Nonhomologous end joining repair pathway molecules as predictive biomarkers for patients with oral squamous cell carcinoma.

PURPOSE: Nonhomologous end-joining (NHEJ) is critical for the repair of either pathologic double-strand breaks (DSBs) and/or for the repair of physiologic DSBs created during radiotherapy to kill the tumor cell. Therefore, patients with higher expression of NHEJ repair proteins might develop resistance to ionizing radiation, allowing the disease to recur. As cancer of the oral cavity is a serious health problem globally, the present study aimed to examine the expression of Ku70/80, X-ray repair cross-complementing protein 4 (XRCC4) and DNA ligase IV-core molecules of the NHEJ pathway in patients with oral cancer. MATERIALS AND METHODS: Protein expression of Ku70/80, XRCC4, and DNA ligase IV were studied by Immunohistochemistry and mRNA expression of Ku70 and Ku80 were studied using reverse transcription polymerase chain reaction. Data were analyzed statistically using SPSS. RESULTS: A univariate survival analysis revealed an association of Ku70 mRNA with shorter overall survival (OS). While protein expression of XRCC4 showed an association with reduced relapse-free survival and shorter OS. Multivariate survival analysis demonstrated that XRCC4 and DNA ligase IV are independent prognosticators for predicting adverse disease outcomes. CONCLUSION: Strong expression of repair proteins - XRCC4 and DNA ligase IV is associated with unfavorable disease outcome in patients with oral squamous cell carcinoma.

Transcriptome profiling and pathway analysis in squamous cell carcinoma of buccal mucosa.

BACKGROUND: High recurrence and poor overall survival in buccal mucosa squamous cell carcinoma (BMSCC) are not well addressed due to lack of efficient prognostic biomarkers and targeted therapies. To uncover gene candidates for the same, transcriptome profiling has been examined in BMSCC, which is not explored yet. METHODS: We compared 9 BMSCC and 2 normal oral FFPE tissues using Agilent SurePrint G3 Human gene expression v3 microarray chips. The obtained RNA signatures were interrogated in the cancer genome atlas (TCGA) dataset for alteration values and survival data. RESULTS: We found total 237 protein coding RNAs and 85 long non-coding RNAs (lncRNAs) which displayed significant differential expression with criteria of at-least 2 fold change and Benjamini Hochberg FDR < .05. In protein coding RNAs, RUNX3 and EMX2 showed utmost degree of up-regulation and down-regulation, respectively. Likewise, among lncRNAs, ARGFXP2 and lnc-SYCP3-2 displayed highest degree of up-regulation and down-regulation, respectively. Besides, an analysis of the RNA list in TCGA dataset spotted deregulation of 21 genes in both, our cohort and TCGA cohort. Among which, MRTO4 and EIF3J genes, and LINC00310, a lncRNA showed greatest expression alterations. Strikingly, at RNA expression level, up-regulation of two genes, EIF3J and SDCBP, was significantly associated with disease free survival and poor overall survival, respectively. CONCLUSION: Our data documented significant findings to enhance understanding of the disease biology. The proposed RNA candidates (RUNX3, EMX2, MRTO4, EIF3J, SDCBP and LINC00310) may serve as putative therapeutic targets and potential biomarkers for BMSCC diagnosis and prognosis.

Medulloblastoma under Siege: Genetic and Molecular Dissection Concerning Recent Advances in Therapeutic Strategies.

Medulloblastoma (MB) is a devastating illness with unmet therapeutic needs, predominantly cytotoxic and nontargeted approaches. Survivors of MB also suffer from severe treatment-related effects of radiation and cytotoxic chemotherapy keeping mortality rate significant. Recently, four distinct molecular subgroups of MB have been identified (WNT [wingless], SHH [sonic hedgehog], Group 3, and Group 4). Novel subgroup-specific therapies are being explored in the daily treatment of patients as a clinical trial and are an important challenge in the near term for the pediatric neurooncology society. Epigenetic modifiers are also recurrently affected in MB suggesting that epigenetic therapy can be considered in a subset of patients. Moreover, a hint on forefront procedure; tracer of cancer's genetic information entitled "liquid biopsy" in MB is described. This review examines the recent scientific progress in MB research, with a focus on the genes, pathways that drive tumorigenesis and the advances in conventional and targeted therapy. The identification of subgroup-specific, actionable therapeutic targets has the potential to revolutionize therapy for patients with MB and results in significantly enriched overall survival.

Deciphering the potential value of 5-fluorouracil metabolic enzymes in predicting prognosis and treatment response of colorectal cancer patients.

INTRODUCTION: 5-flourouracil (5-FU) is one of the standard chemotherapeutic drugs used today in the treatment of colorectal cancer patients. Disruption of 5-FU metabolic pathway may contribute to altered effectiveness towards 5-FU-based therapy. Hence, the study of 5-FU metabolizing enzymes might have the potential efficacy to predict survival and response to treatment in colorectal cancer patients. MATERIALS AND METHODS: Immunohistochemical localization of 5-FU metabolic enzymes (TS, MTHFR, DPYD, and TP) was evaluated in 143 untreated patients with colorectal cancer; their prognostic and predictive values were also evaluated. RESULTS: Immuno-positivity for TS, MTHFR, DPYD, and TP was observed in 77%, 75%, 88%, and 96% of colorectal cancer patients, respectively. Univariate survival analysis in total patients showed that low DPYD expression significantly predicted adverse overall survival ( P=0.042). Moreover, subgroup of colon cancer patients with low TS expression was associated with unfavorable prognosis. TP expression also emerged as a prognosticator in the subgroup of early and advanced stage patients. Additionally, when effect of co-expression of 5-FU metabolic enzymes was evaluated in total patients, low coexpression of all four proteins was predictive of poor overall survival than for individuals expressing high coexpression of these proteins ( P=0.045). In contrast, none of the 5-FU metabolic enzymes-either singly or on coexpression-emerged as a useful biomarker of potential therapeutic value when evaluated in the subgroup of patients treated with 5-FU alone or 5-FU plus oxaliplatin. CONCLUSION: The above findings suggest that coexpression of 5-FU metabolic enzymes possess significant prognostic value and could be useful biomarkers in colorectal cancer patients.

Significance of expression of suppressor of cytokine signaling proteins: Suppressor of cytokine signaling-1, suppressor of cytokine signaling-2, and suppressor of cytokine signaling-3 in papillary thyroid cancer.

PURPOSE: Uncontrolled cytokine signal transduction largely associated with oncogene activation, can have disastrous biological consequences. The suppressor of cytokine signaling (SOCS) proteins represent one of the mechanisms by which this rampant signaling can be dissipated. Thus, we aimed to study the expression of SOCS-1, SOCS-2, and SOCS-3 in patients having benign thyroid disease and papillary thyroid cancer. MATERIALS AND METHODS: SOCS protein expression was studied in 45 patients with benign thyroid disease and in 83 papillary thyroid cancer patients by immunohistochemistry and their association with clinicopathological characteristics and overall survival in cancer patients were analyzed using SPSS software. RESULTS: Expressions of SOCS proteins were significantly higher in papillary thyroid cancer than in patients having benign disease. SOCS-1 expression was predominantly higher in males (P = 0.004), unilateral tumors (P = 0.030), and noninflammatory conditions (P = 0.028). SOCS-1 expression was also able to predict poor overall survival in subgroup of papillary thyroid cancer patients having larger tumor size (P = 0.013) and advanced stage disease (P = 0.033). Expression of SOCS-2 significantly correlated with tumor size (P = 0.017), extrathyroidal extension (P = 0.000), residual disease (P = 0.043), and treatment (P = 0.007), while preponderance of SOCS-3 expression was observed in males (P = 0.030) and in patients having extrathyroidal extension (P = 0.011) and absence of metastasis (P = 0.032). CONCLUSION: Expression of the studied SOCS proteins may be a consequence of activation of Janus kinase-signal transducers and activators of transcription and other pathways supporting growth and survival of cancer cells that are sustained by several cytokines. Thus, SOCS-1, SOCS-2, and SOCS-3 proteins may directly or indirectly, have important roles in development and pathogenesis of papillary thyroid cancer.

Significance of Interleukin-6 in Papillary Thyroid Carcinoma.

This study sought to reveal the significance of IL-6 in papillary thyroid carcinoma by determining its circulating levels, tumoral protein, and mRNA expressions. As compared to the healthy individuals, serum IL-6 was significantly higher in patients with benign thyroid diseases and PTC. Further, its level was significantly higher in PTC patients as compared to patients with benign thyroid diseases. ROC curves also confirmed a good discriminatory efficacy of serum IL-6 between healthy individuals and patients with benign thyroid diseases and PTC. The circulating IL-6 was significantly associated with poor overall survival in PTC patients. IL-6 immunoreactivity was significantly high in PTC patients as compared to the benign thyroid disease patients. Significantly higher IL-6 mRNA expression was also observed in the primary tumour tissues of PTC patients than the adjacent normal tissues. The protein expression of IL-6 at both the circulating and tissue level correlated with disease aggressiveness in PTC patients. Moreover, a significant positive correlation was observed between the IL-6 protein and mRNA expression in the primary tumours of PTC patients. Finally in conclusion, IL-6 has an important role in thyroid cancer progression. Thus targeting IL-6 signalling can help in clinical management of thyroid carcinoma patients.

Significance of TNF-α and the Adhesion Molecules: L-Selectin and VCAM-1 in Papillary Thyroid Carcinoma.

Circulating levels of TNF-α and the adhesion molecules L-Selectin and VCAM-1 as well as their expression in the primary tumors of patients with benign thyroid diseases and papillary thyroid carcinoma (PTC) have been determined in this study. The serum levels of TNF-α, L-Selectin, and VCAM-1 were significantly higher in patients with both benign thyroid diseases and PTC as compared to the healthy individuals. However, the levels of only TNF-α and L-Selectin, and not VCAM-1, were significantly higher in patients with PTC in comparison to those observed in patients with benign thyroid diseases. Further the expression of TNF-α and L-Selectin was also significantly higher in the primary tumors of PTC patients, relative to the benign thyroid diseases. The expression of L-Selectin and VCAM-1 significantly correlated with aggressive tumor behavior. In PTC patients, the circulating TNF-α levels significantly positively correlated with the levels of L-Selectin, while TNF-α immunoreactivity was significantly associated with VCAM-1 expression. Serum TNF-α was found to be a significant prognosticator for OS in PTC patients. Overall the results signify that the interaction between TNF-α and the adhesion molecules may have a role in thyroid carcinogenesis and understanding this complexity may offer potential therapeutic targets for better management of thyroid cancer.

CD44v6 expression in primary breast carcinoma in western India: a pilot clinicopathologic study.

AIMS AND BACKGROUND: In India, breast cancer is becoming the number one cancer in females. CD44 is believed to play a critical role in the metastatic process, and its spliced forms, especially CD44v6, bestow a metastatic phenotype onto non-metastatic cells. However, the biological significance of CD44v6 in tumor progression remains controversial. Hence, pursuing our interest based on previous observations of a significant association of CD44 standard with advanced stage and poor survival, the present study investigated CD44v6 expression in our series of breast cancer. METHODS AND STUDY DESIGN: For this purpose, 85 untreated primary breast cancer patients were enrolled. CD44v6 was localized immunohistochemically, and its mRNA transcript along with CD44v9 and CD44v10 mRNA were studied by reverse transcriptase polymerase chain reaction. RESULTS: Membranous and/or cytoplasmic staining of CD44v6 was observed in 48% of the primary breast cancers. CD44v6 protein expression showed no significant association with clinical risk factors and survival. At the RNA level, the expression of CD44v6, CD44v9 and CD44v10 in breast cancers was 44%, 22% and 36%, respectively. CD44v6 mRNA expression significantly correlated with CD44v9 (P = 0.013) and CD44v10 (P = 0.0001) but showed no correlation with its protein expression. Furthermore, except for CD44v6 mRNA, none of the other isoforms were associated with clinical risk factors or survival. Loss of CD44v6 mRNA was significantly associated with poor overall survival (P = 0.018). In multivariate overall survival analysis, loss of CD44v6 mRNA expression was a significant independent factor of a poor prognosis (P = 0.045) with a relative risk of 2.10, entering the equation at step three after stage and lymph node status. CONCLUSIONS: Preliminary results suggest an important role of CD44v6 in our series of patients. Down-expression of CD44v6 may be associated with the tumor cell phenotype, facilitating aggressive growth properties that affect the prognosis.

Prognostic significance of molecular markers in oral squamous cell carcinoma: a multivariate analysis.

BACKGROUND: Multiple marker accumulation impacts tumor progression and biologic phenotypes affect clinical outcome of patients with head and neck cancer. Hence, this study investigated a battery of molecular markers that may help to reflect biologic aggressiveness and predict prognosis. METHODS: Epidermal growth factor receptor (EGFR), Stat3, H-ras, c-myc, p53, cyclin D1, p16, Rb, Ki-67, and Bcl-2 were localized immunohistochemically in 135 oral squamous cell carcinoma patients to assess prognostic value. RESULTS: In univariate analysis of total patients, p53, Stat3, and p16 predicted both relapse-free survival (RFS) and overall survival (OS). In Cox multivariate analysis, after adjusting for tumor size, nodal status, and lymphatic permeation, p53 was independently associated with RFS and OS, and p16 with RFS only. In only early-stage patients, in univariate analysis, nuclear Stat3 was significant for RFS and OS. CONCLUSION: Immunostaining of p53, p16, and Stat3 might serve as potential adjuncts in pathologic evaluation of oral tumors to predict risk of relapse.

Expression of C-Myc mRNA in squamous cell carcinoma of the tongue.

BACKGROUND: The aim of this study was to evaluate clinical significance of C-myc mRNA in patients with tongue cancer. METHODS: C-myc mRNA expression was studied by RT-PCR in peripheral blood of 25 tongue cancer patients and 24 controls. C-myc protein expression was studied by immunohistochemistry. RESULTS: In tongue cancer patients, pretherapeutic C-myc mRNA expression was significantly higher as compared to controls. In tumor tissues, a trend of low expression of C-myc mRNA was noted as compared to pretherapeutic blood. The mean pretherapeutic C-myc mRNA level was lower in tobacco-users, in older patients, in keratinizing tumors, in tumors showed lymphocytic infiltration as well as in non-responders as compared to their respective counterparts. C-myc mRNA expression was lower in tumors showed lymphatic permeation and in patients with a habit of tobacco use. Further, low C-myc mRNA expression associated with poor prognosis. C-myc protein expression was noted in 72% of the tumors and an inverse correlation was noted between C-myc protein expression and disease stage. In early stage disease, an inverse correlation was noted while in advanced stage disease, a positive correlation was noted. CONCLUSION: In tongue cancer, downregulation of C-myc mRNA associated with advancement of the disease and worse prognosis.

BRCA1 expression in leukoplakia and carcinoma of the tongue.

BACKGROUND AND OBJECTIVES: Expression of BRCA1 was examined in patients with leukoplakia and carcinoma of the tongue. Its prognostic value was evaluated in patients with tongue cancer. METHODS: Expression of BRCA1 was studied by immunohistochemical localization. Cytoplasmic staining of BRCA1 was observed in both leukoplakia and carcinoma of the tongue. RESULTS: In leukoplakia, 61% and 39% of the patients expressed BRCA1 expression with a staining intensity of 1+ and 2+, respectively. In patients with hyperplasia (67%), BRCA1 expression with a staining intensity of 1+ was 67%; BRCA1 expression with a staining intensity of 2+ was 33%. In patients with dysplasia (33%; mild and moderate), BRCA1 expression with a staining intensity of 1+ and 2+ was 50% each. In carcinoma of the tongue, only 34% of the patients showed BRCA1 expression. In this group, 33% of the tumors exhibited 1+ staining, and only 1% of the tumors expressed 2+ staining. Moreover, BRCA1 expression with a staining intensity of 2+ was significantly higher in patients with dysplasia (50%) than in those with hyperplasia (33%), followed by patients with squamous cell carcinoma of the tongue (1%). The percentage positivity of BRCA1 expression in tongue cancer patients was significantly lower (34%), as compared with patients with leukoplakia (100%; P = 0.000001). A significant positive correlation was noted between BRCA1 and c-myc (P = 0.012). Univariate survival analysis by log-rank test and multivariate survival analysis by Cox regression showed that BRCA1 expression was the most significant prognostic factor predicting relapse-free survival of early-stage patients. CONCLUSIONS: Subcellular localization of the BRCA1 gene product provided evidence of its involvement in the pathogenesis of tongue tumors.

Prognostic significance of biomarkers in squamous cell carcinoma of the tongue: multivariate analysis.

BACKGROUND AND OBJECTIVES: Expression of a panel of biomarkers, such as p53, Bcl-2, Cyclin D1, c-myc, p21ras, c-erb B2, cytokeratin-19 (CK-19), and factor VIII-related antigen (FVIII-RA), was studied together in anterior tongue tumors from the oral cavity and in posterior tongue tumors from the oropharynx of patients with early- and locally advanced-stage disease, to evaluate their prognostic value. METHODS: The expression of the above-mentioned biomarkers was studied by immunohistochemical localization. RESULTS: In this study, 18%, 26%, 62%, 75%, 73%, 50%, and 29% of the tumors exhibited p53, Bcl-2, Cyclin D1, c-myc, p21ras, c-erb B2, and CK-19 expression, respectively. Twenty percent of the tumors had a microvessel count of >0.0. The expression of these biomarkers was also correlated with clinicopathologic parameters. In early-stage patients with a tobacco habit, who showed borderline significance for relapse-free survival by Kaplan-Meier survival analysis, this turned out to be significant, with the general linear model univariate survival analysis. In the total group, disease stage emerged as the most significant prognostic factor, followed by c-myc, when Cox forward stepwise regression and general linear model multivariate survival analysis were performed. However, Cyclin D1, which was significant by Cox forward stepwise regression analysis, lost its significance by general linear model multivariate analysis. In patients with early-stage disease, MVC, which was a significant predictor of disease relapse by Cox forward stepwise regression analysis, lost its significance by general linear model analysis because of small number of patients. In patients with locally advanced tongue cancer, multivariate survival analysis of individual biomarkers by both Cox forward stepwise regression and general linear model analysis indicated c-myc expression to be strongly indicative of poor prognosis. However, multivariate analysis of individual markers along with a combination of markers showed that only by Cox forward stepwise regression analysis did the combined expression of markers c-myc, Cyclin D1, and p21ras emerge as a significant independent prognosticator. CONCLUSIONS: Overall stage emerged as the most significant prognostic indicator of disease outcome. Tobacco habit also affected relapse-free survival in patients with early-stage disease. However, immunostaining of c-myc in the tumors of locally advanced-stage tongue cancer patients might be a potential adjunct to clinical stage in the pathologic evaluation of tongue specimens.