Down-regulation of microRNA-182 and microRNA-183 predicts progression of osteosarcoma.

INTRODUCTION: The aim of this study was to investigate the expression levels of microRNA-182 and microRNA-183 and their association with clinicopathological features in patients with osteosarcoma. MATERIAL AND METHODS: Total RNA was purified from samples and noncancerous bone tissues and then quantitative real-time polymerase chain reaction was applied to evaluate the expression levels of microRNAs, and their relationship with clinicopathological features and survival in osteosarcoma patients. RESULTS: Our findings showed that expression of MiR-182 was clearly lower in osteosarcoma bone tissue (mean ± SD: 2.84 ±.07) compared with noncancerous bone tissues (6.23 ±1.72, p = 0.004). On the other hand, lower expression of MiR-183 was seen in osteosarcoma bone tissue (1.43 ±0.59) when compared with normal tissues (4.36 ±2.47, p = 0.036). Decreased expression of MiR-182 was clearly correlated with advanced clinical stage (p = 0.001), metastasis or recurrence (p = 0.024), and large tumor size (p = 0.032). Decreased expression of MiR-183 was associated with advanced TNM stage (p = 0.004), and metastasis or recurrence (p = 0.002). A multivariate Cox proportional hazards model revealed that low expression of MiR-182 and MiR-183 (p = 0.02; p = 0.016), TNM stage (p = 0.04), and metastasis or recurrence (p = 0.03) were significantly associated with poor survival as independent prognostic factors. CONCLUSIONS: These findings suggest that MiR-182 and MiR-183 may be associated with progression and metastasis of osteosarcoma.

Investigation of serum levels and tissue expression of two genes IGFBP-2 and IGFBP-3 act as potential biomarker for predicting the progression and survival in patients with glioblastoma multiforme.

BACKGROUND: Identification of genetic copy number changes in glial tumors is of importance in the context of improved/refined diagnostic, prognostic procedures and therapeutic decision-making. Blood-derived biomarkers, therefore, would be useful as minimally invasive markers that could support diagnosis and enable monitoring of tumour growth and response to treatment. OBJECTIVE: The aim of this study was to evaluate the clinical significance of IGFBP-2/3 in glioblastoma multiforme (GBM) and their value as predictors of survival. METHODS: We examined the plasma levels of IGFBP-2 and IGFBP-3 using ELISA in patient suffering from GBM and controls groups. Furthermore, immunohistochemistry method was used to evaluate the expression levels of these markers. RESULTS: Preoperative plasma levels of IGFBP-2 and IGFBP-3 were markedly higher in glioblastoma patients (mean±SD: 521.5±164.2ng/ml; 402.4±126ng/ml) when compared with healthy controls (301.28±73.12; 244±89.5ng/ml; p<0.001). Immunohistochemical results indicated that the median H score for glioblastoma tissues was higher when compared with normal tissues. The mean scores for IGFBP-2 expression in glioblastoma was higher than normal tissues (p<0.001). Our result showed that the median H score for glioblastoma tissues was higher when compared with normal tissue for IGFBP-3 expression. The mean scores for glioblastoma tissues was higher than normal tissues (p<0.001). We also evaluated whether plasma IGFBP-2 and IGFBP-3 levels were related to clinical features. The plasma IGFBP-2 level was strongly linked to the patient's age (R=0.769, P=0.001) that were strongly increased in patients with older age (>65), (mean±SD: 594.36±33.3ng/ml). On the other hand, plasma IGFBP-3 level was not correlated with age (P=0.462), sex (P=0.532), and tumor size (P=0.245). Our findings indicated that the tissue IGFBP-2 level was also markedly correlated with the patient's age (R=0.612, P=0.015). On the other hand, tissue IGFBP-3 expression level was not correlated with age (P=0.472), sex (P=0.512), and tumor size (P=0.241). Kaplan-Meier survival and log-rank analysis suggested that patients with high plasma level of IGFBP-2 and tissue expression of IGFBP-2 had shorter overall survival than those with low levels (log-rank test P=0.027; P<0.001). Kaplan-Meier survival and log-rank analysis suggested that patients with high plasma level of IGFBP-3 and tissue expression of IGFBP-3 had shorter overall survival than those with low levels groups (log-rank test P=0.018; P<0.001). CONCLUSION: These data suggest that plasma levels and tissue levels of IGFBP-2 and IGFBP-3 may be as potential biomarkers for predicting the progression and survival in patients with GBM.

Somatic CPEB4 and CPEB1 genes mutations spectrum on the prognostic predictive accuracy in patients with high-grade glioma and their clinical significance.

BACKGROUND: Glioblastoma (grade IV glioma/GBM) is characterized by extremely aggressive invasion and proliferative nature. OBJECTIVE: The main goal of this study was to evaluate the expression patterns of CPEB1 and CPEB4 in glioma patients. METHODS: 41 paraffin-embedded tissue samples with glioma (WHO I-IV) were collected between January 2008 and December 2012 in Tehran, Iran. MRI of patients was done before and within 24 h after surgery and gliomas investigated using quantitative real-time PCR and immunohistochemistry. Kaplan-Meier survival and Cox regression were applied to assess the prognosis of patients. RESULTS: The mRNA level of CPEB4 was strongly increased in tumor tissues (0.67±3.154 vs. 1.671±0.51; P=0.001). Furthermore, CPEB1 mRNA was significantly decreased in tumor tissues compared to normal tissues (2.852±0.587 vs. 1.471±0.862; P=0.025). Our findings showed that CPEB4 levels was markedly increased in patients with advanced grade gliomas (P=0.003). In addition, CPEB1 mRNA levels were not associated with clinicopathological features. Of the 41 cases, high CPEB4 expression was found in 29 patients (70.73%), while 12 cases (29.26%) showed weak expression levels, while the protein expression of CPEB4 were remarkably weak in normal tissues (P=0.001). However, no correlation was found between expression levels of CPEB1 and clinicopathological characteristics. Kaplan-Meier survival and log-rank test indicated that high expression of CPEB4 was correlated with shorter overall survival (log-rank test P<0.001). Furthermore, low expression of CPEB1 was linked to shorter overall survival (log-rank test P=0.021). Multivariate Cox proportional hazards model showed that high CPEB1 (P=0.027), low CPEB4 expressions (P=0.021), and advanced tumor grade (P=0.036) were independent predictor of overall survival. CONCLUSION: Our data indicated expressions levels of CPEB4 and CPEB1 are correlated with overall survival in patients with glioma.

Overexpression of ubiquitin-specific protease 2a (USP2a) and nuclear factor erythroid 2-related factor 2 (Nrf2) in human gliomas.

BACKGROUND: Gliomas are among the most frequent adult primary brain tumors. Recent studies have shown that there are novel opportunities for developing therapeutics by targeting the differentiation and self-renewal features of glioma. OBJECTIVE: The aim of this study was to evaluate the expression levels of USP2a an Nrf2 in patients with glioma and their association with prognosis of gliomas that was detected with immunohistochemical staining. METHODS: In this study, 40 patient's tissue samples with primary gliomas were collected between January 2009 and December 2013. MRI of patients was done before and within 24 h after surgery. USP2a and Nrf2 expression levels were examined by immunohistochemistry. Data were analyzed using the SPSS 16.0, X(2) test, log-rank test and Kaplan-Meier method. RESULTS: Immunohistochemistry indicated that USP2a expression was increased in glioma cells than normal brain tissues. The increased USP2a staining was markedly correlated with advanced tumor grade (P=0.02) and age (P=0.016). Our result showed that Nrf2 expression was significantly higher in glioma cells as compared to normal brain tissues. The high expression level of Nrf2 was markedly linked to age (P=0.007), and tumor grade (P=0.03). Kaplan-Meier survival and log-rank analysis indicated that patients with low expression of USP2a had longer overall survival than those with high levels (log-rank test P<0.001). Moreover, patients with high Nrf2 expression had shorter overall survival than those with low levels (log-rank test P<0.001). In the univariate analysis, the high expression of Nrf2 and USP2a (P=0.004; P=0.006), age (P=0.025), and tumor grade (P=0.001) were correlated with poor survival. Multivariate Cox proportional hazards model indicated that, high Nrf2 and USP2a staining (P=0.001; P=0.003), advanced tumor grade (P=0.01) and age (P=0.033) were independent predictor of overall survival. CONCLUSION: In summary, the result of this study showed USP2a and Nrf2 may be as prognostic marker in patients with gliomas.

Downregulation of miR-185 and upregulation of miR-218 expression may be potential diagnostic and prognostic biomarkers of human chondrosarcoma.

Increasing evidence has confirmed that dysregulation of microRNAs (miRNAs) can contribute to the progression and metastasis of human tumors. Chondrosarcoma is the most common primary malignant bone tumor in adults and has no effective systemic treatment, and patients with this disease have poor survival. Thus, it is important to find new diagnostic markers and improve treatment options. In the current study, we are interested to examine the role of miR-185 and miR-218 expression in patients with chondrosarcoma using real-time PCR. Moreover, the association of the two miRNAs with clinicopathological features and prognosis was evaluated. Survival and Cox proportional hazards analyses were performed to find the association of miR-185 expression and miR-218 levels with prognosis in the patients. Our results indicated that the miR-185 expression was significantly downexpressed in clinical chondrosarcoma bone tissues compared with adjacent normal tissues (P = 0.001). MiR-218 expression level was increased in clinical chondrosarcoma bone tissue than those adjacent normal tissues (P = 0.001). Decreased expression of miR-185 showed remarkable correlation with advanced tumor stage (P = 0.019), tumor grade (P < 0.001), and distant metastasis (P = 0.001). Moreover, high expression of miR-218 was strongly correlated with advanced tumor stage (P = 0.014), tumor grade (P < 0.001), and distant metastasis (P = 0.002). Kaplan-Meier survival analysis revealed that the low miR-185 expression group and the high miR-218 expression group had remarkably shorter overall survival (log-rank test P = 0.007, P = 0.004). The multivariate Cox proportional hazards model indicated that decreased expression of miR-185 and increased expression of miR-218 (P = 0.017, P = 0.012), advanced tumor stage (P = 0.006, P = 0.012), tumor grade (P = 0.032, P = 0.016), and distant metastasis (P = 0.004, P = 0.015) were independently related to overall survival in patients with chondrosarcoma. In conclusion, downregulation of miR-185 and upregulation of miR-218 can be associated with progression of chondrosarcoma and also both of them may act as tumor suppressor genes in chondrosarcoma.

Tissue expression levels of miR-29b and miR-422a in children, adolescents, and young adults' age groups and their association with prediction of poor prognosis in human osteosarcoma.

Osteosarcoma is the most common type of bone cancer in children and adolescents. MicroRNAs (miRNAs) play important roles in the development, differentiation, and function of different cell types and in the pathogenesis of various human diseases. miRNAs are differentially expressed in normal and cancer cells. The investigation of miRNA expression between healthy subjects and patients with osteosarcoma is crucial for future clinical trials. In this study, the expression levels of miRNAs were detected by qRT-PCR. Correlation between expression levels of tow miRNAs and different clinicopathological characteristics were analyzed using the χ (2) test. Survival rate was detected using the log-rank test and Kaplan-Meier method. qRT-PCR was shown that expression levels of miR-29b and miR-422a were strongly decreased in osteosarcoma bone tissue compared with noncancerous bone tissues. Our result indicated that the low expression levels of miR-29b and miR-422a showed strong correlation with large tumor size (P = 0.20; 0.029), advanced TNM stage (P = 0.001; 0.012), distant metastasis (P = 0.008; 0.019), and grade of tumor (P = 0.009; 0.016). Kaplan-Meier survival analysis showed that the low expressions of miR-29b/miR-422a were correlated with shorter time overall survival (log-rank test, P = 0.009; P = 0.013). Moreover, multivariate Cox proportional hazards model indicated that miR-29b and miR-422a (P = 0.024; P = 0.016) were independent prognostic markers of overall survival of patients. Our result indicated that downregulation of miR-29b and miR-422a may be linked to the prediction of poor prognosis, indicating that miR-29b and miR-422a may be a valuable prognostic marker for osteosarcoma patients.

Expression and prognostic value of the aldehyde dehydrogenase 1 (ALDH1) and N-myc downstream regulated gene 2 (NDRG2) as potential markers in human astrocytomas.

In this study, immunohistochemical analysis was used to evaluate the expression of ALDH1 and NDRG2 in astrocytoma tissue samples and normal brain tissues. ALDH1 protein staining displayed that AlDH1 expression was not detectable in eight astrocytoma tissues (8/36) and in all of normal brain tissues. There was a significant difference between ALDH1 expression and WHO grades (P = 0.03). Furthermore, no correlation was determined between expression levels of ALDH1 and other clinicopathological characteristics including age, sex, and tumor size. Immunohistochemistry showed that a high level of NDRG2 protein expression was markedly detected in normal brain tissues and expression of NDRG2 protein was significantly decreased in astrocytoma tissues. There was a significant association between pathological grading and NDRG2 expression level (P < 0.001, Table 1), but no correlation was determined between expression levels of NDRG2 and other clinicopathological characteristics including age, sex, and tumor size. We also obtained detailed follow-up data and evaluated the association of ALDH1/NDRG2 expressions with overall survival. Kaplan-Meier survival and log-rank analysis indicated that the patients with high proportion of ALDH1-positive cells and low proportion of NDRG2-positive had shorter overall survival (P < 0.001; P = 0.001). Univariate analysis indicated that the high proportion of ALDH1-positive cells (P < 0.001), the low proportion of NDRG2-positive cells (P = 0.009), and the advanced grade (P < 0.005) were markedly linked to the prognosis in patients. Furthermore, in the multivariate analysis, ALDH1 cells' expression (P = 0.012), low proportion of NDRG2-positive cells (P = 0.025), and advanced grade (P < 0.03) were linked to poor overall survival. Our results suggest that NDRG2 expression is related to decreased survival rates and NDRG2 may be a potential marker in the astrocytoma prognosis. NDRG2 may be a potential marker in the astrocytoma prognosis. ALDH1 expression was related to advanced pathological grade and survival rate in astrocytoma patients.

Evaluation of gene expression level of CDC5L and MACC1 in poor prognosis and progression of osteosarcoma.

Current evidences have indicated that osteosarcoma is strongly associated with abnormal genetic and epigenetic changes that lead to the abnormal expression of oncogenes or methylation of tumor suppressor genes. In the present study, MACC1 and CDC5L mRNA levels in the patients with osteosarcoma were evaluated using quantitative real-time PCR. Our results demonstrated that CDC5L mRNA levels were higher in tumor tissues than in adjacent normal tissues (2.713 ± 0.738 vs. 1.071 ± 0.629; P < 0.05). Moreover, MACC1 was upregulated in tumor bone tissues than in adjacent normal tissues (3.221 ± 0. 624 vs. 1.427 ± 0.456; P < 0.05). Our result demonstrated that high expression of CDC5L was significantly related to advanced TNM stage (P = 0.032). No significant difference was determined between CDC5L mRNA expression and other clinicopathological parameters including age, gender, tumor diameter, location, tumor grade, and histological type. In addition, overexpression of MACC1 was strongly correlated with advanced TNM stage (P = 0.027) and high tumor grade (P = 0.035). Our findings indicated that mRNA level of CDC5L is correlated with advanced TNM stage, and MACC1 may be involved in progression of osteosarcoma.

Down-regulated MicroRNA 148b expression as predictive biomarker and its prognostic significance associated with clinicopathological features in non-small-cell lung cancer patients.

BACKGROUND: Lung cancer is most common and is the leading cause of cancer-related death in both men and women worldwide. Understanding of the molecular mechanisms underlying non-small cell lung cancer (NSCLC) development and progression are important. In the present study, we investigated the potential role of miR-148b expression analysis as potential lung cancer biomarker with the correlation of circulating miR-148b to clinicopathological features. METHODS: A total of 104 NSCLC patients were diagnosed and cancer tissues together with adjacent normal tissues were evaluated. Quantitative Real-time PCR method was utilized to evaluate the expression levels of miR-148b. In addition, we investigated to clarify the relationship of miR-148b with clinicopathological features and survival in patients with NSCLC. RESULTS: Our findings showed that miR-148b was downregulated in tumor tissues when compared with corresponding adjacent normal lung tissues (0.34 ± 0.13 vs. 1.00 ± 0.57, P < 0.05). Moreover decreased expression of miR-148b was significantly related to TNM stage (P = 0.001) and lymph node-metastasis (P = 0.023). This findings suggested that miR-148b was down-regulated in NSCLC patients and may play a key role as a tumor suppressor gene in NSCLC. Kaplan-Meier survival analysis and log-rank test suggested that low-expression group of patients had significantly shorter overall survival than high-expression group (log-rank test: P = 0.031). Multivariate Cox proportional hazards model analysis indicated that low miR-148b expression was independently linked to poor survival of patients with NSCLC (HR = 3.215, 95 % CI: 1.543-10.621, P = 0.021) and other factors were not significant independent predictor of survival in patients with NSCLC. CONCLUSION: Our findings demonstrated that miR-148b may play a role as independent prognostic factor for patients with NSCLC.