Aberrant Expression Of Cytokeratin Research Articles

Novel Ratiometric Electrochemical Biosensor for Determination of Cytokeratin 19 Fragment Antigen 21-1 (Cyfra-21-1) as a Lung Cancer Biomarker

Previous studies confirm that aberrant expression of cytokeratin 19 fragment antigen 21-1 (cyfra-21-1) is highly correlated with non-small cell lung cancer (NSCLC). Herein, a proportional electrochemical immunosensor based upon a methylene blue/cadmium telluride/molybdic sulfide (MB/CdTe/MoS2) nanocomposite was successfully fabricated for detecting tumor marker cyfra-21-1. The result indicates that MB/CdTe/MoS2 modified on the surface of a glassy carbon electrode (GCE) supplies a large surface area for immobilizing primary antibodies (Ab1), ensuring high selectivity and sensitivity. To further amplify the sensitivity of the biosensor, ferrocenecarboxylic acid (Fc-COOH) linked with amino-functionalized secondary antibodies through amido bond (Ab2-Fc) as the “signal-on” probe and MB-modified with MoS2 with a large specific surface area as the “signal-off” probe increase the number of active sites for integration of Ab2-Fc/Ag/BSA/Ab1/MB/CdTe/MoS2/GCE electrode with the linker cyfra-21-1 (Ag). Meanwhile, CdTe accelerates the electron transfer and enhances the MB current, improving the sensitivity and detection limit. The presence of cyfra-21-1 led to both the decreasing response of MB and increasing current responses of Fc upon the electrode. Based upon this strategy, the determination of cyfra-21-1 was achieved in a linear range from 10 pg/mL to 10,000 ng/mL with the lowest detectable concentration of 10 pg/mL (signal-to-noise ratio of 3). The electrochemical immunosensor performs a sensitive determination of cyfra-21-1, possibly providing a practical strategy for the clinical analysis of lung cancer.

Aberrant expression of cytokeratin in plasma cell neoplasms: A pitfall for diagnostic errors.

Aberrant expression of cytokeratin in plasma cell neoplasms: A pitfall for diagnostic errors.

Rhabdomyosarcoma Presenting as Bilateral Cervical Lymphadenopathy in an Adult with Aberrant Expression of Cytokeratin and Synaptophysin: A Potentially Serious Diagnostic Pitfall

Rhabdomyosarcoma (RMS) is the most common soft tissue malignancy in children and adolescents. Its occurrence in patients more than 18 years of age is rare. Subtypes of rhabdomyosarcoma include embryonal RMS, alveolar RMS, pleomorphic RMS and spindle/sclerosing RMS.Herein, we report the case of a 42-year-old female patient who presented with bilateral cervical lymphadenopathy of two-month duration. Clinical and radiological examination showed no other significant lesions. Fine needle aspiration cytology and biopsy of lymph node showed diffusely arranged atypical round cells. The clinical findings and morphology pointed to a hematolymphoid malignancy. On immunohistochemical examination, the atypical cells were negative for CD45, CD20, CD3, PAX5 and CD30. The cells showed patchy moderate positivity for cytokeratin, focal weak positivity for synaptophysin and were negative for chromogranin, CD56 and TTF1. On further evaluation, patchy strong positivity for desmin, myogenin and myo D1 were noted. Correlating the morphology and immunoprofile, diagnosis of rhabdomyosarcoma was given.Multiple peripheral lymph node enlargement without an obvious primary lesion is a rare presentation of alveolar RMS, with only a few cases reported in literature. Unusual clinical presentation complicated with aberrant expression of epithelial markers may make the diagnosis of alveolar RMS difficult. This case demonstrates the importance of considering extensive differential diagnosis for neck masses with poorly differentiated morphology.

Petrous apex non-secretory plasma cell myeloma with aberrant cytokeratin expression

ABSTRACTMultiple myeloma is a monoclonal proliferation of plasma cells with common involvement of vertebrae, ribs and skull vault. However, involvement of skull base is relatively uncommon and myeloma manifesting initially as a petrous apex mass is distinctly rare. We report a rare case of non-secretory multiple myeloma in a 52-year-old Egyptian male presenting primarily as a right petrous apex mass with abducens nerve palsy. Additionally, neoplastic cells aberrantly expressed cytokeratin. Although rare, plasma cell myeloma should be considered in the differential diagnosis of petrous apex masses.

A Rare Case of Plasmablastic Lymphoma with Gastric Involvement

Plasmablastic lymphoma (PBL) is a relatively rare, but highly aggressive lymphoma commonly found in the setting of HIV disease.PBL is defined by the World Health Organization as a “diffuse proliferation of large neoplastic cells most of which resemble B-cell immunoblasts, but in which all tumor cells have a plasma cell immunophenotype.”Here we present a unique case of Ann Arbor Stage IV PBL with extranodal gastric involvement diagnosed in an immunocompetent male. An 80-year-old Hispanic male presented to the emergency department with nausea and epigastric pain for four days duration. The patient also endorsed progressive dysphagia to solids and liquids over the past several weeks prior to presentation. Significant findings on physical exam included a pale, well-nourished male with mild epigastric tenderness on light palpation. CT scan revealed diffuse thickening of the gastric wall with a 7.4cm x 6.5cm intraluminal infiltrating mass with luminal narrowing. An EGD was subsequently performed for better visualization and biopsy (Figure 1). Histopathologic studies ultimately identified the mass as malignant plasmablastic lymphoma with aberrant cytokeratin expression. HIV testing was negative but EBV IgG antibody testing was abnormally high. Immunophenotype analysis revealed that CD-138 was strongly positive, CD-20 was negative, CD-45 was weakly positive, and Ki-67 expression was greater than 90% (Figure 2). The patient was started on CHOP chemotherapy regimen.2659_A Figure 1. Large, 5cm, semi-circumferential, easily friable and fungating mass as observed from GE junction during EGD2659_B Figure 2. Histology and Immunophenotype Analysis. (A) High-power magnification showing atypical plasma cells. (B) CD-20 negative. (C) CD-138 positive. (D) Ki-67 Index > 90%.Recent meta-analysis has shown that the median age of diagnosis for HIV-negative PBL is 55 years. Although the pathogenesis of PBL remains incompletely understood, the association with EBV infection and the virus' role in prevention of apoptosis of B cells is well documented.EBV positivity and CD45 expression represent good prognostic indicators for this patient; on the other hand, Ann Arbor Stage IV classification and Ki-67 expression > 80% are typically associated with poor outcome. After the oral mucosa, the GI tract is the most common site of involvement for HIV-negative PBL cases. This case represents a rare phenotypic variant in which the BCL-6 transcription factor is positive, suggesting that perhaps this case is a hybrid of diffuse large B-cell lymphoma (DLBCL) and PBL. Despite the recent advances in the treatment for PBL, prognosis remains poor with the median overall survival of nine months and a 2-year overall survival rate of 10% for HIV-negative PBL.

CK7/CK19 index: A potential prognostic factor for postoperative intrahepatic cholangiocarcinoma patients.

Frequently aberrant expression of cytokeratin 7 (CK7) and cytokeratin 19 (CK19) have been observed in several human cancers. In this retrospective study, we aimed at investigating the prognostic significance of CK7 and CK19 in intrahepatic cholangiocarcinoma (ICC). Immunohistochemistry was performed to assess CK7 and CK19 expression on tissue microarrays in training cohort enrolling 214 ICC patients and validation cohort comprising 108 ICC patients. Kaplan-Meier analysis, Cox's proportional hazards regression, and nomogram were applied to evaluate the prognostic significance of both CKs. Both CK7 and CK19 expression were significantly up-regulated in ICC compared to their non-tumor counterparts, and positively correlated with aggressive tumor phenotypes, like lymph node metastasis and larger tumor size. Furthermore, high expression of either CK7 or CK19 predicted a significantly dismal postoperative survival. Integrated analysis of CK7 and CK19 expression was identified as a better indicator for survival probability. Notably, the nomogram integrating CK7/CK19 index had a perfect prognostic performance as compared with current staging systems. The results were further confirmed in the validation cohort. CK7/CK19 index was an independent adverse prognostic factor for ICC patients' survival, and may be helpful to improve postoperative risk stratification and individualized treatment strategies.

A sandwich-type electrochemical immunoassay for ultrasensitive detection of non-small cell lung cancer biomarker CYFRA21-1

Many studies confirm that the aberrant expression of Cytokeratin 19 fragment 21-1 (CYFRA21-1) is highly correlated with non-small cell lung cancer (NSCLC), especially for squamous cell carcinoma. Herein, we report a sandwich-type electrochemical immunosensor based on signal amplification strategy of multiple nanocomposites to test CYFRA21-1 selectively and sensitively. The proposed immunosensor fabricated by three-dimensional graphene (3D–G), chitosan (CS) and glutaraldehyde (GA) composite on the glass carbon electrode (GCE) with a large surface area is prepared to immobilize primary antibodies (Ab1) and provide excellent conductivity. To further amplify the electrochemical signal, the trace tag on the foundation of gold nanoparticles (AuNPs) is coated with amino-functionalized carbon nanotube (MWCNT-NH2) nanocomposite through thionine linking, which provides more amino groups to capture more horseradish peroxidase-labeled antibodies (HPR-Ab2) and enhances the conductivity. Under optimal conditions, the developed immunosensor exhibits excellent analytical performance for the determination of CYFRA21-1 with a wide linear range from 0.1 to 150ng·mL−1 and a low detection limit (LOD) of 43pg·mL−1. Furthermore, satisfactory results are obtained for the determination of CYFRA21-1 in real clinical serum samples, indicating the potential of the immunoassay to be applied in clinical analysis.

The Aberrant Expression of Cytokeratin in Plasma Cell Neoplasm

The Aberrant Expression of Cytokeratin in Plasma Cell Neoplasm

Inhibition of lung cancer in diethylnitrosamine-induced mice by Rhizoma paridis saponins.

Lung cancer is the foremost cause of cancer mortality and a growing economic burden worldwide. Rhizoma paridis saponins (RPS) have been reported to exhibit potential anti-tumor effects on many kinds of tumor models. The present study was designed to investigate the mechanism-based chemopreventive nature of RPS against DEN-induced lung carcinogenesis in Kunming mice. As a result, the treatment with RPS reduced the severity of pulmonary histopathology. The mechanism of its antitumor effect involved in (a) reducing oxidative stress injury through up-regulating activities of CAT and SOD; (b) down-regulating the levels of inflammatory factors, like TNF-α, IL6, COX-2, and PGE2; (c) activation of caspase-3 and up-regulating the pro-apoptotic protein Bax; (d) decreasing the expression of PCNA; (e) depressing the expression of cancer stem cells marker CD133; (f) suppressing aberrant expression of cytokeratin 8 and 18; and (g) inhibiting EGFR/ PI3 K/Akt, EGFR/Ras/Erk and NF-κB pathways. Taken together, RPS would be a potent agent inhibiting lung tumor in the future.

An unusual case of follicular dendritic cell sarcoma of the omentum with pleomorphous morphology and aberrant cytokeratin expression

Follicular dendritic cell sarcoma (FDCS) is an uncommon tumor arising from follicular dendritic cells (FDCs) in the lymph nodes or in extranodal sites. We herein report an unusual case of a 30-year-old man who presented with a FDCS of the omentum and massive liver involvement. The tumor proliferation was a diffuse growth composed by large and highly pleomorphic cells. On immunostaining, B and T cell markers; CD30, myeloid, and histiocytic markers; and S100 were negative whereas CD21, CD23, CD35, clusterin, and fascin were positive. Unexpectedly, large spectrum cytokeratin clone AE1/AE3 was focally and variably expressed. Despite the latter atypical finding and the misleading morphology, immunohistochemistry allowed for a conclusive diagnosis of FDCS.

Intravascular large B-cell lymphoma presenting as cauda equina syndrome and showing aberrant cytokeratin expression: a diagnostic challenge

Intravascular large B-cell lymphoma presenting as cauda equina syndrome and showing aberrant cytokeratin expression: a diagnostic challenge

Aberrant cytokeratin expression as a possible prognostic predictor in poorly differentiated colorectal carcinoma

The cytokeratin (CK)7(-) /CK20(+) immunoprofile is characteristic of colorectal carcinoma (CRC), although CK7(+) or CK20(-) phenotypes are occasionally encountered, particularly in histologically variant CRCs. We analyzed CK7/CK20 profiles in variant CRCs in association with clinicopathologic parameters and prognosis. CK expression in well- and moderately differentiated adenocarcinoma (WMDA) (n = 63), poorly differentiated adenocarcinoma (PDA) (n = 91), mucinous adenocarcinoma (MUA) (n = 81), signet-ring cell carcinoma (SRCC) (n = 15), undifferentiated carcinoma (UDC) (n = 12), and adenosquamous carcinoma (n = 2) was analyzed using immunohistochemistry. Cut-off scores were set at 1% for CK7 and 25% for CK20 using the receiver operating characteristic curve analysis of PDA. Association between CK20(-) and better prognosis in PDA was validated in the second cohort (n = 66). CK7/CK20 immunoprofiling revealed a predominant CK7(-) /CK20(+) profile in WMDA, MUA, and SRCC, while the majority of UDC was characterized by a CK7(-) /CK20(-) profile. The CK7/CK20 profile in PDA was variable. Contingency table analysis revealed that CK expression was not significantly associated with any clinicopathologic parameters in WMDA, PDA, and MUA. However, survival analysis demonstrated that CK20(-) was significantly associated with better prognosis in PDA. Although CK20(-) was significantly associated with mismatch repair deficiency in PDA, it was an independent prognostic factor in multivariate analysis. Finally, we confirmed that CK20 status, determined using a 25% cut-off score, was a significant prognostic parameter in the second PDA cohort. CK20 status may be used as a prognostic predictor of PDA.

MMP-9 and CXCL8/IL-8 are potential therapeutic targets in epidermolysis bullosa simplex.

Epidermolysis bullosa refers to a group of genodermatoses that affects the integrity of epithelial layers, phenotypically resulting in severe skin blistering. Dowling-Meara, the major subtype of epidermolysis bullosa simplex, is inherited in an autosomal dominant manner and can be caused by mutations in either the keratin-5 (K5) or the keratin-14 (K14) gene. Currently, no therapeutic approach is known, and the main objective of this study was to identify novel therapeutic targets. We used microarray analysis, semi-quantitative real-time PCR, western blot and ELISA to identify differentially regulated genes in two K14 mutant cell lines carrying the mutations K14 R125P and K14 R125H, respectively. We found kallikrein-related peptidases and matrix metalloproteinases to be upregulated. We also found elevated expression of chemokines, and we observed deregulation of the Cdc42 pathway as well as aberrant expression of cytokeratins and junction proteins. We further demonstrated, that expression of these genes is dependent on interleukin-1 β signaling. To evaluate these data in vivo we analysed the blister fluids of epidermolysis bullosa simplex patients vs. healthy controls and identified matrix metalloproteinase-9 and the chemokine CXCL8/IL-8 as potential therapeutic targets.

Kaposi sarcoma herpesvirus/human herpesvirus‐8–negative effusion‐based lymphoma: Report of 3 cases and review of the literature

Primary effusion lymphoma (PEL) is a rare subtype of large B-cell lymphoma that arises in body cavities without detectable tumor masses. PEL is universally associated with Kaposi sarcoma herpesvirus (KSHV)/human herpesvirus-8 (HHV8). Despite overlapping features, KSHV/HHV8-negative effusion-based lymphoma is a distinct entity from PEL. To date, 52 cases have been reported. The authors report 3 additional cases received in their laboratory from 2007 to 2012. Clinical data, cytomorphologic features, and immunophenotypic features of the 3 cases were described and compared with those reported in the literature. The cells in HHV8-negative effusion lymphoma commonly revealed large cell, immunoblastic morphology and B-cell immunophenotype. The 3 cases demonstrated cytomorphologic and immunophenotypic variability. Cytomorphologically, 1 case contained large, highly atypical cells with a moderate amount of cytoplasm, round nucleus, coarsely granular chromatin, and a single macronucleolus. The other 2 cases had medium to large atypical cells with high nuclear-to-cytoplasmic ratios, slightly irregular to cleaved nuclei, and multiple conspicuous nucleoli. One case had a null phenotype with aberrant cytokeratin expression. B-cell phenotype was established by clonal immunoglobulin heavy-chain rearrangement using polymerase chain reaction, whereas the other 2 cases demonstrated a B-cell phenotype by flow cytometry and immunohistochemical staining. All 3 cases were negative for both HHV8 and Epstein-Barr virus. HHV8-negative effusion lymphoma exhibits clinical, cytomorphologic, and immunophenotypic variability. Cases with a null-phenotype can be particularly challenging. When effusion lymphoma is suspected, ancillary tests are helpful. Moreover, HHV8 detection is critical in differentiating PEL and HHV8-negative effusion lymphoma, because they have overlapping features yet different prognoses.

Epithelial‐to‐mesenchymal transition: immunohistochemical investigation of related molecules in canine cutaneous epithelial tumours

Epithelial-to-mesenchymal transition (EMT) is a multistep process, important in tumour invasion and metastasis, characterized by loss of epithelial markers, redistribution of β-catenin and gain of mesenchymal markers. HYPOSTHESIS/OBJECTIVES: Our aim was to investigate the immunohistochemical aberrant expression of cytokeratin, vimentin, survivin and heat shock protein 72 (Hsp72) in canine cutaneous epithelial tumours, to understand the association of expression of these molecules with features of malignancy and their role in the EMT phenotype. Ten canine squamous cell carcinomas (SCCs; one with lymph node metastasis), 30 canine hair follicle tumours (six pilomatricomas, eight infundibular keratinizing acanthomas, six trichoepitheliomas and 10 trichoblastomas) and five normal skin samples were investigated by immunohistochemistry using specific anti-vimentin, -cytokeratin, -survivin and -Hsp72 antibodies. A semi-quantitative method was used to analyse the results, as follows: 0 to <5%; ≥ 5 to <10%; ≥ 10 to <25%; and ≥ 25% of positive cells. Immunofluorescence was performed to investigate survivin-vimentin and survivin-Hsp72 colocalization in selected SCCs. Results - In malignant hair follicle tumours and SCCs, a reduced intensity of cytokeratin and increased survivin and Hsp72 expression were observed. In SCCs, loss of cytokeratin expression and vimentin immunolabelling, suggestive of the EMT phenotype, were evident in <5% of neoplastic cells in the front of tumour invasion. In the same areas, strong nuclear survivin and cytoplasmic Hsp72 staining was evident, often colocalizing. Only a few neoplastic cells in the front of tumour invasion showed vimentin-survivin colocalization. A possible simultaneous involvement of survivin and Hsp72 in tumour invasion and the multistep process of EMT of cutaneous epithelial tumours of dogs is suggested.

Atypical fibroxanthoma with T-cytotoxic inflammatory infiltrate and aberrant expression of cytokeratin

Journal of Cutaneous PathologyVolume 38, Issue 11 p. 930-932 Atypical fibroxanthoma with T-cytotoxic inflammatory infiltrate and aberrant expression of cytokeratin Dimas Suárez-Vilela MD, Dimas Suárez-Vilela MD Department of Anatomía Patológica, Hospital Valle del Nalón, Polígono de Riaño, S/N, 33920 Riaño (Langreo) Asturias, SpainSearch for more papers by this authorFrancisco Miguel Izquierdo MD, Corresponding Author Francisco Miguel Izquierdo MD Department of Anatomía Patológica, Complejo Asistencial de León, Altos de la Nava s/n, 24008 León, Spaine-mail: [email protected]Search for more papers by this authorJuliana Escobar-Stein MD, Juliana Escobar-Stein MD Department of Anatomía Patológica, Hospital Valle del Nalón, Polígono de Riaño, S/N, 33920 Riaño (Langreo) Asturias, SpainSearch for more papers by this authorJose Ramón Méndez-Alvarez MD, Jose Ramón Méndez-Alvarez MD Department of Anatomía Patológica, Hospital Valle del Nalón, Polígono de Riaño, S/N, 33920 Riaño (Langreo) Asturias, SpainSearch for more papers by this author Dimas Suárez-Vilela MD, Dimas Suárez-Vilela MD Department of Anatomía Patológica, Hospital Valle del Nalón, Polígono de Riaño, S/N, 33920 Riaño (Langreo) Asturias, SpainSearch for more papers by this authorFrancisco Miguel Izquierdo MD, Corresponding Author Francisco Miguel Izquierdo MD Department of Anatomía Patológica, Complejo Asistencial de León, Altos de la Nava s/n, 24008 León, Spaine-mail: [email protected]Search for more papers by this authorJuliana Escobar-Stein MD, Juliana Escobar-Stein MD Department of Anatomía Patológica, Hospital Valle del Nalón, Polígono de Riaño, S/N, 33920 Riaño (Langreo) Asturias, SpainSearch for more papers by this authorJose Ramón Méndez-Alvarez MD, Jose Ramón Méndez-Alvarez MD Department of Anatomía Patológica, Hospital Valle del Nalón, Polígono de Riaño, S/N, 33920 Riaño (Langreo) Asturias, SpainSearch for more papers by this author First published: 23 August 2011 https://doi.org/10.1111/j.1600-0560.2011.01774.xCitations: 5Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume38, Issue11November 2011Pages 930-932 RelatedInformation

Prognostic value of tissue polypeptide antigen in oral squamous cell carcinoma

Human oral cancer has a high risk of locoregional relapse which is difficult to diagnose early due to lack of prognostic markers. We and others have shown aberrant expression of cytokeratin (CK) 8 and 18 in human oral cancer. Aberrant supra-basal expression of CK19 has been shown earlier. In this study, our aim was to develop a non-invasive test for prognostication of human oral cancer. Immunoradiometric assay (IRMA) was used to measure the circulating levels of TPA in the sera of 80 oral cancer patients before surgery and seven days after surgery. Elevated serum TPA levels were noted in the post surgery samples of 28 out of the 50 patients for whom clinical follow-up was available, as compared to their pre-surgery samples. TPA levels in the pre-surgery serum samples of patients were significantly higher than levels in the sera of healthy controls (p=0.001). Elevated levels in patients correlated significantly with stage (p=0.02), development of recurrence (p<0.006), and impacted survival (p<0.033). IHC analysis showed statistically significant correlation between expression of CK8, 18 and 19 in surrounding uninvolved tissues of the tumour and post surgery elevated serum TPA levels (p<0.001). This suggests the possibility that CK fragments are released from the surrounding uninvolved tissues into the sera of patients after surgical removal of the tumour. Thus, our study indicates that TPA can be a useful tumour marker for the prediction of recurrence and poor prognosis in human oral cancer.

Aberrant expression of cytokeratin 7 in perivenular hepatocytes correlates with a cholestatic chemistry profile in patients with heart failure

A cholestatic liver chemistry profile (elevations in alkaline phosphatase and bilirubin) is commonly encountered in patients with hepatic venous outflow obstruction due to right heart failure. Liver biopsies from these patients demonstrate varying degrees of sinusoidal dilatation, red blood cell extravasation and sinusoidal congestion. Recently, a bile ductular reaction mimicking biliary tract disease has been identified in ∼50% of patients with venous outflow obstruction possibly explaining the cholestatic profile encountered in these patients. In this study, we evaluated the liver biopsies from 22 patients with heart failure. Marked sinusoidal dilatation involving zones 2 and 3 was observed in 15 patients. Similar to a previous study, 7 of 22 patients had histologic evidence of a mild ductular reaction. Cytokeratin 7 immunohistochemistry revealed a mild ductular reaction in an additional two cases. Strikingly, CK7 was aberrantly expressed in perivenular hepatocytes in 20 of 22 cases. Perivenular CK7 immunoreactivity was focal in most cases; however, in five cases it was quite extensive and extended into zone 2. There was no significant association between marked sinusoidal dilatation and extensive perivenular CK7 positivity. Extensive perivenular CK7 positivity was significantly associated with both elevated bilirubin, as well as the presence of fibrosis. However, a ductular reaction was not associated with a cholestatic liver chemistry profile.

Aberrant cytokeratin expression during arsenic-induced acquired malignant phenotype in human HaCaT keratinocytes consistent with epidermal carcinogenesis

Inorganic arsenic is a known human skin carcinogen. Chronic arsenic exposure results in various human skin lesions, including hyperkeratosis and squamous cell carcinoma (SCC), both characterized by distorted cytokeratin (CK) production. Prior work shows the human skin keratinocyte HaCaT cell line, when exposed chronically for >25 weeks to a low level of inorganic arsenite (100 nM) results in cells able to produce aggressive SCC upon inoculation into nude mice. In the present study, CK expression analysis was performed in arsenic-exposed HaCaT cells during the progressive acquisition of this malignant phenotype (0–20 weeks) to further validate this model as relevant to epidermal carcinogenesis induced by arsenic in humans. Indeed, we observed clear evidence of acquired cancer phenotype by 20 weeks of arsenite exposure including the formation of giant cells, a >4-fold increase in colony formation in soft agar and a ∼2.5-fold increase in matrix metalloproteinase-9 secretion, an enzyme often secreted by cancer cells to help invade through the local extra-cellular matrix. During this acquired malignant phenotype, various CK genes showed markedly altered expression at the transcript and protein levels in a time-dependent manner. For example, CK1, a marker of hyperkeratosis, increased up to 34-fold during arsenic-induced transformation, while CK13, a marker for dermal cancer progression, increased up to 45-fold. The stem cell marker, CK15, increased up to 7-fold, particularly during the later stages of arsenic exposure, indicating a potential emergence of cancer stem-like cells with arsenic-induced acquired malignant phenotype. The expression of involucrin and loricrin, markers for keratinocyte differentiation, increased up to 9-fold. Thus, during arsenic-induced acquired cancer phenotype in human keratinocytes, dramatic and dynamic alterations in CK expression occur which are consistent with the process of epidermal carcinogenesis helping validate this as an appropriate model for the study of arsenic-induced skin cancer.

Molecular and phenotypic analysis of poorly differentiated sinonasal neoplasms: an integrated approach for early diagnosis and classification

Primary poorly differentiated (small round and non-small) sinonasal neoplasms comprise histogenetically and biologically diverse entities with overlapping morphologic features. Because of the limited initial biopsy tissue materials, differential diagnostic difficulties may arise and complicate timely management of some cases. We used immunohistochemical and molecular marker analyses in a large cohort of these tumors to optimize their early diagnosis and classification. Fifty-two tumors of the skull base and sinonasal regions and, for comparison, 19 poorly differentiated neoplasms of other head and neck sites were analyzed by a panel of immunohistochemical markers including those of epithelial, mesenchymal, melanocytic, and neuroectodermal origin using tissue microarray. Reverse transcriptase-polymerase chain reaction analysis of messenger RNA for EWS-FLI1 and PAX-FKHR fusion transcripts and the human achaete-scute homolog-1 gene was performed on 24 of the 52 sinonasal tumors and the 19 tumors of other sites for comparison. The immunohistochemical results substantiated the phenotypic assessment and the initial diagnosis in 49 of the 52 tumors. In 4 instances the integrated markers and phenotypic analyses led to reclassification of 3 tumors and confirmed the histogenesis of a mesenchymal tumor with aberrant cytokeratin expression. Molecular analysis of the EWS-FLI1 fusion gene transcript revealed 4 (9.3%) of the 43 tumors to be positive; all were Ewing sarcomas. The human achaete-scute homolog-1 gene transcript was identified in 10 (23.8%) of 42 tumors: 3 of 6 neuroblastomas, all 4 neuroendocrine carcinomas, and 1 each in sinonasal undifferentiated carcinoma, rhabdomyosarcoma, and melanoma. The PAX-FKHR fusion transcript was not detected in any tumors. We conclude that (1) an integrated morphologic and biomarker algorithm may better optimize the early diagnosis of poorly differentiated sinonasal and skull-base tumors; (2) molecular analysis may assist in future biological stratification of certain classes of these tumors; and (3) the human achaete-scute homolog-1 gene transcript is a nonspecific marker for the diagnosis of neuroblastoma.