Carbonic anhydrase VI: a novel marker for salivary serous acinar differentiation and its application to discriminate acinic cell carcinoma from mammary analogue secretory carcinoma of the salivary gland.

AIMS: Carbonic anhydrase VI (CA6) is present in serous acinar cells of human salivary glands. The aim of this study was to investigate the diagnostic utility of CA6 in differentiating acinic cell carcinoma (AciCC) from its morphological mimic mammary analogue secretory carcinoma (MASC) of the salivary gland. METHODS AND RESULTS: CA6 immunostaining was performed in 28 cases of AciCC and 14 cases of MASC. All cases of AciCC showed positive CA6 staining. The staining pattern correlated with the number of serous acinar cells in tumours. All MASCs stained negatively for CA6. The results were compared with those obtained with currently used markers, including DOG1, mammaglobin, S100, and vimentin. CA6 showed sensitivity and specificity as high as those of DOG1 in diagnosing AciCC. CA6 expression was focally observed in basal cell adenoma and in one case of cystadenocarcinoma (1/3), but not in other salivary gland tumours, including mucoepidermoid carcinoma, adenoid cystic carcinoma, salivary duct carcinoma, lymphoepithelial carcinoma, epithelial-myoepithelial carcinoma, and pleomorphic adenoma. CONCLUSIONS: CA6 is a specific marker for serous acinar cells of salivary glands and AciCC. CA6 has sensitivity and specificity equal to those of DOG1 in the differential diagnosis between AciCC and MASC. A combination of CA6 and DOG1 could be an ideal immunohistochemical panel for AciCC.

Expression of LEF1 is an independent prognostic factor for patients with oral squamous cell carcinoma.

BACKGROUND/PURPOSE: Lymphoid-enhancing factor 1 (LEF1) is a transcription factor mediating Wnt/ß-catenin signaling. In this study, we analyzed the clinicopathologic significance of LEF1 expression in oral squamous cell carcinoma (OSCC). METHODS: Expression levels of LEF1 in 135 cases of OSCC were determined by immunohistochemistry. The results were correlated with clinicopathologic parameters and patient outcome. RESULTS: LEF1 was only occasionally detected in basal and parabasal cells of nontumorous squamous epithelium. Overexpression of LEF1 was observed in 33 of 135 OSCCs (24%). LEF1 was more frequently expressed in moderately to poorly differentiated cancer (p = 0.0035) and was associated with lymphovascular invasion (p = 0.0252). Overexpression of LEF1 was significantly associated with poor prognosis (p = 0.0176, hazard ratio = 1.96, 95% CI = 1.02-3.75). Multivariate analysis revealed LEF1expression and margin status to be the significant independent predictors for overall survival. CONCLUSION: Our study suggests LEF1 expression in OSCC may play an important role in tumor progression and can be served as a predictor of poor prognosis for patients with OSCC.

Insulin-like growth factor II mRNA-binding protein 3 expression promotes tumor formation and invasion and predicts poor prognosis in oral squamous cell carcinoma.

BACKGROUND: Insulin-like growth factor II mRNA-binding protein 3 (IGF2BP3), an oncofetal RNA-binding protein, has been implicated in the enhancement of proliferation and invasion in various cancers. This study aimed to investigate the clinical significance and functional role of IGF2BP3 expression in oral squamous cell carcinoma (OSCC). METHODS: IGF2BP3 expression in 93 OSCC patients was investigated using immunohistochemical staining and correlated with clinical parameters and patients' survival. The effect of IGF2BP3 on cell invasion ability was evaluated by RNA interference in OSCC cell line. RESULTS: High expression of IGF2BP3 in OSCC was significantly correlated with large tumor size and lymph node metastasis. Kaplan-Meier analysis revealed that oral cancer patients with high IGF2BP3 expression had a significantly lower 5-year survival (P = 0.0017). Multivariate analysis of clinical samples demonstrated IGF2BP3 to be an independent prognosis factor (P = 0.003). Moreover, the IGF2BP3 shRNA significantly suppressed the invasion ability of OSCC in vitro, and the knockdown of endogenous IGF2BP3 expression also inhibited tumor formation in vivo. CONCLUSIONS: IGF2BP3 enhances cell invasion ability and tumorigenicity in human OSCC in vitro and in vivo. IGF2BP3 is an independent prognostic factor in patients with OSCC. Targeting of IGF2BP3 could potentially suppress the tumor growth and metastasis to improve the outcome of patients with OSCC.

Cadherin-17 is a useful diagnostic marker for adenocarcinomas of the digestive system.

Cadherin-17, also called liver-intestine cadherin, is a calcium-dependent transmembrane glycoprotein that mediates cell-cell adhesion in intestinal epithelium. Expression of cadherin-17 was reported in gastric, pancreatic, and colorectal adenocarcinomas but not in other tumors. Whether cadherin-17 can be used as a marker for diagnosis of cancers is still unclear. In this study, we used immunohistochemical methods to stain cadherin-17 in tissue arrays containing most normal tissues and 518 carcinomas from many anatomic sites. Among normal tissues, the expression of cadherin-17 was limited to epithelial cells of small intestine and colon. Colorectal adenocarcinomas showed staining in 96% of cases and most of them had strong and diffuse staining. Gastric, pancreatic, and biliary adenocarcinomas showed diffuse or scattered staining in about 25-50% of cases. Fewer than 1% of carcinomas outside the digestive system were positive for cadherin-17. When a two-marker, Cadherin-17/cytokeratin 7, profile was used, 37 of 38 (97%) cadherin-17(+)/cytokeratin 7(-) tumors were colorectal adenocarcinomas; 49 of 56 (86%) cadherin-17(+)/cytokeratin 7(+) tumors were gastric, pancreatic, or biliary adenocarcinomas. Our results show that cadherin-17 is a useful immunohistochemical marker for diagnosis of adenocarcinomas of the digestive system.

Label-free imaging of Drosophila larva by multiphoton autofluorescence and second harmonic generation microscopy.

The fruit fly Drosophila melanogaster is one of the most valuable organisms in studying genetics and developmental biology. To gain insight into Drosophila development, we successfully acquired label-free, in vivo images of both developing muscles and internal organs in a stage 2 larva using the minimally invasive imaging modality of multiphoton autofluorescence (MAF) and second harmonic generation (SHG) microscopy. We found that although MAF is useful in identifying structures such as the digestive system, trachea, and intestinal track, it is the SHG signal that allowed the investigation of the muscular architecture within the developing larva. Our results suggest that multiphoton microscopy is a powerful in vivo, label-free imaging technique to examine Drosophila physiology and may be used for developmental studies.

Prognostic role of c-Jun activation in patients with areca quid chewing-related oral squamous cell carcinomas in Taiwan.

BACKGROUND: Overexpression or activation of c-Jun has been implicated in the pathogenesis of several types of cancer. Treatment of oral cells with areca nut extract in culture can increase c-Jun proto-oncogene mRNA levels. The purpose of this study was to investigate the possible role of c-Jun activation in the pathogenesis and prognosis of areca quid chewing-related oral cancer in Taiwan. METHODS: We immunohistochemically examined c-Jun protein activation in human oral squamous cell carcinomas (SCC) using an anti-phosphospecific c-Jun (pc-Jun) antibody on paraffin-embedded sections. RESULTS: Positive pc-Jun staining was observed in 42 of 70 (60%) cases of oral SCC. No significant correlation was found between pc-Jun expression and patients' age, gender, oral habit, cancer location, clinical stage, tumor size and lymph node status. Kaplan-Meier curves showed that oral SCC patients with positive pc-Jun staining or positive lymph node metastasis had significantly shorter overall survival (p < 0.018 and 0.001, respectively, by log-rank test). CONCLUSION: These results indicate that c-Jun activation may play an important role in the carcinogenesis of oral SCC. Positive pc-Jun staining may serve as an adjuvant marker of worse prognosis in patients with oral SCC in Taiwan.

Survivin expression predicts poorer prognosis in patients with areca quid chewing-related oral squamous cell carcinoma in Taiwan.

Survivin, a recently characterized novel member of the inhibitor of apoptosis (IAP) family, is not detectable in most differentiated normal adult tissues but is expressed in a wide range of cancer tissues. Its expression in cancer has been correlated with poor prognosis, cancer progression and drug resistance. We immunohistochemically examined the expression of survivin in 62 cases of oral epithelial dysplasia (ED) and 96 cases of oral squamous cell carcinoma (SCC). Cytoplasmic survivin staining was detected in 60 of the 62 (97%) ED specimens and 94 of the 96 (98%) SCC specimens but not in adjacent normal oral mucosal tissues. The labeling index (LI) for survivin protein significantly increased from ED (32.3+/-16.3%) to SCC samples (49.4+/-28.5%) (p<0.001). In addition, the mean LI for ED cases with further malignant transformation into SCC (45.6+/-8.8%) was higher than those without malignant transformation (30.1+/-16.3%) (p=0.008). No significant correlation was found between the survivin expression and the patients' age, sex, oral habit, cancer location, or STNM status in SCC cases. Kaplan-Meier curves showed oral SCC patients with high survivin expression (LI>25%), advanced stage, larger tumor size, or positive lymph node metastasis had significantly shorter overall survival (p=0.014, 0.012, 0.005 and 0.011, respectively by log-rank test) than others. The associations remained significant after adjusting for age. These results indicate that survivin protein expression may be an important early event in oral carcinogenesis and predicts unfavorable prognosis for oral SCC. Furthermore, the unique expression of survivin in cancer cells but not in most normal adult tissues suggests that modulation of survivin protein expression may provide a novel strategy for the therapy of oral SCC.

Overexpression and amplification of Aurora-A in hepatocellular carcinoma.

PURPOSE: Aurora-A/STK15/BTAK, a centrosome-associated serine/threonine kinase, has been shown to induce chromosomal instability, leading to aneuploidy and cell transformation. The purpose of this study was to investigate the expression and amplification of Aurora-A in hepatocellular carcinoma (HCC). EXPERIMENTAL DESIGN: Aurora-A mRNA levels were measured in 224 HCCs and 199 paired nontumorous liver tissues by reverse transcription-PCR. Aurora-A mRNA and protein levels of 8 were also measured by reverse transcription-PCR and Western blot hybridization in 8 liver cancer cell lines. Amplification of Aurora-A was determined by Southern blot hybridization in 99 cases. RESULTS: Aurora-A was overexpressed in 137 of 224 (61%) HCCs and all 8 of the cell lines. Overexpression of Aurora-A was associated with high-grade (grade II-IV), and high-stage (stage IIIB-IV) tumors, p53 mutation, infrequent beta-catenin mutation, and poor outcome. Aurora-A overexpression and p53 mutation acted synergistically toward poor prognosis. Amplification of Aurora-A was detected only in 3 HCCs. CONCLUSION: The results show that Aurora-A is overexpressed frequently in HCC, and correlated with high grade and high stage, indicating that overexpression of Aurora-A plays a role in the development and progression of HCC.

Expression of cyclin A is related to progression of oral squamous cell carcinoma in Taiwan.

Cyclin A is required for DNA synthesis during the S phase and progression through the G2/M transition. Increased expression of cyclin A protein has been correlated with poor prognosis in a variety of human tumors. To investigate the possible influence(s) of cyclin A protein on the progression and prognosis of oral squamous cell carcinomas (SCCs) in Taiwan. We examined the expression of cyclin A in oral SCC, epithelial dysplasia (ED) and normal oral mucosa (NOM) by immunohistochemistry using antibodies to cyclin A. Results and Conclusions. The mean labeling indices (LI) in NOM, ED and SCCs were 7.0+/-3.1%, 12.1+/-3.9% and 21.3+/-12.3%, respectively. The cyclin A LI for oral SCCs was significantly higher than that for NOM (P=0.002) or ED (P<0.001). In addition, a high LI for cyclin A was found to correlate with advanced stage (P=0.0048), larger tumor size (P=0.0017), lymph node involvement (P=0.0006) and cancer recurrence (P<0.0001). The Kaplan-Meier analysis showed patients with tumors containing more than 15% cyclin A-positive cells had significantly shorter overall survival than those with tumors containing less than 15% cyclin A-positive cells (P<0.00001). These results indicate that overexpression of cyclin A protein is associated with tumor progression and patient prognosis for oral SCC.

Expression of Cyr61 (CCN1) in human oral squamous cell carcinoma: An independent marker for poor prognosis.

BACKGROUND: Cysteine-rich 61 (Cyr61 [CCN1]) has disparate functions in tumorigenesis that are dependent on the cell types. The aim of the study was to investigate its role in the growth of oral squamous cell carcinoma (SCC). METHODS: The study used immunohistochemistry to examine Cyr61 expression in 93 oral SCC specimens and assessed the effect of Cyr61 overexpression on proliferation and migration of oral SCC cells in vitro and xenograft growth in severe combined immunodeficient (SCID) mice. RESULTS: High expression of Cyr61 significantly correlated with large tumor size (p = .009) and advanced tumor stage (p = .036). Multivariate analysis revealed that high Cyr61 (relative risk [RR] 2.44, 95% confidence interval [CI] 1.209-4.95, p = .010) significantly correlated with mortality. Forced expression of Cyr61 stimulated the motility and growth of Ca9-22 cells in vitro and enhanced xenograft growth in SCID mice. CONCLUSIONS: Cyr61 is a positive growth modulator of oral SCC and Cyr61 overexpression is an independent prognostic indicator for patients with oral SCC.

Histone deacetylase 2 expression predicts poorer prognosis in oral cancer patients.

Histone deacetylase 2 (HDAC2) has been implicated in the development and progression of several human tumors. We immunohistochemically examined the expression of HDAC2 protein in 20 cases of oral epithelial dysplasia (OED) and 93 cases of oral squamous cell carcinoma (OSCC). Positive HDAC2 nuclear staining was observed in 80 of the 93 (86.02%) cases of SCC and 11 of the 20 (55%) cases of ED. The labeling index (LI) for HDAC2 nuclear staining increased significantly from ED (25.8+/-26.5%) to SCCs (59.8+/-28.5%) (p<0.001). No significant correlation was found between the HDAC2 expression level and patient's age, sex, oral habits in oral SCC patients. However, cancer with advanced stage, larger tumor size, or positive lymph node metastasis had higher level of HDAC2 protein expression. Kaplan-Meier curves showed oral SCC patients with high HDAC2 expression (LI>50%), advanced stage, larger tumor size, or positive lymph node metastasis had significantly shorter overall survival (p=0.0158, 0.0267, 0.0029 and 0.02514, respectively by log-rank test) than others. The results of this study show for the first time that overexpression of the HDAC protein is a frequent event in oral cancer and could be used as a prognostic factor in oral SCC.

Nuclear localization of annexin A1 is a prognostic factor in oral squamous cell carcinoma.

BACKGROUND AND OBJECTIVES: To investigate whether annexin A1 (ANXA1) expression is a marker in predicting the prognosis of oral cancer patients. METHODS: We immunohistochemically examined the expression of ANXA1 in 66 cases of oral epithelial dysplasia (OED) and 115 cases of oral squamous cell carcinoma (OSCC). The results were correlated with the clinicopathological parameters of tumors and overall patient survival. RESULTS: In normal oral mucosa, ANXA1 staining was predominantly located on the cell membrane. In OED and OSCC specimens, membranous staining decreased, whereas nuclear staining increased. Positive nuclear staining was observed in 9 of 66 (13.64%) OED cases and 63 of 115 (54.8%) OSCCs. Kaplan-Meier curves showed that OSCC patients with ANXA1 nuclear staining had significantly shorter overall lengths of survival (P = 0.00036 by the log-rank test). Multivariate analysis showed that ANXA1 nuclear staining is a significant predictor of poor overall survival. And oral cancer SAS cells treated with hepatocyte growth factor (HGF) can induce ANXA1 protein translocation from cytoplasm to nucleus. Cells pretreated with LY294002 (PI3K inhibitor) almost completely inhibited (88.3% inhibition) HGF-mediated ANXA1 nuclear translocation. CONCLUSIONS: The nuclear localization of ANXA1 protein is a frequent event and could be used as a prognostic factor in OSCC.

Down-regulation of annexin A10 in hepatocellular carcinoma is associated with vascular invasion, early recurrence, and poor prognosis in synergy with p53 mutation.

Annexins (ANXs) are a large group of calcium-binding proteins participating in diverse important biological processes. ANXA10 is the least expressed new member of unknown function. We showed that ANXA10 mRNA was expressed in adult liver and hepatocellular carcinoma (HCC), but not in multiple adult and fetal tissues, cholangiocarcinoma, and several other common carcinomas. Of 182 unifocal primary HCCs, ANXA10 mRNA was dramatically reduced in 121 (66%), and the down-regulation correlated with p53 mutation (P = 0.024), early intrahepatic tumor recurrence (P = 0.0007), and lower 4-year survival (P = 0.0014). Down-regulation of ANXA10 was twofold more frequent in large than small HCCs (P = 0.0012), in grade II to III than grade I HCC (P < 0.00001), and in stage IIIA to IV than stage I to II HCC (P < 0.00001). Moreover, ANXA10 down-regulation and p53 mutation acted synergistically toward high-grade (P < 0.00001), high-stage HCC (P < 0.00001), and poorer prognosis (P = 0.0025). Our results indicate that the expression of the tissue- and tumor-restricted ANXA10 is a marker of liver cell differentiation and growth arrest, and its down-regulation associated with malignant phenotype of hepatocytes, vascular invasion, and progression of HCC, leading to poor prognosis. Thus, ANXA10 might serve as a new potential target of gene therapy for HCC.