Non-keratinizing Squamous Cell Carcinoma Research Articles

Association between high survivin expression and late clinical stage of nasopharyngeal non-keratinizing squamous cell carcinoma

Association between high survivin expression and late clinical stage of nasopharyngeal non-keratinizing squamous cell carcinoma

The EBV-DNA Can be Used as a Diagnostic and Follow-up Parameter of the Rhinopharyngeal Tumors in the Non-Endemic Population of the Western Sicily.

To evaluate whether EBV-DNA can be used as a diagnostic and follow-up parameter for nasopharyngeal tumors in a non-endemic population. The study was carried out in a university hospital. A retrospective study was conducted on 40 paraffin samples of histological preparations. EB-DNA was detected by real-time PCR technique. A prospective study was also conducted on a group of 30 patients who underwent nasopharyngeal biopsy for suspected nasopharyngeal carcinoma (NPC) by comparing EBV-DNA concentrations between the histological specimen and the serum. Quantification of genomic copies of EBV-DNA in serum and detection of anti-EBV antibodies was performed. In both groups the presence of high viral load of EBV-DNA was found in nonkeratinizing squamous cell carcinomas, in three cases of lymphepitomyoma and in 4 out of 6 cases of non-differentiated non-carcinoma lymph node metastases. squamous keratinizing cells. In all cases of NPC, an antibody pattern typical of reactivations (IgGVCA+, IgG-EA+, IgG-EBNA+) and IgA-EA-D, frequently positive in cases of NPC, has been highlighted. A good correlation between the high EBV-DNA charges and the histological diagnosis was highlighted. Our study also found that the assessment of viral EBV load can also be considered in the prognostic evaluation and in the follow-up of patients with NPC.

Cytopathologic features of SMARCB1 (INI-1)-deficient sinonasal carcinoma.

SMARCB1 (INI-1)-deficient sinonasal carcinoma (IDSNC) is a newly described, poorly differentiated carcinoma. Herein, the authors describe the cytopathologic features of IDSNC in fine-needle aspiration (FNA) samples. The pathology archives of 2 academic institutions were searched for cases of IDSNC with available cytologic materials. All available slides were reviewed and a detailed morphological analysis was performed. Six cases were identified from 6 patients. FNA specimens were obtained from metastatic sites (4 cases), a local disease recurrence (1 case), and a primary lesion extending into the orbit (1 case). The majority of cases had cellular specimens with features of nonkeratinizing squamous cell carcinoma composed of cohesive clusters and sheets of oval to polygonal cells with indistinct cell borders present within a background of necrotic debris. Two cases had single rhabdoid cells with more abundant cytoplasm. The nuclei generally were small and uniform with fine chromatin, small nucleoli, and mild nuclear membrane irregularities; moderate anisonucleosis and multinucleation were noted in one case. Apoptotic bodies were common in 5 of 6 cases, and readily identifiable mitoses in 3 of 6 cases. Immunohistochemistry demonstrated that all cases expressed cytokeratins, 4 of 5 cases expressed p63, and all cases demonstrated loss of INI-1 nuclear expression. All cases were negative for neuroendocrine markers and aberrant p16 staining. A diagnosis of IDSNC can be suspected on FNA specimens from patients with high-grade sinonasal carcinomas that have either a nonkeratinizing squamous cell carcinoma or rhabdoid morphology. Clinical history and the availability of material for immunohistochemistry are key in confirming the diagnosis. Cancer Cytopathol 2018. © 2018 American Cancer Society.

Evaluation of matrix metalloproteinase-9 expressions in nasopharyngeal carcinoma patients

Nasopharyngeal carcinoma (NPC) is one of head and neck cancer with a poor prognosis because of the position of the tumor adjacent to the skull base and vital structures. Degradation of extracellular matrix that will cause tumor cells to invade surrounding tissues, vascular or lymphatic vessels. One that plays a role in the extracellular matrix degradation process is matrix metalloproteinase-9 (MMP-9). MMP-9 plays a role in tumor invasion process, metastasis and induction of tumor tissue vascularization. To determine the expression of MMP-9 in patients with nasopharyngeal carcinoma, a descriptive study was conducted by examining immunohistochemistry MMP-9 in 30 NPC tissues that had never received radiotherapy, chemotherapy or combination. Frequency distribution of NPC patient mostly in the age group 41-50 years old and 51-60 years were nine people (30.0%); men (73.3%) and non-keratinizing squamous cell carcinoma (53.3%) histopathology type. The overexpression of MMP-9 in patients with nasopharyngeal carcinoma were mostly found in advance stage.

The interplay of UV and cutaneous papillomavirus infection in skin cancer development.

Cutaneous human papillomaviruses (HPVs) are considered as cofactors for non-melanoma skin cancer (NMSC) development, especially in association with UVB. Extensively studied transgenic mouse models failed to mimic all aspects of virus-host interactions starting from primary infection to the appearance of a tumor. Using the natural model Mastomys coucha, which reflects the human situation in many aspects, we provide the first evidence that only UVB and Mastomys natalensis papillomavirus (MnPV) infection strongly promote NMSC formation. Using UVB exposures that correspond to UV indices of different geographical regions, irradiated animals developed either well-differentiated keratinizing squamous cell carcinomas (SCCs), still supporting productive infections with high viral loads and transcriptional activity, or poorly differentiated non-keratinizing SCCs almost lacking MnPV DNA and in turn, early and late viral transcription. Intriguingly, animals with the latter phenotype, however, still showed strong seropositivity, clearly verifying a preceding MnPV infection. Of note, the mere presence of MnPV could induce γH2AX foci, indicating that viral infection without prior UVB exposure can already perturb genome stability of the host cell. Moreover, as shown both under in vitro and in vivo conditions, MnPV E6/E7 expression also attenuates the excision repair of cyclobutane pyrimidine dimers upon UVB irradiation, suggesting a viral impact on the DNA damage response. While mutations of Ras family members (e.g. Hras, Kras, and Nras) were absent, the majority of SCCs harbored—like in humans—Trp53 mutations especially at two hot-spots in the DNA-binding domain, resulting in a loss of function that favored tumor dedifferentiation, counter-selective for viral maintenance. Such a constellation provides a reasonable explanation for making continuous viral presence dispensable during skin carcinogenesis as observed in patients with NMSC.

Correlation of clinicopathologic features and lung squamous cell carcinoma subtypes according to the 2015 WHO classification

AimsThis study aimed to determine the relationship between clinicopathologic features and lung squamous cell carcinoma (LSCC) subtypes according to the 2015 WHO classification. MethodsWe identified 824 operable LSCC patients undergoing a complete surgical resection at Shanghai Chest Hospital between April 2015 and January 2017. Immunohistochemistry was used to investigate the clinicopathologic features. ResultsAmong them, the percentages of LSCC subtypes were 66.1% (545/824), 28.6% (236/824), and 5.2% (43/824) for keratinizing squamous cell carcinoma (KSCC), nonkeratinizing squamous cell carcinoma (NKSCC), and basaloid squamous cell carcinoma (BSCC), respectively. There were more males, more smokers, and more pneumonectomy surgeries in KSCC patients (p = 0.008, p = 0.000, p = 0.043). There were more N2 lymph node involvement and pathological stage III in NKSCC patients (p = 0.01, p = 0.03). BSCC did not demonstrate specificity to anything, but expressed adenocarcinoma markers more frequently. No significant difference existed between pathological subtypes and other clinicopathologic features, such as age, location type, visceral pleural involvement and lymphovascular invasion. The frequencies of EGFR sensitive mutations and ALK rearrangements were not significantly different among three subtypes. ConclusionSignificant relationships exist between some clinicopathologic features and LSCC subtypes.

Refining the Role of Lymph Node Biopsy in Survival for Patients with Nasopharyngeal Carcinoma: Population-Based Study from the Surveillance Epidemiology and End-Results Registry.

The updated version of the National Comprehensive Cancer Network (NCCN) guidelines revised pretreatment workup for nasopharyngeal carcinoma (NPC) into "biopsy of the primary site or neck." Despite provision of important diagnostic information, concerns regarding tumor cell dissemination limit the application of lymph node biopsy. This study aimed to investigate whether biopsy of the neck is associated with impaired survival in NPC. A propensity score-matched, population-based cohort identified from the Surveillance, Epidemiology, and End Results database was used to compare overall survival (OS) and disease-specific survival (DSS) of patients who underwent pretreatment cervical lymph node biopsy without subsequent neck dissection or removal of node compared with patients who did not undergo node biopsy. Of 2910 eligible patients, 416 (14.3%) underwent pretreatment lymph node biopsy. After use of control for patient, tumor, and demographic characteristics, biopsy was not associated with impaired OS (hazard ratio [HR], 1.15; 95% confidence interval [CI] 0.89-1.47; P=0.29) or DSS (HR, 1.07; 95% CI 0.81-1.40; P=0.63). Interestingly, in the subgroup analysis, the unfavorable effect of biopsy was observed for patients with differentiated non-keratinizing squamous cell carcinoma (but not other histologic types). Race did not positively alter the survival outcomes. The findings provide reference for clinical practice, showing that pretreatment cervical lymph node biopsy is not associated with impaired survival in NPC, except for patients with differentiated non-keratinizing squamous cell carcinoma. The recommended NCCN guidelines would be more specific by adding details to the general recommendation that neck biopsy is safe for all patients. Future prospective studies are needed to verify the study findings.

Abstract 5759: Histopathologic patterns of nasopharyngeal malignancy in Lagos, Nigeria and its association with Epstein-Barr virus (EBV): A 10-year retrospective study

Abstract Background: Nasopharyngeal cancer (NPC) is a rare cancer worldwide. It is now found to be increasing in incidence in Nigeria, though the incidence is lower when compared to countries in the Mediterranean basin, North Africa and Southeast Asia. NPC is frequently associated with Epstein-Barr virus (EBV), but no previous study has been done in Lagos to document the association. Objective: To determine the prevalence, histologic types and age distribution of nasopharyngeal malignancy in Lagos University Teaching Hospital (LUTH) and its association with EBV over a period of 10 years, between January 2003 to December 2012. Materials and Methods: Slides and formalin-fixed paraffin-embedded (FFPE) tissue blocks, hospital request forms, referral cards, patients’ case notes and duplicate copies of histopathology reports of cases of histologically diagnosed nasopharyngeal malignancy in the Anatomic and Molecular Pathology department of LUTH within the study period were retrieved. The cases were reclassified into epithelial and non-epithelial malignancies. The epithelial malignancies were reclassified according to WHO classification. Immunohistochemistry (IHC) for EBV latent membrane protein-1 (LMP-1) was carried out on cases with viable FFPE to evaluate the prevalence of the virus in Nasopharngeal malignancy. Results: A total of 75 histologically confirmed cases of nasopharyngeal malignancy were seen during the study period, which constituted 1.1% of the total cancer recorded in this center. There were 47 males (63%) and 28 females (37%) with a male to female ratio of 1.7:1. The age ranged from 3 to 75 years, mean age was 44.1 years, the peak age group was 40 -49 years for males and 50 - 59 years for females. On histological classification, undifferentiated carcinoma (WHO type III) was the commonest (49.3%), differentiated non-keratinizing squamous cell carcinoma, SCC (WHO type II) and keratinizing SCC (WHO type I) accounted for 24% and 9.3% respectively. Thus, NPC accounted for 82.6% of the total cases of nasopharyngeal malignancy seen. Other histological types seen were non-Hodgkin lymphoma (NHL) 13.3%, well differentiated adenocarcinoma 2.7% and low grade mucoepidermoid carcinoma 1.3%. EBV LMP-1 IHC was positive in 30 (86%) out of the 35 cases studied. These consisted of 14 undifferentiated carcinoma, 5 NHL, 2 adenocarcinoma, 7differentiated non-keratinizing SCC (WHO type II) and the 2 cases of keratinizing SCC (WHO type 1). Conclusion: Nasopharyngeal malignancy is not uncommon in this environment. It occurred more frequently in males than females with the peak age of incidence at 5th decade of life, undifferentiated carcinoma (WHO type III) is the commonest histologic type and it is highly associated with EBV. Note: This abstract was not presented at the meeting. Citation Format: Oladipo Omoseebi, Olakanmi Akinde, Omobolade Obadofin, Nzechukwu Ikeri, Gabriel Ogun, Adekumbiola Banjo. Histopathologic patterns of nasopharyngeal malignancy in Lagos, Nigeria and its association with Epstein-Barr virus (EBV): A 10-year retrospective study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5759. doi:10.1158/1538-7445.AM2017-5759

Squamous and Neuroendocrine Specific Immunohistochemical Markers in Head and Neck Squamous Cell Carcinoma: A Tissue Microarray Study.

The performance characteristics of neuroendocrine-specific and squamous-specific immunohistochemical markers in head and neck squamous cell carcinomas (SCC), in particular in oropharyngeal tumors in this era of human papillomavirus(HPV)-induced cases, are not well-established. The differential diagnosis for poorly differentiated SCCs, for nonkeratinizing oropharyngeal SCCs, and for other specific SCC variants such as basaloid SCC and undifferentiated (or lymphoepithelial-like) carcinomas includes neuroendocrine carcinomas. Given that neuroendocrine carcinomas of the head and neck are aggressive regardless of HPV status, separating them from SCC is critically important. In this study, we examined the neuroendocrine markers CD56, synaptophysin, and chromogranin-A along with the squamous markers p40 and cytokeratin 5/6 in a large tissue microarray cohort of oral, oropharyngeal, laryngeal, and hypopharyngeal SCCs with known HPV results by RNA in situ hybridization for the oropharyngeal tumors. Results were stratified by site and specific SCC variant. The neuroendocrine stains were rarely expressed in SCC (<1% overall) with CD56 the least, and chromogranin-A the most, specific markers. Further, p40 and cytokeratin 5/6 were very consistently expressed in all head and neck SCC (>98% overall), including very strong, consistent staining in oropharyngeal HPV-related nonkeratinizing SCC. Undifferentiated (or lymphoepithelial-like) carcinomas of the oropharynx are more frequently p40 or cytokeratin 5/6 negative or show only weak or focal expression. In summary, markers of neuroendocrine and squamous differentiation show very high specificity and sensitivity, respectively, across the different types of head and neck SCC.

Clinicopathologic features and genetic profile of the redefined large cell lung carcinoma

Objective: To investigate the clinicopathologic features and genetic profile of large cell lung carcinoma (LCC) redefined by new classification. Methods: Basing on 2015 WHO classification criteria in redefining large cell lung carcinoma, the expression of specific markers (TTF1, Napsin A, p40, CK5/6, CK, vimentin and ZEB1) was detected by immunohistochemistry and D-PAS staining in 303 surgically-removed lung specimens previously diagnosed as large cell lung carcinoma. The clinicopathologic and genetic characteristics (including EGFR, KRAS, BRAF, ALK and ROS1 gene mutation) were analyzed. Results: Based on the new definition of LCC, 116 cases (116/303, 38.3%) of LCC formerly diagnosed were reclassified as solid adenocarcinoma, 49 cases (49/303, 16.2%) as squamous cell carcinoma, 6 cases (6/303, 2.0%) as adenosquamous carcinoma, 22 cases (22/303, 7.3%) as spindle cell carcinoma and only 110 cases (110/303, 36.3%) as large cell carcinoma. Redefined LCCs were characterized as middle-age (range 40-80), male (102/110, 92.7%) and smoking patients (64/110, 58.2%) with intermediate-advanced stage. Among 110 cases, 9 cases with EGFR mutation and 10 cases with KRAS mutation and 1 case with ALK fusion were found. No BRAF and ROS1 alterations were identified. Conclusions: According to the new classification, LCCs formerly diagnosed are mostly reclassified as adenocarcinoma and non-keratinizing squamous cell carcinoma. The newly defined LCC may significantly benefit from clinical therapy.

An immunohistochemical analysis of lymphocytic infiltrations in canine skin cancers.

Lymphocytic infiltrations located in the extracellular matrix often accompany canine skin cancer. They can be characterised as an inflammatory infiltration and/or a second tumour - lymphoma. The aim of this study was an immunohistochemical analysis of a lymphocytic infiltration which accompanies spontaneous skin cancer. Twenty basal cell carcinoma, 20 non-keratinizing squamous cell carcinoma, 20 keratinizing squamous cell carcinoma and 8 sebaceous gland carcinoma samples which were accompanied by a lymphocytic infiltration and/or secondary lymphatic follicles were verified histopathologically. The expression of bcl-2, CD3, CD79α, Ki-67, MCM-3 and MCM-7 in the lymphocytic infiltration was evaluated. Four types of lymphocytic infiltrations were found: I - diffuse bcl-2+, II - diffuse bcl-2-, III - follicular bcl-2+/- where the centre was bcl-2-, and the marginal zone of the follicles and the extrafollicular area were bcl-2+ and IV - aggregated bcl-2+, where the centre and periphery were bcl-2+. The I and IV type corresponds to lymphoma, II type is non-neoplastic immune response and III type suggest reactive follicular hyperplasia. The proliferation of lymphocytes which demonstrated the expression of neoplastic markers (I and IV), suggests preneoplastic phase (pseudolymphoma) or lymphoma - the second independent tumour. A high proliferative index of the follicular blc-2+/- follicular infiltration indicates an increased immunological response of the host against skin cancer.

Transcriptionally Active High-Risk Human Papillomavirus is Not a Common Etiologic Agent in the Malignant Transformation of Inverted Schneiderian Papillomas.

The role of human papillomavirus (HPV) as an etiologic and transformational agent in inverted Schneiderian papilloma (ISP) is unclear. Indeed, reported detection rates of HPV in ISPs range from 0 to 100%. The true incidence has been confounded by a tendency to conflate high- and low-risk HPV types and by the inability to discern biologically relevant from irrelevant HPV infections. The recent development of RNA in situ hybridization for high-risk HPV E6/E7 mRNA now allows the direct visualization of transcriptionally active high-risk HPV in ISP, providing an opportunity to more definitively assess its role in the development and progression of ISPs. We performed p16 immunohistochemistry and high-risk HPV RNA in situ hybridization on 30 benign ISPs, 7 ISPs with dysplasia, 16 ISPs with carcinomatous transformation, and 7 non-keratinizing squamous cell carcinomas (SCCs) with inverted growth that were unassociated with ISP. Transcriptionally active HPV was not detected in any of the 52 ISPs including those that had undergone carcinomatous transformation, but it was detected in two of seven (29%) non-keratinizing SCCs that showed inverted growth. There was a strong correlation between high-risk HPV RNA in situ hybridization and p16 immunohistochemistry (97%; p < 0.01). These results indicate that transcriptionally active high-risk HPV does not play a common role in either the development of ISP or in its transformation into carcinoma.

Lymphoepithelioma is a nonkeratinizing squamous cell carcinoma with Epstein-Barr virus infection in China.

The objective of this study is to investigate the histogenesis of lymphoepithelial carcinoma (LEC) and its relationship with Epstein-Barr virus (EBV). The expression of EBV was detected using in situ hybridization, and the CK5/6, p63, and p40 expression levels were detected using immunohistochemistry in 45 paraffin-embedded tissues from LEC. In 45 paraffin-embedded LEC tissues from 10 different samples, the positive CK5/6 signals were located in the cell membrane. The positive signals for p63 and p40 were located in the cell nucleus. In all LEC cases, the positive rates of CK5/6, p63, and p40 were 93.3% (42/45), 95.6% (43/45), and 93.3% (42/45), respectively. The positive EBV-encoded RNA (EBER) signals were located in the cell nucleus. In the 45 LEC cases, the expression of EBER was strongly positive with a positive rate of 100% (45/45). LEC is closely related to EBV, and EBV plays an important role in the development of LEC. LEC showed positive squamous cell markers, indicating that the samples contain squamous cell carcinoma (SQCC). LEC is EBV (+) with nonkeratinizing SQCC, and this name better reflects the nature of this disease.

MTE11.01 The Clinical Impact of the 2015 WHO Classification of Lung Tumors

MTE11.01 The Clinical Impact of the 2015 WHO Classification of Lung Tumors

Histological Phenotype of Breast Cancer in Indigenous Malawian Women Presenting at Queen Elizabeth Central Hospital in Malawi

Background: Breast cancer is the most commonly diagnosed cancer in women and the second leading cause of death of women in Africa. Management of breast cancer depends on availability of robust clinical and pathologic prognostic and predictive factors to guide decision making and the choice of treatment options. Objective: We set out to determine the common histological phenotypes of breast cancer in indigenous Malawian women presenting at Queen Elizabeth Central Hospital (QECH). Methods: We analyzed 63 samples of breast cancer Formalinfixed paraffin-embedded (FFPE) tissues and determined the prevalence of breast cancer over the three year period. Histological phenotypes were matched for age, tumor grade and clinical signs at presentation. The pathology files were retrieved from January 2012 through December 2013 and January through December 2015. Sixty-three breast cancer cases were analyzed for our results showed an increase in the prevalence of breast cancer from 2012 to 2015 as follows 2012 (22/99) 22.2%, 2013 (25/66) 37.9% and 2015 (16/48) 33.3%. Data was analyzed using IBM SPSS® version 20.0 for windows. The chi-square statistical test was used and a p-value of equal to or less than 0.05 was used to indicate statistical significance of the results Results: The histological phenotypes were categorized as follows, invasive ductal carcinoma (IDC) 26 (41.3%), lobular carcinoma 17 (27%), Lobular plus ductal 4 (6.3%), medullary carcinoma 5(7.9%) and papillary intra ductal carcinoma 3(4.8%). Rare breast cancers were 1.6% each with the following proportions, clear cell carcinoma1, Paget disease and squamous cell carcinoma of surface origin1, invasive micro papillary carcinoma 1, oncocytic carcinoma1, solitary myeloma 1, malignant mesenchymal tumor 1, nonkeratinizing squamous cell carcinoma of surface origin plus Ductal carcinoma in situ 1(DCIS) and signet ring variety of mucinous carcinoma1. The common histological grade were grade 2, 32 (50.8%), grade 1,22 (34.9%) and grade 3,4 (6.4%), 5 (7.9%) cases could not be graded Conclusion: The commonest histological phenotype of breast cancer was invasive ductal carcinoma (IDC) followed by lobular carcinoma, and grade two was the commonest histological grade followed by grade one. Most affected age groups were below 50 years of age with a peak at 41 to 50 years of age

肺原発リンパ上皮腫様癌と鑑別を要した非角化型扁平上皮癌の1例

背景．リンパ上皮腫様癌（lymphoepithelioma-like carcinoma：LELC）は，上咽頭癌の一種であるリンパ上皮腫と類似した組織形態を呈する非常に稀な腫瘍である．症例．65歳男性．CTで左下葉に結節影を指摘され，増大傾向が認められたため，開胸生検の方針となった．術中迅速診断で低分化癌と診断され，左下葉切除術を施行した．病理組織学的には，LELCと矛盾しない所見であった．WHO分類第4版では，LELCと診断するためにはEpstein-Barrウイルス（EBV）の関与が証明される必要がある．本症例ではin situ hybridizationや免疫染色でEBV感染が証明されなかったため，炎症細胞浸潤を伴う非角化型扁平上皮癌と診断された．術後1年3ヶ月経過し無再発生存中である．結論．Hematoxylin-eosin（HE）染色でLELCの所見を有するが，EBV陰性の症例を経験した．LELCは低分化癌に比べ予後良好であり，慎重に診断するべきであると考えられる．EBV陰性でLELCの組織形態を呈する症例をどのように扱っていくかが課題である．

Association of Epstein–Barr Virus (EBV) with malignancy of the nasopharynx in Lagos, Nigeria

Background: Nasopharyngeal cancer is a rare cancer worldwide. It is now found to be increasing in incidence in Nigeria, though the incidence is lower when compared to countries in the Mediterranean basin, North Africa and Southeast Asia. This cancer is frequently associated with Epstein-Barr virus (EBV), but no previous study was done in this centre to document the association. Aim and Objectives: To assess the EBV status in Nigerian patients with nasopharyngeal cancer by using Latent membrane protein – 1 immunohistochemistry. ( LMP-1IHC). Materials and Methods: Thirty-five (35) out of Seventy-five (75) histologically-confirmed cases of nasopharyngeal malignancy (NPM) seen in the Department of Anatomic and Molecular Pathology of Lagos University Teaching Hospital, Lagos between January 2003 and December 2012 were available for EBV study using LMP-1 IHC. Results: EBV LMP-1 IHC was positive in thirty (86%) out of the thirty-five cases studied and these consisted of, 14 cases of undifferentiated carcinoma (WHO type 3), 5 cases of non-Hodgkin lymphoma and 2 cases of adenocarcinoma. Also seven out of eleven cases of differentiated non-keratinizing squamous cell carcinoma (WHO type 2) were EBV positive and two of the three cases of keratinizing squamous cell carcinoma (WHO type 1) were EBV positive. Conclusion: This study has shown that nasopharyngeal malignancy in our centre is highly associated with EBV latency as detected by LMP-1 immunohistochemistry.

PUB094 Correlation of Clinicopathologic Features and Lung Squamous Cell Carcinoma Subtypes According to the 2015 WHO Classification

This study aimed to determine the relationship between clinicopathologic features and lung squamous cell carcinoma subtypes according to the 2015 WHO classification. We identified 368 operable lung squamous cell carcinoma patients who had undergone a complete surgical resection at Shanghai Chest Hospital between April 2015 and March 2016. Among all patients, the percentages of lung squamous cell carcinoma subtypes were: 68.5% (252/368), 26.2% (97/368), and 5.1% (19/368) for keratinizing squamous cell carcinoma (KSCC), nonkeratinizing squamous cell carcinoma (NKSCC), and basaloid squamous cell carcinoma (BSCC), respectively. There were more smokers in patients with KSCC than in patients with other subtypes (p=0.004). There were no significant relationships between pathological subtypes and other clinicopathologic features, such as gender, age, location type, type of resection, stage, visceral pleural involvement, lymphovascular invasion, and lymph node involvement. The expressions of P40, CK5/6, TTF-1, Napsin A, and CK7 were also not significantly different in three subtypes. Our study revealed that there were no significant relationships between clinicopathologic features and lung squamous cell carcinoma subtypes.

An unexpected reason for elevated human chorionic gonadotropin in a young woman Cervical squamous carcinoma

Human chorionic gonadotropin has been used for decades, in addition to specific investigations, to detect pregnancy, trophoblastic tumors, as well as congenital defects. Rarely, it can be elevated in non-trophoblastic tumors such as squamous cell cancers and germ cell tumors. A 33-year-old Asian Syrian female had irregular menses accompanied with feelings of heaviness in the vagina. In addition to routine investigations, we measured the serum beta human chorionic gonadotropin (ß-HCG) level (based on the patient’s complaint of amenorrhea), which was 50.05 ml UI/ml. Cervical biopsy revealed a non-keratinized large cell squamous carcinoma. After excluding other causes, ß-hCG elevation was explained by the ectopic secretion of cancer cells line. Cervical biopsy was suggestive of large cell non-keratinizing squamous cell carcinoma and positive for human chorionic gonadotropin on immunohistochemistry. As a result, we manage the possibility of ectopic secretion of ß-HCG from non- trophoblastic disease.

Abstract 3347: Establishment of four novel HPV-16 positive cell lines of invasive cervical carcinoma of Indian origin and characterization of CD133 positive cancer stem cells

Abstract Background and Aim: Cancer of the cervix is the 2nd most common cancer in Indian women. Cell lines are essential in vitro models in cancer research. Till date, available cell lines in cervical cancer were highly passaged, developed from Caucasian, Black or Asian origin; hence, establishment of a low-passage cervical cancer derived cell line was the first aim of this study. Cancer stem cells (CSCs) are proposed to be responsible for metastasis and therapy resistance. Previously, we reported CD133 to be a putative marker for CSC, and they were further characterized. Materials and Methods: 25 women were recruited after informed consent and approval from the Institute Ethics committee. A small portion of the biopsy was collected in DMEM/F12 media and subjected to primary culture. Successful propagation was possible in 4 cases. Their morphology, epithelial nature and HPV status were evaluated. Variable Number Tandem Repeat (VNTR) assay, cell cycle and kinetic studies were performed. Tissue expression of CD133 and CD49f stem cell markers was evaluated by direct immunofluorescence on frozen tumor samples. Adherent cells were sorted into CD133+ vs CD133- tumor bulk from all 4 cell lines by FACS. These two populations were evaluated for differences in tumorosphere formation, chemosensitivity to cisplatinum, expression of stemness (SOX2, OCT4, NANOG) and EMT (SNAIL, SLUG, TWIST, VIMENTIN and E-CADHERIN) markers at the transcript level. Observations: Four cell lines designated RSBS-9, RSBS-14, RSBS-23 derived from non-keratinizing squamous cell carcinoma and RSBS-43 derived from adenocarcinoma cervix were established and passaged up to 50 times. The epithelial nature of cell lines was confirmed by cytokeratin and epithelial membrane antigen positivity. VNTR assay confirmed derivation of cell lines from respective parental tissue sample. All 4 cell lines were HPV-16positive. The cell doubling time was approximately 48 hours. Immunofluorescence on tissue samples revealed diffuse positivity for CD49f indicating that it may not represent a CSC marker as previously reported whereas CD133 staining showed scattered positive cells. No particular location of CD133+ CSC was observed. Comparison of CD133+ with CD133- bulk population cells revealed increased tumorosphere formation; however, no significant difference in the chemosensitivity to cisplatinum was seen. Real time quantitation of transcripts of stemness and EMT markers revealed CD133+ cells showing upregulation of EMT markers VIMENTIN, SNAIL, SLUG in the RSBS-23 cell line and of VIMENTIN and TWIST in RSBS-9 cell line whereas there was no significant difference in the RSBS-14 and RSBS-43 cell lines for any of the markers studied. Conclusion: Low passage cell lines are useful model systems for understanding the biology of cervical cancer. CD133+ Cancer Stem cells exhibit some EMT markers in 2 of the 4 cell lines established. Citation Format: Shifa Javed, Bal Krishan Sharma, Sanjeev Kumar Sharma, Swati Sood, Rashmi Bagga, Shalmoli Bhattacharyya, Radhika Srinivasan. Establishment of four novel HPV-16 positive cell lines of invasive cervical carcinoma of Indian origin and characterization of CD133 positive cancer stem cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3347.