<span title="">2014</span> <i title="Springer Nature"> <a target="_blank" rel="noopener" href="https://fatcat.wiki/container/l7jwciowjvhanbf3cedbhw5g2a" style="color: black;">Modern Pathology</a> </i> &nbsp;
332A ANNUAL MEETING ABSTRACTS benign categories was 100%. 125/125 (100%) of cases had HR and 80/125 (64%) for LR HPV ISH testing done. Malignant diagnoses (n=74) included 55 keratinizing SCC (KSCC);14 non-keratinizing SCC (NKSCC); 3 papillary SCC (PSCC);1 undifferentiated carcinoma;1 SCC in situ (SCCIS). Benign diagnoses (n=51) included 37 squamous papilloma (SP);6 squamous hyperplasia (SH);3 normal epithelium (NE);2 laryngeal nodule (LN);1 sinonasal papilloma (SNP); and 1 each of condyloma and
more &raquo; ... seborrheic keratosis (C/SebK) (last two are only samples outside of HN (inguinal)). 74/74 (100%) of malignant cases had HR HPV ISH done (26/55 of KSCC, 9/14 NKSCC, 3/3 of PSCC, 0/1 of SCCIS, and 0/1 of undifferentiated carcinoma positive), 34/74 (46%) malignant cases had LR HPV ISH done (0/21 of KSCC, 2/11 of NKSCC, 0/1 of PSCC, 0/1 of undifferentiated carcinoma positive). 51/51 (100%) of benign cases had HR HPV ISH done (2/37 of SP, 1/6 of SH, 0/3 of NE, 0/1 of SNP, 0/2 of LN, 0/2 of C/SebK positive). 46/51 (90%) of benign diagnoses had LR HPV ISH ordered (16/34 of SP, 0/4 of SH, 0/2 of LN, 0/1 of SNP, 0/3 of NE, 1/2 of C/SebK positive). Conclusions: Unnecessary HPV ISH testing in HN lesions is common. 46% HN carcinomas had LR HPV ISH and 100% of non-neoplastic and benign lesions had HR HPV ISH ordered. Our laboratory will discontinue offering HPV ISH Panel (HR+LR) in an attempt to reduce redundant testing.Training of clinicians and pathologists is essential in avoiding unnecessary costly testing. 1367 P16 Ink4A : No Correlation with Transcriptionally Active HPV16/18 or Outcomes in Oral Cavity Carcinoma J Xu, T Isayeva, M Brandwein-Gensler. University of Alabama at Birmingham, Birmingham, AL. Background: Transcriptionally active high-risk Human Papillomavirus (HR-HPV) is important in promoting oropharyngeal carcinomas (OPC) through binding of E6 and E7 viral oncoproteins with p53 and Rb tumor suppressor proteins, respectively, leading to inactivation. Loss of negative feedback secondary to Rb inactivation results in p16 Ink4A deregulation; p16 Ink4A overexpression is considered indicative of transcriptionally active and biologically relevant HPV infection. Recent studies suggest that p16 Ink4A overexpression alone, independent of HPV status, confers improved prognosis of OPC. Here we examine: (1) p16 Ink4A as a surrogate HPV biomarker in oral cavity carcinomas (OCC) and (2) whether p16 Ink4A and/or transcriptionally active HR-HPV can impact patient outcome for OCC. Design: 151 patients with OCC were identifi ed; patients with microinvasive carcinomas ( 4 mm) were excluded. RNA was extracted from archival specimens and reverse transcription was performed; residual DNA was removed by DNase digestion. Nested quantitative real-time PCR was performed with primers specifi c to HPV16 and HPV18 (E6/E7), respectively. Immunohistochemistry for p16 Ink4A expression was examined on whole tissue slides and OCC were classifi ed as positive if strong, diffuse nuclear and cytoplasmic p16 Ink4A expression was seen ( +2 intensity,≥ 75% distribution). Data on demographics and outcome were collected. Fisher's exact test was used to analyze HPV status and demographics; Kaplan Meier curves were used to analyze HPV status and outcome. Results: HPV16+ OCC was seen in 36/151 (24%) cases, HPV18+ in 12/151 (8%) cases, and no double HPV16/18 infections were found. p16 Ink4A overexpression was demonstrated in 13/84 (15%) OCC, and did not correlate with either HPV16 or HPV18 status. Neither p16 Ink4A expression nor HPV status were signifi cantly associated with overall survival, disease specifi c survival, or disease-free survival. Conclusions: Our data suggests that, in contrast to OPC, p16 Ink4A overexpression is not a surrogate marker for transcriptionally active HPV in OCC. The lack of impact of p16 Ink4A overexpression with OCC outcome may be due to the small numbers of p16 Ink4A overexpressors found (15% or 13 patients). Background: V-ATPase is involved in the acidifi cation of the microenvironment around/ in solid tumors, such as oral squamous cell carcinoma(OSCC). V-ATPase is thought to induce tumor invasion and multi-drug resistance in several malignant tumors. However, there is little information regarding the effects of V-ATPase inhibitors on OSCCs. Design: We attempted to assess the effect of V-ATPase inhibitor, concanamycin A1 (CMA), on cell proliferation and apoptosis of OSCC cells (MISK81-5, SAS, HSC-4 and SQUU-B) in vitro. The effects of CMA on the cell viability and apoptosis were investigated by MTS assay and TUNEL staining, respectively. The mRNA and protein expression levels of apoptosis-related molecules after CMA treatment were analyzed by qRT-PCR and Western blotting, respectively. Results: CMA treatment for 48 hr signifi cantly suppressed the cell growth at low concentrations, and induced apoptosis in MISK81-5, SAS, and HSC-4 cells, but SQUU-B cells were highly resistant to CMA. Compared the expression of the proand anti-apoptotic factors in the SQUU-B cells with that in CMA-sensitive OSCC cells after treatment with CMA, whereas CMA activated p38, one of MAPK, in the CMA-sensitive OSCC cells, phosphorylation of p38 was not observed in SQUU-B cells. Moreover, CMA treatment induced comparative increase in Bcl-2 expression in the SQUU-B cells compared with that in the CMA-sensitive cells. However, when the SQUU-B cells were treated with CMA and a histone deacetylase inhibitor, suberoylanilide hydroxamic acid (SAHA), the SQUU-B cells became more susceptible to the CMA-induced apoptosis. SAHA treatment led to a signifi cantly decreased Bcl-2 expression levels in comparison with that observed in the SQUU-B cells treated with CMA alone. Down-regulation of Bcl-2 partially induced decrease of the cell viability in the SQUU-B cells treated with CMA. Conclusions: Our results indicate that CMA could have an anti-tumor effect on OSCCs and that the combination of CMA with SAHA may exhibit synergistic anti-cancer activity in certain CMA resistance cells. Background: Alcohol use is one of the primary risk factors for head and neck squamous cell carcinoma (HNSCC), yet the mechanism by which alcohol induces oropharyngeal cancer remains unclear. Although ethanol itself is not carcinogenic, acetaldehyde, or ethanal, the fi rst metabolite of alcohol via the enzyme alcohol dehydrogenase, has been established as a suspected carcinogen. Long non-coding RNAs (lncRNAs) constitute a family of RNA longer than 200 nucleotides that do not code for proteins, but instead infl uence transcription factors associated with regulation of oncogenes, tumor suppressor proteins, self-renewal, and differentiation. This study sought to determine those key lncRNAs whose dysregulation by ethanol result in the acquisition of cancer stem cell properties in normal oral epithelial cells. Design: Two normal oral keratinocyte cell lines (OKF4 and OKF6) were treated with various doses of 100% ethanol for 28 days to represent long-term alcohol use. The same cell lines were also treated with acetaldehyde for 72 hours to better investigate the in vivo mechanism of alcohol-induced HNSCC. Relative lncRNA and mRNA gene expression in ethanol treated cells were compared with that of parental cells through qPCR arrays. Results: Comparison of relative lncRNA expression in ethanol treated cells with that of parental cells revealed that six lncRNAs (SAF, Zeb2Nat, Air, H19, IPW, and ncr-UPAR) are consistently upregulated. The expression of SAF and Zeb2Nat, two lncRNAs implicated in oncogenesis, exhibited the highest fold change, thus suggesting that SAF and Zeb2Nat are instrumental in the pathogenesis of alcohol-induced oropharyngeal cancers. The stem cell genes Nanog and Oct-4, and the epithelial-to-mesenchymal transition (EMT) gene Vimentin, were also upregulated through ethanol treatment, thereby suggesting that ethanol could promote expression of the EMT phenotype and stemness in vivo. Conclusions: Ethanol exposure in normal oral epithelia dysregulates key lncRNAs previously implicated in cancer, and induces stem cell and EMT genes. Further investigation of the pathways of these lncRNAs may lead to prognostic indicators and therapeutic targets in treatment of alcohol-induced oropharyngeal cancers. Background: We have previously demonstrated that reduced CXCR4 expression is associated with a lymphoplasmacytoid immunophenotype and extramedullary disease in a mouse model of plasma cell myeloma (PCM). We and others have also shown that reduced CXCR4 is associated with poor survival in PCM patients, especially those treated with bortezomib. In this study we sought to optimize CXCR4 immunohistochemical staining of plasma cells in formalin fi xed trephine core biopsies. In addition, we examined the association of CXCR4 expression with the cytogenetic and morphologic subtypes of PCM. We hypothesized that cases with a t(11;14) were more likely to have decreased/absent CXCR4 expression. Design: We queried our cytogenetics database and identifi ed 40 cases of PCM (including 24 diagnostic specimens) that harbored the most common recurrent cytogenetic abnormalities. We then reviewed bone marrow biopsies/smears and clinical data corresponding to the same accession dates. In addition, immunohistochemical (IHC) studies were performed on the core biopsies including CD20 and CXCR4. Stain intensity was scored from 0 (negative) to 3 (strongly positive). Statistical analysis was performed using Fisher's exact tests. Results: 35% of PCM cases (14/40) were positive for surface expression of CXCR4, while nearly all control plasma cells were positive. Similar to previous studies, CD20 positivity was associated with the presence of t(11;14) (P < 0.001, Fisher's), but was not associated with strong or absent CXCR4 expression. Among the cases that bore a t(4;14) or t(14;16), all lacked CXCR4 expression (P = 0.035, Fisher's). In addition, among all cytogenetic abnormalities, cases without hyperdiploidy were more likely to lack CXCR4 (P =.041, Fisher's). Conclusions: Our previous studies have demonstrated that bortezomib resistant PCM cells are more likely to have an immunophenotype intermediate between lymphocytes and plasma cells, including decreased CXCR4. We also have shown that reduced CXCR4 expression confers a worse prognosis in PCM patients. Our data confi rm that PCM cells harboring a t(11;14) are more likely to express CD20 but do not appear to have ANNUAL MEETING ABSTRACTS 333A an association with CXCR4 expression. However, cells that have a non-hyperdiploid karyotype, including t(4;14) or t(14;16) are much more likely to lack CXCR4. These data may suggest that decreased CXCR4 can occur independent of a lymphoplasmacytoid immunophenotype. We propose that CXCR4 in PCM can be characterized by IHC, but further study will be necessary to determine whether CXCR4 can be an independent marker of prognostic information. The Utility of Morphology, Immunohistochemistry, Flow Cytometry and FISH Analysis in Assessment of Plasma Cell Neoplasm in the Bone Marrow OE Ajise, M Roshal, GN Sukhram, J Rueda, KM Smith, P Maslak, A Dogan. Memorial Sloan-Kettering Cancer Center, New York, NY. Background: Bone marrow (BM) evaluation of plasma cell (PC) disorders requires enumeration of PC and demonstration of clonality. Enumeration of the PCs can be performed in the BM aspirate, by immunohistochemistry (IHC) for CD138 in the BM biopsy, or by fl ow cytometry (FC). Clonality can be demonstrated by establishing light chain (LC) restriction of the PC in BM biopsy by IHC, LC restriction and/or abnormal phenotype by FC or cytogenetic abnormalities. A direct comparison of the utility of multiple testing modalities in accomplishing these goals has not been previously reported from a US-based study, we sought to compare these modalities for the evaluation of PC neoplasms. Design: 100 consecutive BM samples submitted for evaluation of PC neoplasms were studied through HE stained biopsy cores, IHC for CD138 and LC, Wright-Giemsa stained aspirates and highly sensitive FC. Multiple myeloma (MM)-specifi c fl uorescence in situ hybridization (FISH) was performed on PC-enriched BM cells. All 100 samples had morphology with corresponding FC analysis, 95 had an evaluable aspirate smear and 86 had myeloma FISH. Morphologic evaluation was judged positive when greater than 5% PC were present in the BM aspirate or biopsy and clonality was established by IHC. FC positivity required demonstration of at least 50 LC-restricted PCs showing abnormal immunophenotype. FISH was judged positive when MM specifi c chromosomal abnormalities were detected. Results: 93 patients had an established diagnosis of PC neoplasm, while 7 cases were new submissions. Of the 100 samples, 81 demonstrated clonal PC proliferation. CD138 stain yielded the highest estimates of the plasma cell proportion compared to aspirate count or fl ow cytometry (p<0.01) with mean values of 30% (range: 3-100), 19 (1-78)  2.1 (0.01-33.2) respectively. The biopsy proportion estimate correlated well with the aspirate count (r 2 =0.6), but poorly with FC evaluation (r 2 = 0.1). FC demonstrated the highest sensitivity in assessment of clonality 96% (78/81), while morphology+IHC was positive in 84% (68/81), followed by MM FISH/Cytogenetics 79% (59/75). Two cases were positive on morphology while negative by FC due to clotted FC samples. Conclusions: Our results show that IHC for CD138 is the most sensitive method for assessment of PC numbers in the BM specimens involved by a PC neoplasm. In contrast FC immunophenotyping is the most sensitive method to establish the clonal nature of the BM PC. Background: MicroRNAs (miRNA) signatures are shown to exert an important role in patho-biology and prognosis of acute myeloid Leukemia (AML). miR-223, is expressed at low levels in hematopoietic stem cells, but its expression increases dramatically during granulocytic differentiation. High levels of miR-223 lead to granulocytic differentiation in APML cells lines exposed to ATRA treatment. miR-223 exhibits its antiproliferative effects by targeting the transcription factors like MEF2C, that is up-regulated in leukemic monocyte. In this study, we examined the expression of miR-223 in a large cohort of AML patients (pts). and correlated it with cytogenetic data, various morphological sub-types (FAB) and over-all survival (OS). Design: Diagnosis /classifi cation (FAB, WHO 2008) was based on morphology, fl owcytometry and conventional cytogenetics / FISH studies. Diagnostic bone marrow biopsy tissue (FFPE) was utilized to extract microRNA (Qiagen). miScript SYBR Green PCR Kit (Qiagen) was used to amplify the target (miR-223). Normal BM samples and peripheral blood mononuclear cells were used as normal controls. ddCt algorithm was used to analyze relative changes in gene expression. SPSS (version 20.0) was used for statistical analysis. Results: 117 pts. (18-89 yrs, mean 55 yrs, median 59 yrs, 73 men and 44 women, M:F 1.6:1) were included. Morphologically pts were classifi ed as, M0/M1 (23/117;20%); M2 (19/117,16%); M3 (11/117,9%); M4/M5 (31/117,26%); M6/M7 (8/117,7%) and others (25/117, 21%). Diploid karyotype was seen in 38/117(32%) while aneuploidy in 79/117(68%). miR-223 expression was higher (>1.2 fold of normal ) in only 28/117 (24%). miR-223 expression was higher in AML with diploid karyotype, compared to AML with aneuploidy (37% vs. 16%) (p<0.0194). Higher expression of miR-223 was noted most frequently in AML with differentiation (FAB-M2) (12/20; 60%) compared to M0/M1 (13% p<0.0069); M3 (0% ; p <0.0014); M4/M5 (17% p <0.0012); AML-MDS (21%; p<0.0312). miR-223 showed no correlation with over-all survival in normal vs. abnormal cytogenetic sub-groups or between any FAB subtypes. Conclusions: In AML, myelopoiesis regulator miR-223 expression is up-regulated in only a small subset of pts. Higher miR-223 expression is associated with normal cytogenetics and AML associated with differentiation (FAB-M2). miR-223 high expression has no impact on over-all survival in adult AML.
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