Integrated Analysis of miRNA and mRNA Endorses a Twenty miRNAs Signature for Colorectal Carcinoma.

Colorectal cancer (CRC) ranks as the most frequent carcinoma worldwide. CRC patients show strong prognostic differences and responses to treatment, and 20% have incurable metastatic disease at diagnosis. We considered it essential to investigate mechanisms that control cellular regulatory networks, such as the miRNA-mRNA interaction, known to be involved in cancer pathogenesis. We conducted a human miRNome analysis by TaqMan low density array, comparing CRC to normal colon tissue (NCT, and experimentally identified gene targets of miRNAs deregulated, by anti-correlation analysis, with the CRC whole-transcriptome profile obtained from RNASeq experiments. We identified an integrated signature of 20 deregulated miRNAs in CRC. Enrichment analyses of the gene targets controlled by these miRNAs brought to light 25 genes, members of pathways known to lead to cell growth and death (CCND1, NKD1, FZD3, MAD2L1, etc.), such as cell metabolism (ACSL6, PRPS1-2). A screening of prognosis-mediated miRNAs underlined that the overexpression of miR-224 promotes CRC metastasis, and is associated with high stage and poor survival. These findings suggest that the biology and progression of CRC depend on deregulation of multiple miRNAs that cause a complex dysfunction of cellular molecular networks. Our results have further established miRNA-mRNA interactions and defined multiple pathways involved in CRC pathogenesis.

Clinical and pathological factors influencing survival in a large cohort of triple-negative breast cancer patients.

BACKGROUND: To provide further information on the clinical and pathological prognostic factors in triple-negative breast cancer (TNBC), for which limited and inconsistent data are available. METHODS: Pathological characteristics and clinical records of 841 TNBCs diagnosed between 1994 and 2015 in four major oncologic centers from Sardinia, Italy, were reviewed. Multivariate hazard ratios (HRs) for mortality and recurrence according to various clinicopathological factors were estimated using Cox proportional hazards models. RESULTS: After a mean follow-up of 4.3 years, 275 (33.3%) TNBC patients had a progression of the disease and 170 (20.2%) died. After allowance for study center, age at diagnosis, and various clinicopathological factors, all components of the TNM staging system were identified as significant independent prognostic factors for TNBC mortality. The HRs were 3.13, 9.65, and 29.0, for stage II, III and IV, respectively, vs stage I. Necrosis and Ki-67 > 16% were also associated with increased mortality (HR: 1.61 and 1.99, respectively). Patients with tumor histotypes other than ductal invasive/lobular carcinomas had a more favorable prognosis (HR: 0.40 vs ductal invasive carcinoma). No significant associations with mortality were found for histologic grade, tumor infiltrating lymphocytes, and lymphovascular invasion. Among lymph node positive TNBCs, lymph node ratio appeared to be a stronger predictor of mortality than pathological lymph nodes stage (HR: 0.80 for pN3 vs pN1, and 3.05 for >0.65 vs <0.21 lymph node ratio), respectively. Consistent results were observed for cancer recurrence, except for Ki-67 and necrosis that were not found to be significant predictors for recurrence. CONCLUSIONS: This uniquely large study of TNBC patients provides further evidence that, besides tumor stage at diagnosis, lymph node ratio among lymph node positive tumors is an additional relevant predictor of survival and tumor recurrence, while Ki-67 seems to be predictive of mortality, but not of recurrence.

Contrast-enhanced ultrasonographic evaluation of inflammatory activity in Crohn's disease.

BACKGROUND & AIMS: We sought to test the diagnostic accuracy of ultrasound (US), color Doppler US (CD-US), and contrast-enhanced US (CE-US) in the evaluation of inflammatory activity in patients with Crohn's disease (CD), and to correlate the findings of these sonographic studies with inflammatory activity, as scored by the CD activity index (CDAI). METHODS: Patients with CD were enrolled in the study. Radiologists performing the scans were blinded to clinical status. Baseline US, CD-US, and CE-US examinations were conducted with high-frequency probes (8-14 and 5-7 MHz) before and after injection of sulfur hexafluoride-filled microbubbles. The diagnostic accuracy of baseline US, CD-US, and CE-US were calculated by using the endoscopic and histologic findings as reference standards and correlated with the CDAIs by using the Pearson linear correlation coefficient. RESULTS: Forty-seven patients (20 men; 27 women; mean age +/- SD, 38 +/- 14 years) with a CDAI > 150 (n = 30) or < 150 (n = 17), were recruited. CE-US showed the highest performance, with 93.5% sensitivity, 93.7% specificity, and 93.6% overall accuracy. CE-US revealed 3 bowel wall perfusion patterns after microbubble injection: submucosal enhancement and inward and outward transparietal enhancement. The linear correlation coefficient for CE-US versus CDAI was 0.74 (P < .0001); for baseline US (assessing thickness, length, and multilayer appearance of the diseased bowel) versus the CDAI, the coefficients were 0.68 (P < .0001), 0.47 (P = .0009), and 0.60 (P < .0001), respectively; and for CD-US versus CDAI the coefficient was 0.73 (P < .0001). CONCLUSIONS: CE-US has a high sensitivity and specificity in detecting inflammatory activity and a strong correlation with the CDAI.

Landscape of transcriptome variations uncovering known and novel driver events in colorectal carcinoma.

We focused on an integrated view of genomic changes in Colorectal cancer (CRC) and distant normal colon tissue (NTC) to test the effectiveness of expression profiling on identification of molecular targets. We performed transcriptome on 16 primary coupled CRC and NTC tissues. We identified pathways and networks related to pathophysiology of CRC and selected potential therapeutic targets. CRC cells have multiple ways to reprogram its transcriptome: a functional enrichment analysis in 285 genes, 25% mutated, showed that they control the major cellular processes known to promote tumorigenesis. Among the genes showing alternative splicing, cell cycle related genes were upregulated (CCND1, CDC25B, MCM2, MCM3), while genes involved in fatty acid metabolism (ACAAA2, ACADS, ACAT1, ACOX, CPT1A, HMGCS2) were downregulated. Overall 148 genes showed differential splicing identifying 17 new isoforms. Most of them are involved in the pathogenesis of CRC, although the functions of these variants remain unknown. We identified 2 in-frame fusion events, KRT19-KRT18 and EEF1A1-HSP90AB1, encoding for chemical proteins in two CRC patients. We draw a functional interactome map involving integrated multiple genomic features in CRC. Finally, we underline that two functional cell programs are prevalently deregulated and absolutely crucial to determinate and sustain CRC phenotype.

Small cell lung cancer in a young patient with osteopetrosis.

BACKGROUND: Osteopetrosis or Albers-Schönberg's disease is a heterogeneous group of rare hereditary troubles of the bone characterized by bone sclerosis due to an alteration of the bone reabsorption mediated by osteoclasts. The defect in the osteoclastic activity is responsible for complete or partial medullary cavities occlusion, with consequent reduced hemopoiesis, and for the excessive fragility of the affected bone segments. CASE REPORT: We reported the case of a young man of 31 years affected by osteopetrosis in which a small cell lung cancer developed. RESULTS: Small cell lung cancer is a particularly rare neoplasm in the young, and even though it is highly sensitive to chemotherapeutic treatment its prognosis remains poor. The greatest clinical problem connected with chemotherapeutic treatment of patients affected by osteopetrosis is the variability of the reduction of their bone marrow reserve, which could expose them to an excessive hematological toxicity caused by the therapy. CONCLUSIONS: The adoption of suitable prophylactic measures, such as the use of growth factors and drugs selected in relation to their toxicity or given in reduced doses, should be appropriately considered in these subjects.

Immunohistochemical study of androgen receptors in breast carcinoma. Evidence of their frequent expression in lobular carcinoma.

Androgens and androgen receptors (AR) are involved in the pathogenesis of breast cancer. Epidemiological studies have shown a significant association between the risk of breast cancer and androgens. However, the functional role and clinical value of AR expression in breast carcinoma have still not been clearly defined. The present study was set up to investigate the prevalence of ARs in a series of consecutive invasive breast carcinomas (IBCs) and to evaluate the patterns of AR phenotypes in a series of selected invasive lobular carcinomas (ILCs). Among the 250 consecutive IBCs (consisting of 212 ductal and 38 lobular neoplasms), AR immunoreactivity was observed in 151/250 (60.4%) cases, being expressed in 118/212 (56%) ductal and 33/38 (87%) lobular carcinomas (a statistically significant difference, chi2=11.82). AR expression was frequently associated with ER (65.2%, chi2=14.33) and PR positivity (66.9%, chi2=7.36). Most AR positive cases showed a low proliferative index (63.7%) and a low or intermediate histological grade (G1-G2, 63.9%). Among the 80 selected ILCs, AR expression was observed in 64/80 (80%) cases. Our results confirm that ARs are expressed in most breast cancers. Moreover, we demonstrated that AR positivity is particularly marked in lobular neoplasms. In addition, AR positive carcinomas are frequently characterized by a low or intermediate grade, a low proliferative index and ER and/or PR co-expression.

[Preliminary study on the expression of Toll-like receptors 7 and 8 in bladder urothelial tumors]. / Studio preliminare sull'espressione dei Toll-like receptors 7 e 8 nel tumore uroteliale della vescica.

The aim of the study was to demonstrate the expression of Toll-like receptor (TLR)-8 and to confirm the expression of TLR-7 on bladder transitional cell carcinomas (TCCs) analyzing the changes in expression between low-grade (LG) and high-grade (HG) and between non-muscle invasive (NMIBC) and muscle-invasive (MIBC) bladder cancers. In our study, 25 patients who underwent transurethral resection (TURB) for bladder TCC have been selected. Thirteen of those had an LG form,while the other cases were classified as HG; the lesions were NMIBC in 18 patients and MIBC in the others seven. The analysis of TLRs expression has been performed by immuno histochemistry. TRL-8 expression seems to be higher in MIBC (85%) than in NMIBC (78%), but it is associated with a reduced percentage of immuno reactive cells and with a lower intensity. The expression had a large nuclear localization in NMIBC (80%), althoughit was mainly cytoplasmic in MIBC (72%). TLR-7 was expressed in all NMIBC samples (where the localization was mainly nucleo-cytoplasmic) and in the 71% of MIBC samples (mainly in the nucleus). A higher expression of TRL-8 in HG TCC had been observed, while TRL-7 seems to be more expressed in LG forms. Our study allowed to document the immunohistochemical expression of TLR-8 in TCCs, confirm the immunohistochemical expression of TLR-7, and suggest an increased expression of TLR-7 in LG TCC and NMIBC, and a prevalent expression of TLR-8 in HG TCC and MIBC.

Laryngeal true malignant mixed tumor.

BACKGROUND: True malignant mixed tumor, also known as carcinosarcoma, is a salivary gland type malignant neoplasm, which is extremely rare, and only a few cases arising in the larynx have been previously reported. METHODS AND RESULTS: We report a case of true malignant mixed tumor arising in the larynx of a 65-year-old woman successfully treated with surgery. Histologically, the neoplasm was composed of variably mixed, neoplastic glandular, spindle, and chondroid tissues. Immunohistochemical analyses showed a peculiar expression of myoepithelial markers such as p63 and calponin in the glandular epithelial component, whereas malignant spindle cell proliferation was immunoreactive for calponin and actin. CONCLUSION: These results strengthen the hypothesis that these neoplasms may develop from a divergent differentiation of a totipotent, myoepithelial precursor cell. Despite the unfavorable prognosis that has always been described for these neoplasms, the patient is alive with no evidence of recurrences 5 years after surgery.

Lymph node axillary metastasis from occult contralateral infiltrating lobular carcinoma arising in accessory breast: MRI diagnosis.

We describe a case of right axillary lymph node metastasis of an occult infiltrating lobular carcinoma arising from accessory mammary gland of the left upper anterior chest wall. Ultrasonography and mammography were normal bilaterally. Magnetic resonance imaging (MRI) revealed a 3.34 cm inhomogeneous lesion. Then, core biopsy under ultrasound guidance demonstrated a typical infiltrating breast lobular carcinoma. To our knowledge, this is the first case reported in the literature of an axillary lymph node metastasis from an occult contralateral infiltrating lobular carcinoma of the accessory breast tissue. MRI was useful for assessing the lesion of the accessory breast tissue.

Interplay between Helicobacter pylori and host gene polymorphisms in inducing oxidative DNA damage in the gastric mucosa.

Infection by Helicobacter pylori is the most important risk factor for gastric cancer. However, only a small fraction of colonized individuals, representing at least half of the world's population, develop this malignancy. In order to shed light on host-microbial interactions, gastric mucosa biopsies were collected from 119 patients suffering from dyspeptic symptoms. 8-Hydroxy-2'-deoxyguanosine (8-oxo-dG) levels in the gastric mucosa were increased in carriers of H.pylori, detected either by cultural method or by polymerase chain reaction, and were further increased in subjects infected with strains positive for the cagA gene, encoding the cytotoxin-associated protein, cagA. Oxidative DNA damage was more pronounced in males, in older subjects, and in H.pylori-positive subjects suffering from gastric dysplasia. Moreover, 8-oxo-dG levels were significantly higher in a small subset of subjects having a homozygous variant allele of the 8-oxoguanosine-glycosylase 1 (OGG1) gene, encoding the enzyme removing 8-oxo-dG from DNA. Conversely, they were not significantly elevated in glutathione S-transferase M1 (GSTM1)-null subjects. Thus, both bacterial and host gene polymorphisms affect oxidative stress and DNA damage, which is believed to represent a key mechanism in the pathogenesis of gastric cancer. The interplay between bacterial and host gene polymorphisms may explain why gastric cancer only occurs in a small fraction of H.pylori-infected individuals.

Diagnostic utility of S100A1 expression in renal cell neoplasms: an immunohistochemical and quantitative RT-PCR study.

S100A1 is a calcium-binding protein, which has been recently found in renal cell neoplasms. We evaluated the diagnostic utility of immunohistochemical detection of S100A1 in 164 renal cell neoplasms. Forty-one clear cell, 32 papillary, and 51 chromophobe renal cell carcinomas, and 40 oncocytomas, 164 samples of normal renal parenchyma adjacent to the tumors and 13 fetal kidneys were analyzed. The levels of S100A1 mRNA detected by quantitative RT-PCR analysis of frozen tissues from seven clear cell, five papillary, and six chromophobe renal cell carcinomas, four oncocytomas, and nine samples of normal renal tissues adjacent to neoplasms were compared with the immunohistochemical detection of protein expression. Clear cell and papillary renal cell carcinomas showed positive reactions for S100A1 in 30 out of 41 tumors (73%) and in 30 out of 32 (94%) tumors, respectively. Thirty-seven renal oncocytomas out of 40 (93%) were positive for S100A1, whereas 48 of 51 (94%) chromophobe renal cell carcinomas were negative. S100A1 protein was detected in all samples of unaffected and fetal kidneys. S100A1 mRNA was detected by RT-PCR in all normal kidneys and renal cell neoplasms, although at very different levels. Statistical analyses comparing the different expression of S100A1 in clear cell and chromophobe renal cell carcinomas observed by immunohistochemical and RT-PCR methods showed significant values (P<0.001), such as when comparing by both techniques the different levels of S100A1 expression in chromophobe renal cell carcinomas and oncocytomas (P<0.001). Our study shows that S100A1 protein is expressed in oncocytomas, clear cell and papillary renal cell carcinomas but not in chromophobe renal cell carcinomas. Its immunodetection is potentially useful for the differential diagnosis between chromophobe renal cell carcinoma and oncocytoma. Further, S100A1 protein expression is constantly detected in the normal parenchyma of the adult and fetal kidney.