Overexpression of Lox in triple-negative breast cancer

BackgroundTriple negative breast cancer (TNBC) accounts for approximately 15% of breast cancers. It is associated with a poor prognosis and typically earlier onset of metastasis in comparison with other breast cancer subtypes. Since TNBC lacks the expression of estrogen and progesterone receptors and Her2 status is also negative, there is currently no target that can be used for systemic therapy. Epithelial-mesenchymal transition (EMT) plays an important role in tumor progression and metastasis. In this study, we examined a subset of EMT markers consisting of Snail, Twist-1 and Lox in TNBC and non-TNBC breast cancer subtypes and analyzed their expression pattern in regard to subtype, clinico-pathological parameters and prognosis.Experimental designWe analyzed 659 breast cancer samples from two tissue microarrays. Breast cancer samples were categorized into two groups according to hormone receptor expression and Her2 status (n = 146 were triple negative, n = 513 were non triple-negative). Immunohistochemical expression of Snail, Twist-1 and Lox was semi-quantitatively analyzed using a three-tiered (weak-moderate-strong) scoring system. Results were statistically analyzed and correlated to clinico-pathological parameters and overall survival.ResultsStrong overexpression of Lox was significantly higher in triple negative breast cancers when compared to non triple-negative breast cancers (p < 0.001). No difference was seen between the groups regarding Snail and Twist expression (p > 0.05). In addition, Lox expression was significantly stronger in poorly differentiated (G3) breast cancers (p < 0.001 for Lox).ConclusionsThe EMT marker Lox has a differential expression pattern in breast cancer, being significantly overexpressed in triple negative breast cancers. We could not link this expression to prognosis, however, this marker might be explored in future studies as possible target for systemic therapy of TNBC.     

Predictive and prognostic values of BubR1 and synuclein-gamma expression in breast cancer

The aim of this study is to determine the expression level of spindle assembly checkpoint (SAC) proteins-BubR1 and synuclein-gamma (SNCG) in human breast cancer tissues and to test whether there is a relationship between their expression levels and clinicopathologic parameters including respons to taxanes, tumor grade, estrogen receptor (ER) pozitivity, HER2 status, and overall survival (OS). We analyzed retrospectively paraffin-embedded tissue sections from 55 breast cancer patients whose clinical outcomes had been tracked after taxane treatment in neoadjuvan and metastatic setting. The expression status of BubR1 and SNCG was defined by immunohistochemistry (IHC) using the anti-BubR1 and anti-SNCG antibody. The BubR1 and SNCG was overexpressed in 38% and 62% of the study group, respectively. There was borderline significant correlation between low BubR1 expression and increased taxane sensitivity (P=0.05). In contrast, high SNCG expression was significantly associated with decreased taxane sensitivity (P=0.01). There was no association between the clinicopathologic parameters including histologic grade, ER positivity and HER2 status and the level of these proteins. However, triple negative tumors showed significantly more high BubR1 expression than those other molecular subtypes (P=0.04). Kaplan-Meier survival analysis failed to show a significant correlation between expression levels of BubR1 and SNCG and overall survival although patients with low levels of both proteins had a marginally longer survival time compared to those with high levels. In summary, our data suggest that both BubR1 and SNCG may be promising predictive marker rather than prognostic marker in patients with breast cancer.     

Decreased expression of histone deacetylase 10 predicts poor prognosis of gastric cancer patients

Aberrant expression of histone deacetylase (HDACs) was associated with carcinogenesis and progression of various tumors. However, the association of HDAC10 with clinical outcomes in gastric cancer patients is unclear. Thus, the objective of the current study was to evaluate the association of expression level of HDAC10 with clinicopathologic factors and prognosis of patients with gastric cancer. The expression level of HDAC10 in 179 paraffin-embedded gastric cancer tissue specimens was examined by immunohistochemistry (IHC). As a result, we found that expression of HDAC10 in gastric cancer was significantly decreased in gastric cancer tissues as compared with adjacent tissues (51.4% vs. 87.3%, P < 0.001). HDAC10 expression was significantly correlated with gender (P = 0.023), tumor size (P = 0.015), histological grade (P = 0.009), tumor invasion (P = 0.033), lymph node metastatic status (P = 0.019) and tumor stage (P = 0.004), but not correlated with age and lauren classification (all P > 0.05). Kaplan-Meier survival curves showed that the overall survival rate was significantly lower in the patients with low expression of HDAC10 compared with those patients with high HDAC10 (P < 0.001). Moreover, multivariate analysis revealed that HDAC10 expression was an independent prognostic factor for gastric cancer patients (P = 0.001). These results suggest that HDAC10 expression could see as a prognosis marker for gastric cancer patients.     

Lemur tyrosine kinase-3 is a significant prognostic marker for patients with colorectal cancer

Lemur tyrosine kinase-3 (LMTK3) belongs to the family of serine-threonine-tyrosine kinases and the aberrant expression of LMTK3 was observed in several human malignancies. However, the association of LMTK3 with clinical outcomes in colorectal cancer patients is unclear. Thus, this present study was to evaluate the association of LMTK3 expression level with clinicopathologic factors and prognosis of patients with colorectal cancer (CRC). The expression level of LMTK3 in 69 archival paraffin-embedded colorectal tumor tissue specimens was examined by immunohistochemistry (IHC). As a result, we found that the LMTK3 expression level was significantly elevated in CRC tissues as compared with Crohn’s disease or colorectal polyp tissues (P<0.0001, P<0.0001, respectively). Positive LMTK3 signals in the colorectal cancer cells were observed in about 89.9% (62 of 69) CRC tissue specimens. Additionally, LMTK3 expression was significantly correlated with lymph node metastasis and tumor-node-metastasis (TNM) classification (P=0.003, and P=0.008, respectively), but not with sex, age, tumor location, histological differentiation, tumor size, or depth of tumor invasion (all P>0.05). Kaplan-Meier survival curves showed that the overall survival rate was significantly higher in the patients with low expression of LMTK3 when compared with those patients with high LMTK3 (P=0.010). Moreover, multivariate analysis revealed that LMTK3 expression was an independent prognostic factor for CRC patients (P=0.047). These results suggest that LMTK3 protein could serve as a prognostic marker for CRC patients.     

Low PBRM1 identifies tumor progression and poor prognosis in breast cancer

Background: To investigate the expression and role of PBRM1 in breast cancer, and to evaluate the clinical and prognostic significance of PBRM1 protein in patients with breast cancer. Methods: The expression of PBRM1 was examined in breast cancer tissue and paired non-cancerous tissues by real-time PCR. Moreover, PBRM1 protein expression was evaluated by immunohistochemistry in 150 paraffin-embedded breast cancer specimens. The correlation between PBRM1 expression and clinicopathological features were statistically analyzed. Results: The status of PBRM1 protein in breast cancer tissues is much lower than that in paracarcinoma tissues. Low PBRM1 expression was positively correlated with tumor stage (P =0.003) and lymph node metastasis (P =0.013). The overall (P =0.003) and recurrent-free survival (P =0.001) of the patients with high PBRM1 expression was significantly lower than the low PBRM1 expression group. Multivariate analysis showed that the expression of PBRM1 was an independent factor of overall survival for the patients with breast cancer (P =0.030). Conclusions: PBRM1 might involve in the development and progression of breast cancer as a tumor suppressor, and thereby may be a valuable prognostic marker for breast cancer patients.     

Immunohistochemical evaluation of human epidermal growth factor receptor 2 and estrogen and progesterone receptors in invasive breast cancer in women

Introduction

Estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER-2) expression are crucial in the biology of breast carcinoma. HER-2/neu gene is amplified and overexpressed in 15-30% of invasive breast cancers. HER-2-positive breast cancers have worse prognosis than HER-2 negative tumors and possess distinctive clinical features. The aim of this study was to assess the expression of HER2 in cancer tissue of patients with invasive breast cancer in correlation with tumor type, histological grade, tumor size, lymph node status, and expression of estrogen receptor and progesterone receptor.

Material and methods

Formalin-fixed, paraffin-embedded tissues from 40 patients with invasive HER-2-positive breast cancer and from 191 patients with HER-2-negative breast cancer were used in this study. HER2 expression was determined using the test HerceptTest™ DAKO.

Results

Among 231 cases of breast cancer, 18 invasive lobular carcinomas and 213 invasive ductal carcinomas were diagnosed. Sixty percent of HER-2-positive breast cancers were ER-positive compared with 77% in the HER-2-negative group (p = 0.002). The expression of PR was observed in 43% of HER-2-positive breast cancers and in 72% of HER2-negative tumors (p = 0.003). Excessive expression of HER2 protein was detected in 60% of patients positive for estrogen receptors, which may worsen prognosis in these patients.

Conclusions

Determination of HER2 overexpression in breast cancer patients, allows for a determination of a group of patients with a worse prognosis.     

Prognostic significance of HOXD13 expression in human breast cancer

Homeobox protein Hox-D13 has been recognized as a tumor suppressor in pancreatic cancer. To evaluate the function of HOXD13 in invasive breast cancer pathogenesis, we examined HOXD13 expression in 434 breast cancer tissues and 230 their counterpart normal breast tissues by immunohistochemistry using a tissue microarray (TMA). The association between HOXD13 expression and clinicopathological factors was analyzed by use of Chi-square test. Kaplan-Meier survival curves and log-rank tests were applied to analyze the survival status. Cox regression was applied for multivariate analysis of prognosis. We found that low HOXD13 expression accounts for 84.3% in breast cancer tissues. Low HOXD13 expression was significantly associated with large tumor size (P=0.038) and positive lymph node metastasis (LNM) (P=0.026). In Kaplan-Meier survival curves and log-rank tests, the patients with HOXD13-negative breast cancer showed significantly poorer outcomes (69.867 ± 1.058 months) in terms of overall survival (OS) than positive-HOXD13-expression patients (76.248 ± 1.069 months) (P=0.003). And in multivariate analysis, low level of HOXD13 expression was a significant unfavorable prognostic factor. So we conclude that down-regulation of HOXD13 might be a potentially useful prognostic marker for patients with breast cancer.     

Association between carbonic anhydrase 9 expression and poor prognosis in sinonasal squamous cell carcinoma

ObjectivesCarbonic anhydrase 9 (CA9), as a member of the carbonic anhydrase enzyme family, was an endogenous marker of hypoxia. Previous studies suggested CA9 expression was correlated with poor prognosis in multiple types of malignancies. Therefore, this study was to evaluate the role of CA9 in sinonasal squamous cell carcinoma (SNSCC) and to determine whether this biomarker was associated with patient clinicopathologic characteristics and prognosis.MethodsWe assessed 63 patients diagnosed with SNSCC in 2013–2017 who underwent curative surgery. Tumor specimens was immunohistochemically analyzed for CA9 expression. The expression levels of CA9 was evaluated in relation to clinicopathological factors and prognosis.ResultsPositive expression of CA9 was observed in 21 (33.3%) patients and was significantly correlated with local recurrence (p = 0.016), overall survival (OS) (p = 0.003) and disease-free survival (DFS) (p = 0.002). In Cox's multivariate analysis, CA9 expression was an independent negative prognostic factor for OS (p = 0.048) and DFS (p = 0.019).ConclusionsOur findings demonstrated that CA9 overexpression could be used as an independent prognostic biomarker and therapeutic target in SNSCC.     

High expression of flotillin-2 is associated with poor clinical survival in cervical carcinoma

Aims: To investigate the expression and clinical significance of flotillin-2 (FLOT2) in cervical cancer (CC). Methods: We examined FLOT2 mRNA levels in 10 pairs of cervical cancer and adjacent normal tissues. Immunohistochemistry was performed to analyze FLOT2 protein expression in 115 archived cervical cancer samples. The association between FLOT2 levels, clinicopathologic factors and prognosis was analyzed statistically as well. Results: The cancer tissues of CC patients had clearly increased expression of FLOT2 at mRNA level as compared to adjacent nontumorous tissues. Survival analysis of CC patients indicated that FLOT2 expression was significantly associated with poor overall and local recurrence-free survival (P = 0.025 and P = 0.028, respectively). Moreover, FLOT2 expression was significantly correlated with clinical stage, tumor differentiation, and lymph nodes metastasis. Multivariate analysis revealed that FLOT2 expression was an independent prognostic factor for overall survival in CC patients. Conclusion: FLOT2 may serve as an oncogene in the development of CC, and may serve as a clinicopathologic biomarker for prognosis in CC patients.     

LncRNA PCAT6: A potential biomarker for diagnosis and prognosis of bladder cancer

BackgroundMany potential biomarkers have been identified and studied for bladder cancer diagnosis. In this study, we investigated the role of a new biomarker, long noncoding RNA (lncRNA) PCAT6, in bladder cancer diagnosis and prognosis.Methods and resultsThe lncRNA PCAT6 expression profile of BC is analyzed using the Cancer Genome Atlas Urothelial Bladder Carcinoma (TCGA-BLCA) data. PCAT6 expression level in 106 pairs of BC tissues and adjacent normal tissues was detected and compared using qRT-PCR. Then, the association between PCAT6 expression and clinicopathologic indicators of BC was evaluated. Meanwhile, the prognostic value of PCAT6 was tested using Kaplan-Meier analysis. Additionally, loss-of-function assays were used to explore the effect of PCAT6 on the biological function of BC cells.We identified that the expression level of PCAT6 in BC tissue was higher than that in adjacent normal tissues. And the BC patients have higher serum PCAT6 than that in healthy volunteers. In addition, the expression level of PCAT6 was correlated with tumor size (p = 0.005), differentiation (p = 0.018), TNM stage (p = 0.04), lymph nodes metastasis (p = 0.019), and distant metastasis (p = 0.028). Kaplan-Meier analysis showed that BC patients with high PCAT6 expression had shorter overall survival (OS) and progression-free survival (PFS). The loss-of-function results revealed that the proliferation and viability of BC cells in PCAT6 knockdown groups decreased significantly, compared with the negative control groups.ConclusionOur results demonstrated that PCAT6 might be a potential biomarker for diagnosis and prognosis of BC.     

Overexpression of nucleolin and different expression sites both related to the prognosis of gastric cancer

The aim of this study was to investigate the expression of nucleolin in tumorous tissues and corresponding non‐malignant tissues in gastric cancer (GC), and the correlation of different expression sites with clinicopathologic parameters and prognosis. Immunohistochemistry was used for detecting the expression levels of nucleolin in GC tissues and corresponding non‐malignant tissues from 124 gastrectomy specimens with stage I–III. Staining results were correlated with clinicopathologic features and survival. Both GC tissues and corresponding non‐malignant tissues showed nucleolar staining for nucleolin. Nucleolin expression was higher in GC tissues than in non‐malignant tissues. Among the 124 GCs, 85 (68.5%) were nucleolin‐high. No significant correlation between nucleolin expression and other clinicopathologic parameters was found. The Cox univariate analysis indicated that both cytoplasmic staining and nucleolar staining of nucleolin expression correlated with patients' prognosis (log‐rank, p < 0.0001; p = 0.0075, respectively). It was concluded in the study that nucleolin was overexpressed in GCs. A high level of nucleolar expression of nucleolin was an independent prognostic marker for better survival while high cytoplasmic staining was closely associated with worse prognosis for GC patients.     

Immunohistochemical expression and prognostic relevance of Bmi-1, a stem cell factor,in epithelial ovarian cancer

Ovarian cancer is the fourth most common cause of cancer-related death in women. Bmi-1 is a stem cell factor implicated in many human malignancies with poor outcome. Few published reports on the expression of Bmi-1 in epithelial ovarian cancer were either experimental or performed on cell lines. This study evaluates the immunohistochemical expression of Bmi-1 protein in epithelial ovarian cancer tissue specimens and its relevance to the clinicopathologic prognostic variables and patient survival. Forty cases of epithelial ovarian cancer were selected according to the availability of paraffin-embedded tissue and the clinicopathologic and survival data. Immunohistochemistry was performed for anti–Bmi-1 antibody. Low and high Bmi-1 expression groups were compared with age, tumor stage, laterality, grade, histology, and patient survival. Bmi-1 expression was detected in 72.5% of cases, of which 42.5% had high expression. High Bmi-1 expression strongly associated with advanced International Federation of Gynecology and Obstetrics stages (P = .007), bilaterality (P = .01), and higher Gynecologic Oncology Group grades (P = .031) and carcinomas of serous histology (P = .027). It had no association with patient age. Bmi-1 expression displayed a significant inverse association with patient overall and mean survival (P = .006, P < .001). These observations suggested correlation between increased Bmi-1 expression and clinical progression in ovarian epithelial cancer.     

Significant down-regulation of growth hormone receptor expression revealed as a new unfavorable prognostic factor in hepatitis C virus-related hepatocellular carcinoma

Background/AimsGrowth hormone (GH) is the main regulator of somatic growth, metabolism, and gender dimorphism in the liver. GH receptor (GHR) signaling in cancer is derived from a large body of evidence, although the GHR signaling pathway involved in the prognosis of hepatocellular carcinoma (HCC) in patients with hepatitis C virus (HCV)-related HCC, remains unclear. We aimed to explore the expression of GHR and analyze its association with clinicopathologic features and prognosis of patients with chronic hepatitis C and HCC.MethodsThe expression of GHR mRNA was investigated by quantitative real-time polymerase chain reaction in paired tumors and adjacent non-tumorous (ANT) liver tissues of 200 patients with chronic hepatitis C and HCC. Western blotting and immunofluorescence assays using the HCV-infected Huh7.5.1 cell model was performed.ResultsGHR mRNA was significantly lower in HCV-HCC tissues than in corresponding ANT liver tissues. GHR mRNA and protein levels also decreased in the HCV-infected Huh7.5.1 cell model. Notably, lower GHR expression was associated with age of >60 years (P=0.0111) and worse clinicopathologic characteristics, including alpha-fetoprotein >100 ng/mL (P=0.0403), cirrhosis (P=0.0075), vascular invasion (P=0.0052), pathological stage II–IV (P=0.0002), and albumin ≤4.0 g/dL (P=0.0055), which were linked with poor prognosis of HCC. Most importantly, the high incidence of recurrence and poor survival rates in patients with a low ratio of tumor/ANT GHR (≤0.1) were observed, indicating that low expression levels of GHR had great risk for development of HCC in patients with chronic hepatitis C.ConclusionsOur study demonstrates a significant down-regulation of GHR expression as a new unfavorable independent prognostic factor in patients with chronic hepatitis C and HCC.     

Overexpression of Arginase-1 is an indicator of poor prognosis in patients with colorectal cancer

AimArginase-1 (Arg-1) metabolizes l-arginine to l-ornithine and urea. It has been documented to have a role in various malignancies. However, the relationship between Arg-1 expression and clinicopathological characteristics of colorectal cancer (CRC) patients remains to be elucidated. The present study aimed to analyze the expression and prognostic value of Arg-1 in patients with CRC.Material and methodsThe mRNA and protein expressions of Arg-1 in fresh colorectal cancer tissue specimens and the corresponding noncancerous tissue specimens were examined by RT-qPCR (n = 24) and western blot analysis (n = 17). Arg-1 expression levels were determined in paraffin-embedded CRC tissue specimens (n = 236) by immunohistochemistry. The associations of Arg-1 expression and clinicopathological features and clinical prognosis in 236 CRC patients were analyzed.ResultsThe expression levels of Arg-1 were significantly higher in the CRC tissues compared with the matched noncancerous tissues, and elevated Arg-1 expression was remarkably associated with stage III-IV tumors (P = 0.007), lymph node metastasis (P = 0.019) and a plasma albumin concentration <35 g/l (P = 0.022). Kaplan-Meier analysis indicated that Arg-1 overexpression was associated with adverse prognoses for overall survival (OS) (P < 0.001) and disease-free survival (DFS) (P < 0.001) in all cases. Further analysis revealed that the patients with high Arg-1 expression had significantly shorter OS and DFS at the advanced stages (III + IV) (P = 0.032 for OS, and P = 0.012 for DFS) but not at the early stages (I + II) (P = 0.194 for OS, and P = 0.065 for DFS). Multivariate analysis revealed that Arg-1 overexpression was an independent prognostic factor for OS (P = 0.002) and DFS (P < 0.001) in patients with CRC.ConclusionThe data indicated that Arg-1 overexpression in CRC may be a marker that can discriminate subgroups of patients with a poor prognosis.     

HPD overexpression predicts poor prognosis in breast cancer

BackgroundThe enzyme, 4-hydroxyphenylpyruvate dioxygenase (HPD), is critical to tyrosine metabolism; its deficiency can cause tyrosinemia. However, its precise contribution to tumorigenesis is unclear. Here, we investigated the correlation between HPD expression and prognosis in patients with breast cancer.Methods145 breast cancer specimens were selected to analyze HPD protein expression by immunohistochemistry and evaluate its relationship to patients’ clinicopathological features. HPD localization was confirmed in MCF-7 and MDA-MB-231 breast cancer cells, using immunofluorescence staining. The expression of HPD protein was detected in breast cancer and cancer-adjacent normal tissues using Western blot analysis. Survival rates were calculated by the Kaplan–Meier method.ResultsWe found that HPD protein was mainly located in the cytoplasm/nucleoli/perinucleus in breast cancer cells, as shown by immunofluorescence staining in MCF-7 and MDA-MB-231 cells, and immunohistochemistry in breast cancer and adjacent normal tissues (HPD protein expression—breast cancer: 46.9% [68/145], ductal carcinoma in situ [DCIS]: 22.6% [12/53], and normal tissues: only 4.8% [2/42]). Similarly, the Western blot results further confirmed the increased expression of HPD in breast cancer compared with cancer-adjacent normal tissues (P < 0.05). HPD expression level was positively correlated with histological grade and clinical stage, and inversely correlated with 10-year overall survival (OS) rates, in patients with breast cancer. Among patients with breast cancer, those with high HPD expression had worse OS rates than those with low HPD expression. Additionally, when patients were subgrouped by disease stage or grade, those with high HPD expression had worse OS rates than those with low HPD expression for each respective stage or grade.ConclusionsOur findings indicate that HPD may be a useful prognostic predictor, and a potential therapeutic target for patients with breast cancer.     

Expression of Gli1 and Wnt2B correlates with progression and clinical outcome of pancreatic cancer

Objective: The aim of this study was to investigate the expression and clinical significance of Gli1 and Wnt2B in pancreatic cancer. Methods: We have constructed a formalin-fixed paraffin embedded pancreatic tissue microarrays 180cylindrical tissue cores of human pancreatic cancer and its paracancerous nonmalignant pancreatic specimens (NMPs) from 90 patients. Levels of Gli1 and Wnt2B were measured by immunohistochemistry. We analyzed the correlations between the expression of these factors and clinicopathological parameters including prognosis. Results: The expressions of both Gli1 and Wnt2B in human pancreatic cancer tissues were significantly higher than those of normal pancreatic tissues (P=0.000, P=0.004 respectively). The analysis showed that the high cytoplasmic expression levels of Gli1 in pancreatic cancer tissues had significant correlation with lymph node metastasis (P=0.036) and Wnt2B had significant correlation with perineural invasion (P=0.045). Gli1 and Wnt2B have no positive correlation. Survival analysis by Kaplan-Meier demonstrated that elevated Wnt2B expression in cancer tissue predicted worse overall survival (OS) compared with group in lower expression (P=0.024). No correlation was found between the expression of Gli1 and overall survival of pancreatic cancer patients (P>0.05). Conclusions: In conclusion, these results indicate that the high-expression levels of Gli1 and Wnt2B might play a pivotal role during tumorigenesis of pancreatic cancer, and the high expression of Wnt2B might be associated with poor prognosis.