16 did not affect transcriptional regulation and suggested that nucleotide variation as well as epigenetic modification of the LCR might play a role in inducing malignant transformation of cells containing episomal HPV16.

Giant pigmented Bowen's disease: A rare variant at a rare site.

Sir, Bowen's disease (BD) is an intraepithelial squamous cell carcinoma (SCC) in situ with a potential for invasive malignancy. Although it has a propensity to occur in sun-exposed areas, other unusual sites are also described.[1] BD is unusual in the pigmented skin and the pigmented variant of BD occurs in less than 2% of cases.[2] Etiology of BD is not clearly understood but various strains of human papilloma viruses, arsenic exposure, and immune suppression have been considered as the risk factors.[3,4] Lesions arising in scars have been described.[5] Three to five percent cases of BD have a risk of developing SCC; metastasis may occur in 1/3 of these invasive tumors.[6] We describe an unusual case of giant pigmented BD over an atypical site. A 68-year-old male presented with a gradually progressing, asymptomatic plaque over the left lower abdomen of 25 years duration. He had applied topical herbal medications with no improvement. There were no medical co-morbidities or excessive sunlight exposure or exposure to arsenic, radiation or underlying immunosuppression. There was history of constant friction at the waistline due to tying of dhoti, a traditional outfit of south India. Cutaneous examination revealed a well defined, hyperpigmented, scaly, verrucous, irregular plaque with a central area of depigmentation measuring about 11 × 5 cm [Figure 1]. Differential diagnoses of Bowen's disease, basal cell carcinoma, and lupus vulgaris were considered. An edge biopsy sent for histopathological examination revealed hyperkeratosis, acanthosis, and irregular elongation of rete ridges with thinned out papillae. Scattered dyskeratotic cells having homogenous eosinophilic cytoplasm and hyperchromatic nuclei were noted imparting a windblown appearance [Figure 2]. Focal areas of dysplasia were noted with a few atypical cells having hyperchromatic nuclei with an intact basement membrane. Scattered melanin pigment was observed within the cytoplasm of the atypical keratinocytes. [Figure 3]. The deeper dermis revealed mild to moderate perivascular chronic inflammatory infiltrate. Multiple sections did not reveal features of SCC. A final diagnosis of pigmented type of Bowen's disease was thus confirmed. Figure 1 Well-defined, hyperpigmented, scaly, verrucous, irregular plaque with a central area of depigmentation measuring about 11 × 5 cm Figure 2 Scattered dyskeratotic cells having homogenous eosinophilic cytoplasm and hyperchromatic nuclei giving a windblown appearance (H and E ×400) Figure 3 Scattered melanin pigment seen within the cytoplasm of the atypical keratinocytes (H and E ×400) In 1912, John T. Bowen described Bowen's disease as a form of in situ SCC, with a potential to progress to invasive carcinoma.[7] Histopathologically, it is characterized by the “windblown” pattern of the epidermis, with loss of polarity and presence of nuclear atypia and mitotic figures.[7] Various treatments including topical imiquimod or 5-fluorouracil, cryotherapy, surgical excision, electrocautery, photodynamic therapy, lasers, and Mohs micrographic surgery have been attempted.[1,2] Excision and primary closure was relatively easy in our patient as a rapid treatment. Cosmetic outcome was not a major consideration for the patient because the lesion was situated in a covered area. Surgical excision was hence chosen ahead of other options such as cryotherapy and topical 5-fluorouracil.[8] BD in sun protected areas of the body is a rare occurrence. It was also a giant lesion (11 × 5 cm) that did not progress to malignancy despite its chronicity. It also occurred at the friction prone site of the waistline due wearing of the dhoti, a traditional garment of south India, hence making it a possible cultural dermatosis. Many waistline dermatoses due to wearing of tight garments such as saree/petticoat have been reported but not Bowen's disease,[9] making our case unique. Financial support and sponsorship Nil. Conflicts of interest There are no conflicts of interest.

Conjunctival Lesions; a Histopathologic Review in Fars Province, Iran.

Purpose:To determine the prevalence of various types of conjunctival lesions, at a tertiary ophthalmic center in Fars Province, South of Iran.Methods:Histopathologic slides and medical records of conjunctival lesions submitted to the pathology department of Khalili Hospital, Shiraz, Iran were reviewed from April, 2009 to July, 2013. The histopathologic diagnoses were categorized into benign, pre-malignant, and malignant lesions. The prevalence of various types of conjunctival lesions was calculated.Results:Histologic sections of 631 conjunctival lesions were reviewed. Benign lesions were most prevalent (81.8%), followed by premalignant (10.8%) and malignant (7.4%) lesions. Pterygia were the most common benign lesions (69.2%), intraepithelial dysplasia constituted most cases of premalignant lesions (94.1%), and squamous cell carcinoma was the most frequent malignant lesion (93.6%). Benign lesions were the most common type of lesions in all age groups, however the prevalence of malignant lesions increased significantly with age (P < 0.001).Conclusion:Benign lesions were the most common conjunctival lesions with pterygia on top of the list, while intraepithelial neoplasia and squamous cell carcinoma were the most common premalignant and malignant lesions, respectively. Conjunctival malignant lesions were more prevalent with older age.

Identification of Human Herpesvirus 8 Sequences in Conjunctiva Intraepithelial Neoplasia and Squamous Cell Carcinoma of Ugandan Patients.

The incidence of squamous cell carcinoma of the conjunctiva is particularly high in sub-Saharan Africa with temporal trends similar to those of Kaposi sarcoma (KS). Human herpesvirus type 8 (HHV8), has not yet been investigated in conjunctiva tumors. In this study biopsies and PBMCs of conjunctiva neoplasia patients along with nonneoplastic conjunctiva tissues have been analyzed for HHV8 sequences by PCR targeting ORF26. All amplimers were subjected to nucleotide sequencing followed by phylogenetic analysis. HHV8 DNA has been identified in 12 out of 48 (25%) HIV-positive, and in 2 out of 24 (8.3%) HIV-negative conjunctiva neoplastic tissues and in 4 out of 33 (12.1%) PBMC samples from conjunctiva neoplasia diseased patients as well as in 4 out of 60 (6.7%) nontumor conjunctiva tissues. The viral load ranged from 1 to 400 copies/105 cells. Phylogenetic analysis showed that the majority of HHV8 ORF26 amplimers clustered with subtypes R (n = 11) and B2 (n = 6). This variant distribution is in agreement with that of HHV8 variants previously identified in Ugandan KS cases. The presence of HHV8 in conjunctiva tumors from HIV-positive patients warrants further studies to test whether HHV8 products released by infected cells may have paracrine effects on the growth of conjunctiva lesions.

Adenocarcinoma in the squamous-lined esophagus without Barrett’s mucosa, probably arising from esophageal gland duct

Endoscopic mucosal resection (EMR) was performed for submucosal tumor (SMT)-like lesion of esophagogastric junction in an 81-year-old male. Histopathological findings revealed the presence of adenocarcinoma with high-grade cytological atypia. No foci of Barrett’s esophagus and/or heterotopic gastric mucosa were detected in the specimen. Intraepithelial squamous cell carcinoma was also not detected in the adjacent stratified squamous layer. The contour of the tumor was similar to normal esophageal gland ducts but tumor did have immunohistochemical staining patterns similar to esophageal cardiac glands. Esophageal adenocarcinoma arising in the background without the foci of Barrett’s esophagus is extremely rare and we reported the case with review of the literature in this study.

Squamous Intraepithelial Lesions and Squamous Cell Carcinoma Detected by Endometrial Sampling: Pap Test Correlation and Outcome Data

Squamous Intraepithelial Lesions and Squamous Cell Carcinoma Detected by Endometrial Sampling: Pap Test Correlation and Outcome Data

MAD2 and CDC20 are upregulated in high-grade squamous intraepithelial lesions and squamous cell carcinomas of the uterine cervix.

Abnormal expression of the spindle assembly checkpoint proteins causes tumor cell aneuploidy, which has been reported in various malignancies. The expression of mitotic arrest deficient 2 (MAD2) and cell-division cycle 20 homolog (CDC20), the key spindle assembly checkpoint proteins, has not been studied in cervical carcinogenesis. In this study, we compared the expression of MAD2 and CDC20 in 332 cases, including normal cervical tissues, low-grade squamous intraepithelial lesions, high-grade squamous intraepithelial lesions (HSILs), and invasive squamous cell carcinomas (SCCs). Both MAD2 and CDC20 were overexpressed in the nuclei or cytoplasm of dysplastic and malignant tumor cells. The frequency of MAD2 overexpression was markedly increased from undetectable (0/100) in normal cervical tissues and 2% (1/50) in low-grade squamous intraepithelial lesions to 67.1% (53/79) in HSILs and 52.4% (54/103) in SCCs. Similarly, CDC20 was overexpressed in 49.4% (39/79) of HSILs and 22.3% (23/103) of SCCs, whereas CDC20 was not detectable (0/100) in normal cervical tissues and overexpressed only in 8.0% (4/50) of low-grade squamous intraepithelial lesions. In SCC cases, MAD2 overexpression correlated with a patient age of less than 60 yr (P=0.043), nonkeratinizing histologic type (P=0.018), and a lesser degree of stromal invasion (P=0.026). In conclusion, MAD2 and CDC20 overexpression was increased in HSILs and SCCs, suggesting their involvement in the initiation of cervical cancers. Controlling CDC20 and MAD2 expression may be a therapeutic strategy for cervical cancer.

MRNA Expression in Cervical Specimens for Determination of Severe Dysplasia or Worse in HPV-16/18–Positive Squamous Lesions

The objective of current study was to determine the p16 mRNA level in cervical cells by relative quantification (RQ) and to test viral E6 expression in human papillomavirus (HPV) -16 or -18-positive specimens by widely used methods. We targeted the pivotal mRNA level associated with severe dysplasia or worse. Cervical specimens were taken from 134 women with cervical disease and 132 women with normal cytologic results. The presence of HPV was analyzed by sequencing. The results of p16 and E6 analyses were statistically processed in receiver operating characteristic curve analysis to predict severe dysplasia or worse. The HPV DNA was detected in 81.4% (109/134) of women with cervical disease and in 27.3% (36/132) of women with normal cytologic results. HPV-16 or -18 were present in 59.7% (80/134) of abnormal specimens. p16 and E6 mRNA expression was increasing with severity of cervical dysplasia. p16 mRNA expression was found 4.35-fold and 13.15-fold increased in high-grade squamous intraepithelial lesions and squamous cell carcinomas, respectively. E6 mRNA expression was significantly increased (p = .0038) in severe dysplasias or worse. The RQ method achieved better sensitivity (82.6%), and E6 mRNA got better specificity (80.6%) for the prediction of severe dysplasia or worse. An increasing level of p16 and E6 mRNA transcripts could mean the potential of cervical dysplasia progression to cancer, but further studies should be done to confirm this proposition. Nevertheless, we consider using both tests to improve the sensitivity and specificity for prediction of severe dysplasia or worse.

Keratin 17 in premalignant and malignant squamous lesions of the cervix: proteomic discovery and immunohistochemical validation as a diagnostic and prognostic biomarker

Most previously described immunohistochemical markers of cervical high-grade squamous intraepithelial lesion (HSIL) and squamous cell carcinoma may help to improve diagnostic accuracy but have a minimal prognostic value. The goals of the current study were to identify and validate novel candidate biomarkers that could potentially improve diagnostic and prognostic accuracy for cervical HSIL and squamous cell carcinoma. Microdissected tissue sections from formalin-fixed paraffin-embedded normal ectocervical squamous mucosa, low-grade squamous intraepithelial lesion (LSIL), HSIL and squamous cell carcinoma sections were analyzed by mass spectrometry-based shotgun proteomics for biomarker discovery. The diagnostic specificity of candidate biomarkers was subsequently evaluated by immunohistochemical analysis of tissue microarrays. Among 1750 proteins identified by proteomic analyses, keratin 4 (KRT4) and keratin 17 (KRT17) showed reciprocal patterns of expression in the spectrum of cases ranging from normal ectocervical squamous mucosa to squamous cell carcinoma. Immunohistochemical studies confirmed that KRT4 expression was significantly decreased in squamous cell carcinoma compared with the other diagnostic categories. By contrast, KRT17 expression was significantly increased in HSIL and squamous cell carcinoma compared with normal ectocervical squamous mucosa and LSIL. KRT17 was also highly expressed in immature squamous metaplasia and in endocervical reserve cells but was generally not detected in mature squamous metaplasia. Furthermore, high levels of KRT17 expression were significantly associated with poor survival of squamous cell carcinoma patients (Hazard ratio=14.76, P=0.01). In summary, both KRT4 and KRT17 expressions are related to the histopathology of the cervical squamous mucosa; KRT17 is highly overexpressed in immature squamous metaplasia, in HSIL, and in squamous cell carcinoma and the level of KRT17 in squamous cell carcinoma may help to identify patients who are at greatest risk for cervical cancer mortality.

A Clinical and Pathological Overview of Vulvar Condyloma Acuminatum, Intraepithelial Neoplasia, and Squamous Cell Carcinoma

Condyloma acuminatum, intraepithelial neoplasia, and squamous cell carcinoma are three relatively frequent vulvar lesions. Condyloma acuminatum is induced by low risk genotypes of human papillomavirus (HPV). Vulvar intraepithelial neoplasia (VIN) and squamous cell carcinoma have different etiopathogenic pathways and are related or not with high risk HPV types. The goal of this paper is to review the main pathological and clinical features of these lesions. A special attention has been paid also to epidemiological data, pathological classification, and clinical implications of these diseases.

Comparison of cervical cytological screening results between postmenopausal and elderly women

There is insufficient data on geriatric period cervical cytology screening in our country. The aim of this study was to compare the cervical cytological results among elderly women and women aged between 45-64 years. A comparison of cervical cytological results of 273 women aged over 65 years and 388 women aged between 45-64 years from January 2010 to June 2013 was performed for presence of cytological abnormalities. Adequacy was assessed according to the criteria of the 2001 Bethesda System. Cervical cytological abnormalities were classified as follows: atypical squamous cell of undetermined significance (ASCUS), cannot exclude a high-grade squamous intraepithelial lesion (ASC-H), low grade squamous intraepithelial lesion (LSIL), high grade squamous intraepithelial lesion (HSIL) and squamous cell carcinoma (SSC). Overall, the prevalence of cytological abnormality was 3% and the prevalence of ASCUS, ASCH, LSIL, and HSIL was 1.8%, 0.3%, 0.6% and 0.3%, respectively. Cytological abnormalities were detected to be higher in the geriatric group in comparison to the postmenopausal group (%5.1 vs. %1.5) (p=0.008). ASCUS was detected to be 3.7% in the geriatric group and 0.5% in the postmenopausal group (p=0.005). There was no statistically significant difference between the groups in means of other cytological anomalies. The reactive inflammatory changes were detected more in the postmenopausal group and atrophic changes in the geriatric group (p < 0.001). It should not be forgotten that although the cervical screening follow-up program is discontinued in elderly women, squamous intraepithelial lesions can still be encountered in this group even if the rate is low.

Association of the Plasma and Tissue Riboflavin Levels with C20orf54 Expression in Cervical Lesions and Its Relationship to HPV16 Infection

Riboflavin deficiency can cause a variety of metabolic problems that lead to skin and mucosal disorders. Limited evidence suggests that high intake of riboflavin may reduce overall risks of cancer. However, association of this deficiency with cervical cancer and precancerous lesions are still not definitively known. In this study, we characterized the relationship between plasma and tissue riboflavin levels and C20orf54 protein expression in patients with cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma (CSCC) as well as the relationship of these levels with human papillomavirus virus 16, 18 (HPV16/18) infections. High-performance liquid chromatography (HPLC) was used to measure blood riboflavin levels in patients with CIN and CSCC, and an enzyme-linked immunosorbent assay (ELISA) was used to determine tissue riboflavin levels in patients with CSCC and matched normal mucous epithelia. The expression of C20orf54 in fresh CSCC and matched tissues were detected by qRT-PCR and western blot, respectively. And it was further confirmed by immunohistochemistry (IHC) with formalin-fixed, paraffin-embedded CIN and CSCC. An HPV genotyping chip was used to analyze HPV infection and typing. The results showed that patients with CIN and CSCC had decreased plasma riboflavin levels as compared with normal controls. There was also significantly decreased riboflavin in tissues from CSCC patients, when compared with normal cervical epithelia. C20orf54 expression were significantly up-regulated in CSCC compared to matched control on both mRNA and protein level. Tissue riboflavin levels were significantly lower in HPV16/18 positive tissue compared with HPV16/18-negative tissue, and an inverse association was found between tissue riboflavin levels and C20orf54 mRNA and protein expression in CSCC. Additionally, C20orf54 was significantly correlated with tumor stages. In conclusion, C20orf54 tend to play a protective role in Uyghur cervical carcinogenesis of which modulating riboflavin absorption, and it is also related with HPV infection.

Clinical management of cervical intraepithelial neoplasia in pregnant and postpartum women

To evaluate the clinical management of cervical intraepithelial neoplasia (CIN) and cervical microinvasive squamous cell carcinoma in pregnant and postpartum women. This prospective study enrolled 27,230 pregnant women undergoing routine gestational examinations between August 1, 2007 and July 31, 2010 in the Beijing Obstetrics and Gynecology Hospital, Capital Medical University. Colposcopy and cervical biopsy were performed for patients with abnormal Thin Prep® Papanicolaou test (TCT) results. Periodic colposcopy was performed every 8-12 weeks and cervical biopsy was performed if progression was suspected. Cervical cold knife conization was recommended to patients diagnosed with CINIII or microinvasive cervical carcinoma 6-12 weeks after delivery. A total of 2,260 patients had abnormal TCT results (8.12 %). Colposcopy and cervical biopsy were performed for 369 patients. Fifteen patients had microinvasive squamous cell carcinoma, 116 patients had cervicitis, and the number of CIN patients with histological grades I, II, and III were 124, 49, and 65, respectively. Tumor progression during pregnancy was found in 253 patients (CINI or above). Prognosis varied depending on the highest grade of pathological diagnosis results during pregnancy or initial pathological diagnosis results performed 6-12 weeks after delivery by cervical biopsy under colposcopy. Treatment and follow-up were carried out according to diagnoses, state of progression, and reversion (if any). These findings underline a need for cervical lesion screening for all women during pregnancy, and colposcopy should be performed for pregnant women who have abnormal TCT results. Appropriate treatment and follow-up were recommended according to different diagnosis of CIN.

Simultaneous cervical screening is required in females diagnosed with anal intraepithelial neoplasia (AIN) or squamous cell carcinoma (SCC)

Simultaneous cervical screening is required in females diagnosed with anal intraepithelial neoplasia (AIN) or squamous cell carcinoma (SCC)

Human papillomaviruses in intraepithelial neoplasia and squamous cell carcinoma of the conjunctiva

The infection with human papillomavirus (HPV) has been described as a risk factor for squamous cell carcinoma of the conjunctiva (SCCC), although the evidence is conflicting. To assess the relation between HPV infection and intraepithelial neoplasia or SCCC, we evaluated archived material from biopsies of the conjunctiva performed at the Maputo Central Hospital (Mozambique) in patients with suspected eye cancer. The quality of DNA was assessed by PCR using β-globin-specific primers. A total of 22 consecutive biopsies (intraepithelial neoplasia, SCCC, and benign conditions) positive for β-globin were further tested for HPV infection by PCR using the general primers GP5+/GP6+ and CPI/CPII. In addition, PCR with type-specific primers HPV 16 and HPV 18 was performed. Nineteen biopsies corresponded to intraepithelial neoplasia (two low-grade and nine high-grade) or SCCC (n=8), from which 11 (57.9%) tested positive for HPV infection; nine were positive for CPI/CPII, including one case also positive for GP5+/GP6+ and HPV 18, and the remaining two tested positive only for HPV 16. HPV DNA was not detected in any of the three biopsies of benign conditions. These results suggest a stronger association between infection with cutaneous HPV and SCCC than for mucosal HPV. However, further research is required to clarify the relation between HPV and SCCC as well as to understand the potential of the HPV vaccine currently available for cervical cancer to prevent SCCC.

Abstract A03: Epigenetic alterations of stratifin in ocular surface squamous neoplasia: Pathogenesis and prognosis

Abstract Purpose: Ocular Surface Squamous neoplasia (OSSN) is the most common tumor of conjunctival epithelium associated with risk of permanent visual impairment. It includes conjunctival intraepithelial neoplasia (CIN) and squamous cell carcinoma (SCC).Of the multifactorial etiopathogenesis of OSSN, ultraviolet irradiation has been reported as a major risk factor in the pathogenesis of OSSN. Sunlight induced p53 mutations are known to contribute towards increased risk of OSSN. Stratifin (14-3-3σ)/HEM (human epithelial marker) is a p53 mediated inhibitor of cell cycle progression and has been shown to be a target of epigenetic deregulation in various carcinomas. In the present study, Stratifin expression, its promoter methylation status and expression of mutant p53 in OSSN was studied. The significance of Stratifin and p53 in the pathogenesis and prognosis of ocular surface squamous neoplasia (OSSN) patients was also evaluated. Methods: Sixty-four histopathologically confirmed OSSN cases (44 SCC and 20 CIN) were included in this study. AJCC TNM staging was performed and patients were followed up for 32 months. Immunohistochemical expression of Stratifin (clone-ab14123) and mutant p53 (clone-DO7) protein was evaluated; methylation status of Stratifin was determined by methylation specific PCR using specific primers. Kaplan–Meier survival and Cox regression analysis was done to assess the prognostic significance of Stratifin and p53. Results: Loss of Stratifin immunoexpression was observed in 75% (48/64) and 48% (31/64) cases showed mutant p53 expression. Hypermethylation of Stratifin promoter was seen in 63% (40/64) OSSN cases. Of the 48 cases with loss of Stratifin expression, promoter hypermethylation was seen in 35% (17/48), mutant p53 expression in 10% (5/48) and both promoter hypermethylation and mutant p53 expression in 58% (23/48) cases. These results indicate that both hypermethylation of Stratifin and mutant p53 expression are associated with loss of Stratifin in OSSN cases. On statistical analysis, Stratifin loss was significantly associated with tumor size &amp;gt;2cm, T3 and T4 category, poor histopathological differentiation of SCC and reduced disease free survival (P 0.05). Cox analysis showed Stratifin to be an independent prognostic marker for OSSN (p =0.03) patients. Mutant p53 expression was associated with history of longer sunlight exposure in 56% cases. No significant correlation was observed between mutant p53 expression and other clinicopathological feature or clinical outcome. Conclusions: Our results indicate that loss of Stratifin is a key event in the pathogenesis of OSSN. Presence of both aberrant methylation of Stratifin and mutant p53 expression implicates involvement of p53-Stratifin mediated signalling pathway in the pathogenesis of OSSN. Stratifin loss may also prove to be a useful biomarker for identifying high risk OSSN patients. Citation Format: Sheetal Chauhan, Seema Sen, Anjana Sharma, Shyam S. Chauhan, Seema Kashyap, Radhika Tandon, Neelam Pushker, M Vanathi. Epigenetic alterations of stratifin in ocular surface squamous neoplasia: Pathogenesis and prognosis. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr A03.

Plasma-free amino acid profiling of cervical cancer and cervical intraepithelial neoplasia patients and its application for early detection

In this study, plasma-free amino acid profiles were used to investigate pre-cancerous cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma (CSCC) metabolic signatures in plasma. Additionally, the diagnostic potential of these profiles was assessed, as well as their ability to provide novel insight into CSCC metabolism and systemic effects. Plasma samples from CIN patients (n = 26), CSCC patients (n = 22), and a control healthy group (n = 35) were analyzed by high-performance liquid chromatography, and their spectral profiles were subjected to the t test for statistical significance. Potential metabolic biomarkers were identified using database comparisons that examine the significance of metabolites. Compared with healthy controls, patients with CIN and CSCC demonstrated lower levels of plasma amino acids; plasma levels of arginine and threonine were increased in CIN patients but were decreased in cervical cancer patients. Additionally, the levels of a larger group of amino acids (aspartate, glutamate, asparagine, serine, glycine, histidine, taurine, tyrosine, valine, methionine, lysine, isoleucine, leucine, and phenylalanine) were gradually reduced from CIN to invasive cancer. These findings suggest that plasma-free amino acid profiling has great potential for improving cancer screening and diagnosis and for understanding disease pathogenesis. Plasma-free amino acid profiles may have the potential be used to determine cancer diagnoses in the early stage from a single blood sample and may enhance our understanding of its mechanisms.

Anal intraepithelial neoplasia and squamous cell carcinoma in HIV‐infected adults

Anal cancer is one of the most common non-AIDS-defining malignancies in the era of combination antiretroviral therapy. Its precursor lesion, anal intraepithelial neoplasia (AIN), is highly prevalent in HIV-infected populations. More than 90% of anal squamous cell cancers are attributable to human papillomavirus (HPV). While the biology of HPV-related intraepithelial neoplasia is consistent across lower anogenital sites, the natural history of AIN is not well established and cannot be assumed to be identical to that of cervical intraepithelial neoplasia. Screening strategies to prevent anal cancer should be developed based on robust natural history data in HIV-infected and uninfected populations. Likewise, treatments need to be tested in randomized clinical trials, and reserved for those at significant risk of progression to cancer. This review covers the epidemiology, pathogenesis and immunology of HPV infection, AIN and anal cancer, and summarizes the current diagnosis, screening and treatment strategies in HIV-infected adults.