Comparison of Cytogenetic and Static Cytometry Procedures in the Evaluation of Potentially Malignant Oral Lesions

Gross genomic damage or specific chromosomal alterations have been revealed by different laboratory procedures in potentially malignant oral lesions, but two or more procedures have never been applied at the same time to the same cell population. In the present study we considered cell suspensions obtained from 34 oral lesions at risk of malignancy to see whether they might harbour genetic alterations and whether a correlation exists between the results obtained by two different methods of assessing DNA aberrations. Each suspension underwent DNA-content assessment by static cytometry, and cytogenetic G-banding analysis of short-term primary cultures. DNA content was determined in a minimum of 1000 cells on a Fairfield ploidy analyser and results expressed as percent of aneuploid cells in the S-phase; cytogenetic analysis was carried out according to standard procedures on in situ G-banding metaphases, and results expressed as percent of metaphases with chromosomal alterations. The results showed that the percentage of metaphases with chromosomal alterations was significantly correlated with the percentage of aneuploid cells in the S-phase. In conclusion, genetic alterations can be revealed in the same oral specimen either by procedures studying DNA content in fixed cells or by procedures investigating chromosomal alterations in cultured and proliferating cells

The potential role of the extracellular matrix in the activity of trabectedin in UPS and L-sarcoma: evidences from a patient‐derived primary culture case series in tridimensional and zebrafish models

Background: Soft tissue sarcomas (STS) are a rare group of solid neoplasm including among others liposarcoma, leiomyosarcoma (L-sarcoma) and undifferentiated pleomorphic sarcoma (UPS) entities. The current first-line treatment is represented by anthracycline based- regimens, second-line may include trabectedin. Currently the activity of trabectedin and its mechanism of action is not completely elucidated. Methods: Taking the advantages of our 3D patient-derived primary culture translational model we performed genomic-, chemobiogram, proteomic- and in vivo analysis in a UPS culture (S1). Furthermore pharmacological profiling of a UPS and L-sarcoma patient-derived case series and in silico analysis were carried out. Results: Trabectedin exhibited an increased activity in 3D respect to 2D cultures suggesting an extracellular matrix (ECM) and timp1 involvement in its mechanism of action. Moreover 3D S1 xenotranspanted zebrafish model showed an increased sensitivity to trabectedin. Finally the results were further validated in a UPS and L-sarcoma case series. Conclusions: Taken together these results confirmed the activity of trabectedin in these STS histotypes. Moreover the data underline the ECM involvement in the cytotoxic effect mediated by trabectedin and could open the door for researches aimed to focus on the patient setting that could benefit from this agent

CITOGENETICA DEI CARCINOMI POLMONARI NON A PICCOLE CELLULE IN FUMATORI

Introduzione: Le alterazioni genetiche piu\u2019 frequenti nel carcinoma polmonare non a piccole cellule (CNPC) sono rappresentate da perdite del 3p, sede del gene FHIT, e varie alterazioni numeriche a carico dei cromosomi 9, 17, 21 e 22. tuttavia la maggior parte delle casistiche presenti in letteratura sono state studiate mediante FISH o ricerca della perdita di eterozigosi. Pochi sono i dati attuali sulla determinazione del cariotipo completo, con citogenetica classica in metafase su ampie casistiche. Metodi: Il cariotipo di 38 CNPC del polmone \ue8 stato studiato attraverso la tecnica delle colture a breve termine e l\u2019analisi su piastre metafasiche. Risultati: La casistica \ue8 rappresentata da 19 carcinomi squamocellulari (SC), 18 adenocarcinomi (AC) e 1 carcinoma a grandi cellule (CGC). Nei CS le alterazioni piu\u2019 frequenti osservate erano alterazioni numeriche, in particolare, a carico del cromosoma 21 ( in 6 casi) e l\u2019acquisizione di un cromosoma 3 (in 9 casi: clonale in 2 e come singola metafase in 7). L\u2019acquisizione del cromosoma 3 (trisomia 3) \ue8 stata riscontrata, sia associata alla perdita del 21, sia come unica alterazione. Conclusioni: Mentre la monosomia 21 \ue8 un fenomeno ben conosciuto nelle cellule tumorali squamose, piu\u2019 sorprendente appare nella nostra casistica il riscontro di trisomia 3. Infatti, nel braccio corto del cromosoma 3 risiede il gene FHIT la cui perdita \ue8 descritta come evento precoce nella carcinogenesi polmonare. Al fine di confermare i risultati ottenuti, i casi sono stati testati mediante immunoistochimica con anticorpo anti-FHIT e casi selezionati sono stati sottoposti anche a FISH con sonda centromerica per il cromosoma 3

IGFBP2 as an Immunohistochemical Marker for Prostatic Adenocarcinoma

AIM: The aim of the study was to evaluate the immunohistochemical expression of insulin-like growth factor binding protein 2 (IGFBP2) in normal epithelium, high-grade prostatic intraepithelial neoplasia (HG-PIN), and prostatic adenocarcinoma (PAc), in patients hormonally untreated and in those having undergone complete androgen ablation. MATERIALS AND METHODS: IGFBP2 expression was evaluated in PAc, HG-PIN, and normal-appearing epithelium in 40 radical prostatectomies from hormonally untreated patients and 10 radical prostatectomies from patients under complete androgen ablation before surgery. The study also included the initial biopsies of such patients, and an additional 10 simple prostatectomies from patients with bladder outlet obstruction. Statistics included receiver-operator characteristic curves, the Wilcoxon test, and the Spearman test. Results were compared with \u3b1-methylacyl-CoA racemase. RESULTS: The principal findings were: (1) IGFBP2 was not expressed in normal ducts and acini of the transition and peripheral zones; (2) IGFBP2 was expressed in the cytoplasm of untreated PAc and, to a lesser extent, in HG-PIN; (3) it was also expressed in PAc and HG-PIN after complete androgen ablation, but to a lesser extent than in the untreated neoplasms; (4) \u3b1-methylacyl-CoA racemase was expressed both in PAc and HG-PIN, the level being similar in both lesions and lower in the specimens from the patients having undergone androgen ablation. CONCLUSIONS: Results obtained show that IGFBP2 is expressed in invasive PAc, whereas its expression in HG-PIN is low. These findings can be helpful in the correct diagnosis of PAc both in biopsies and in surgical specimens, mainly in untreated patients

Immunohistochemical Expression of the Human Sodium/Iodide Symporter Distinguishes Malignant From Benign Gastric Lesions

AIM: Sodium/iodide symporter (NIS) is a transmembrane protein that mediates the transport of I(-). The aim was to evaluate the immunohistochemical expression of the human homolog of NIS (hNIS) in a wide spectrum of gastric lesions. MATERIALS AND METHODS: Seventy-seven samples were stained immunohistochemically with a monoclonal antibody for hNIS, including 14 with normal gastric mucosa, 14 with chronic atrophic gastritis with foveolar hyperplasia, 15 with chronic atrophic gastritis with intestinal metaplasia, 6 with chronic atrophic gastritis with atypical regenerative hyperplasia, 8 with chronic atrophic gastritis with dysplasia, 15 with invasive adenocarcinoma, 3 with well-differentiated neuroendocrine tumor, and 2 with gastrointestinal stromal tumors (GISTs). RESULTS: hNIS stained the basolateral cytoplasmic portion of foveolae in normal mucosa, in 13 cases of chronic atrophic gastritis with foveolar hyperplasia, and in only 1 case of regenerative atypical hyperplasia. hNIS was consistently absent in intestinal metaplasia, in dysplastic glands, and in the cells constituting invasive carcinoma, well-differentiated neuroendocrine tumors, and GIST. CONCLUSION: It seems that lack of hNIS can be useful in distinguishing foveolar hyperplasia from dysplastic glands

LATE SKIP LYMPH NODE METASTASIS OF ORAL SQUAMOUS CELL CARCINOMA OR METASTASIS OF UNKNOWN SECOND PRIMARY TUMOR?ANSWER BY MITOCHONDRIAL DNA ANALYSIS

BACKGROUND: Histological and clinical criteria are generally used to differentiate second primary tumors (SPTs) from local recurrences. The purpose of the present study was to apply mitochondrial DNA (mtDNA) D-loop analysis to differentiate SPTs from local recurrences and to validate the clinical classification. METHODS: The study population consisted of 20 consecutive patients presenting multiple oral neoplastic lesions for a total of 25 paired lesions. The mtDNA D-loop analysis was performed by direct sequencing and phylogenetic clusterization. RESULTS: Agreement between mtDNA analysis and clinical classification was found in 19 cases. Discrepancies arose in 6 cases in which the clinical criteria based only on the spatial or temporal distance of the second lesion from the index tumor had led to a diagnosis of SPT (2 cases) or local recurrence (4 cases). CONCLUSION: The present data highlight the value of mtDNA analysis in establishing the clonal relationship between the index tumor and the second neoplastic lesion. \ua9 2013 Wiley Periodicals, Inc. Head Neck 36: 94-100, 2014