Cancer Biomarkers

Clinical significance of microRNA (miR)-15a in human esophageal squamous cell carcinoma (ESCC) remains unclear.

To evaluate the expression level of miR-15a and to determine its potential for diagnosis and prognosis in ESCC.

Quantitative reverse transcription polymerase chain reaction was performed to examine the expression levels of miR-15a in ESCC tissues and patients' sera. The diagnostic and prognostic implications of serum miR-15a level in human ESCC were further evaluated.

Expression levels of miR-15a in ESCC tissues and patients' sera were significantly decreased (both P< 0.001). Additionally, serum miR-15a had an optimal diagnostic cut-off point (2.29) for ESCC with sensitivity of 86.36% and specificity of 100.00%. Moreover, low serum miR-15a level more frequently occurred in ESCC patients with advanced tumor-node-metastasis, T and N stages (all P= 0.01) and poor tumor differentiation (P= 0.03). The Kaplan-Meier curve showed that low miR-15a expression was significantly associated with shorter overall survival (OS) and disease-free survival (DFS) of ESCC patients (both P< 0.001). Further multivariate analysis identified miR-15a as an independent prognostic factor for both OS and DFS (both P= 0.01).

Decreased expression of miR-15a may play a crucial role in ESCC development and progression. Serum miR-15a level could be used as a potential diagnostic and prognostic marker in clinics.

Although the red cell distribution width (RDW) has been reported as a reliable predictor of prognosis in several types of cancer, to our knowledge the prognostic value of RDW in gastric carcinoma has not been studied, so far.

We aimed to investigate the role of red cell distribution width (RDW) in predicting prognosis in gastric cancer patients.

All gastric cancer patients who underwent curative surgery (n= 172, 110M/62F) over a five-year study period were evaluated. Data on demographics, preoperative RDW levels, tumor characteristics (early stage: I and II, advanced stage: IIIA-B-C), disease-free (DFS) and overall survival (OS) were retrospectively reviewed. Patients were classified as high RDW group (RDW ≥ 16, n= 62) or low RDW group (RDW < 16, n= 110).

Overall mortality and postoperative 60-day mortality in both groups were 55% and 14%, respectively. A borderline significant association between RDW (0.063) and mortality was noted. Preoperative RDW levels were significantly higher in patients with short-term mortality(17.9 ± 4.3 vs. 16 ± 3.2, p= 0.015). In high RDW group, the incidence of advanced gastric cancer was significantly higher (75 vs. 51%, p= 0.002), whereas DFS (0.035) and OS (p= 0.04) were lower.

The frequency of advanced cancer was high in patients with high RDWvalues. High RDW values were strongly associated with short-term mortality although only a borderline relationship with overall survival was observed.

Cholangiocarcinoma (CCA), a malignant tumor of the biliary epithelium, is a tumor with an ineffective diagnosis and poor prognosis. We have previously identified an overexpression of orosomucoid 2(ORM2) along with CCA development in hamsters using a proteomics technique.

To evaluate plasma ORM2 as a candidate risk biomarker for CCA in humans.

Overexpression of ORM2 in CCA patients was assessed by western blotting and immunohistochemistry. The diagnostic efficacy of ORM2 was investigated in the plasma of patients with hepatobiliary diseases - including 46 cholangitis patients and 70 CCA patients - compared with 20 healthy individuals, using enzyme-linked immunosorbent assay (ELISA).

Overexpression of ORM2 was observed in the cytoplasm of bile duct tumor cells and in the adjacent normal hepatocytes at a much higher intensity than in normal bile duct cells. Western blot analysis revealed that its expression was significantly higher in the plasma of CCA patients compared with that of healthy individuals (P < 0.01). ELISA showed that plasma ORM2 levels were significantly elevated in CCA and cholangitis groups compared with healthy individuals (P < 0.0001). The sensitivity and specificity of ORM2 in distinguishing CCA patients from healthy patients were 92.86% and 73.68%, respectively.

Plasma ORM2 could serve as a new risk marker for CCA.

The miR-212 was among the top differentially expressed miRNAs in pancreatic ductal adenocarcinoma (PDAC).

The aim of this study was to investigate the expression of miR-212 in PDAC and evaluate its correlation with major clinicopathologic features and patients' survival.

Fluorescence in situ hybridization (FISH) was adopted to examine miRNA expression in 45 pancreatic cancer and 20 normal pancreatic tissues. The relationship of miR-212 expression with clinicopathologic parameters and clinical outcome was evaluated.

miR-212 was confirmed to have significantly higher expression in PDAC compared with normal pancreatic tissues (51.1% vs 10%, p< 0.01). High expression of miR-212 was significantly associated with tumor size (p = 0.048) and tumor stage (p = 0.023). Moreover, in univariant analysis, patients with high expression of miR-212 demonstrate significantly poorer overall survival (p= 0.02).

High expression of miR-212 in PDAC is associated with shorter overall survival. It may be not only a potential prognostic marker, but also a possible therapeutic target in PDAC.

Neutrophil gelatinase-associated lipocalin (NGAL) has been proved as a sensitive biomarker in acute and chronic renal injury. Renal impairment is a common complication of multiple myeloma (MM). We attempt to assess the value of NGAL for the early and accurate diagnosis of renal injury in MM patients.

Forty-five MM patients with CKD stage I to V(MM-renal group), 20 MM patients with normal kidney function (MM-non-renal group), and 37 healthy volunteers (healthy control) were compared for serum and urinary NGAL and other renal injury biomarkers (Creatinine[CRE]; Cystatin-C [CysC]; N-acetyl-beta-D-glucosaminidase [NAG]). Other biomarkers reflect the inflammation and tumor burden like high-sensitivity C-reactive protein(hs-CRP) and urine free light chain were also detected.

Among the biomarkers of renal injury, the assessment of serum CysC, CRE and NGAL was a reliable tool to distinguish MM-renal from MM-non-renal. Both serum and urinary NGAL levels were higher in MM-renal patients than in MM-non-renal or healthy controls (187.10 (45.60-699.60) vs 136.70(47.70-216.50) vs 117.7 (69.3-192.3), P< 0.01; 37.50 (6.30-412.10) vs 18.00(0.50-66.50) vs 11.2 (0.9-69.1), P< 0.01). Univariate analysis showed that both serum (Odds Ratio = 1.009; 95%CI 1.002-1.017; P= 0.018) and urinary NGAL (Odds Ratio = 1.038; 95%CI 1.003-1.073; P= 0.031) as well as serum CysC (Odds Ratio = 9.875; 95%CI 1.685-57.882; P= 0.011) were strong predictors for the risk of renal injury in MM patients. Moreover, the urinary NGAL level was negatively correlated with estimated glomerular filtration rate (eGFR) (r= -0.586, P= 0.00003) and has a tendency towards positive correlation with urine free light chain (r = 0.235, P = 0.124) and hs-CRP (r = 0.379, P = 0.074).

The present study demonstrated that urinary NGAL was not superior to serum NGAL in distinguishing MM-renal group from MM-non-renal group. And it could be considered an independent predictor of renal injury from multiple myeloma reflecting active kidney damage, tumor burden, and inflammation.

Promoter hypermethylation can be a useful biomarker for early detection and prognosis of bladder cancer, monitoring response to treatment and complement classical diagnostic procedures.

The molecular test was performed on DNA from bladder cancer cells in voided urine samples, tumor tissue DNA and normal control DNAs. We aimed to assess the diagnostic potential of epigenetic changes in urine DNA from bladder cancer cases at various clinico-pathological stages of the disease.

The methylation status of 5 genes (p14ARF, p16INK4A, RASSF1A, DAPK, APC) in 113 tumor samples paired with voided urine specimens was analyzed by MSP. We compared the results of methylation analysis with UroVysion test.

The methylation profile in tumor/urine DNA was significantly correlated (p ≤ 0,05) with tumor grade in p14ARF, RASSF1a, APC/p14ARF, APC genes, respectively and with stage in p14ARF, RASSF1a/p14ARF genes, respectively. The results of UroVysion test were in correlation with hypermethylation both in tumor and urine DNA in p14ARF, RASSF1a and APC genes (p = 0,008; 0,02 and 0,04, respectively).

Promoter hypermethylation of tumor suppressor genes is a frequent mechanism in bladder cancer. We found promoter hypermethylation in all grades and stages of all cases examined. Methylation profile of selected suppressor genes may be a potential useful biomarker and enhance early detection of bladder cancer using a noninvasive urine test.

Ovarian cancer is one of the important challenges in the field of gynecologic oncology because of some problems in understanding its etiology and pathogenesis. Receptor for advanced glycation end products(RAGE) is a multiligand trans-membranous receptor which is upregulated in some human cancers. Mechanisms of RAGE involvement in carcinogenesis of ovarian cancer are unknown.

This study aimed to investigate the expression of RAGE in ovarian cancers and its association with clinicopathological characteristics.

The RAGE expression level in ovarian cancer and corresponding noncancerous tissues were analyzed by real time quantitative RT-PCR and immunohistochemistry techniques.

Results indicated that RAGE gene was overexpressed in ovarian cancer tissue compared with adjacent noncancerous tissue (p < 0.001). A significant association between RAGE expression and tumor size (p= 0.04), depth of stromal invasion (p = 0.031), lymphovascular invasion (p = 0.041) and stage of cancer (p = 0.041) was observed. The receiver operating characteristic (ROC) analyses yielded the area under the curve (AUC) values of 0.86 for RAGE in discriminating ovarian cancer samples from non-cancer controls.

In conclusion overexpression of RAGE in ovarian cancer may be a useful biomarker to predict tumor progression.

OCT4 is a key regulator of self-renewal and pluripotency in embryonic stem cells which can potentially encode three spliced variants designated OCT4A, OCT4B and OCT4B1. Based on cancer stem cell concept, it is suggested that the stemness factors misexpressed in cancer cells and potentially is involved in tumorigenesis.

Accordingly, in this study, we investigated the potential expression of OCT4 variants in breast cancer tissues.

A total of 94 tumoral and peritumoral breast specimens were evaluated with respect to the expression of OCT4 variants using quantitative RT-PCR and immunohistochemical (IHC) analysis.

We detected the expression of OCT4 variants in breast tumor tissues with no or very low levels of expression in peritumoral samples of the same patients. While OCT4B was highly expressed in lobular type of breast cancer, OCT4A and OCTB1 variants are highly expressed in low grade (I and II) ductal tumors. Furthermore, the results of this study revealed a considerable association between the expression level of OCT4 variants and the expression of ER, PR, Her2 and P53 factors.

All data demonstrated a distinctive expression pattern of OCT4 spliced variants in different types of breast cancer and provide further evidence for the involvement of embryonic genes in carcinogenesis.

microRNAs (miRNAs) plays an important role in tumor development and progression and act as oncogenes or tumor suppressor genes in the carcinogenesis process. miRNA is stable in serum, and recent studies have demonstrated the feasibility of using circulating miRNA as biomarkers in cancer patients. However, currently, no serum biomarkers for the early diagnosis and prognosis of renal cell carcinoma (RCC) have been reported. Therefore, a new molecular marker for early diagnosis and evaluation of recurrence after surgery is required. Our purpose was to identify miRNA signatures that could distinguish the serum of RCC patients from matched healthy controls and validate identified miRNAs as potential biomarkers for RCC.

Serum samples from 30 RCC patients were collected before and 1 month after surgery. 30 cancer-free blood donor volunteers with no history of any cancer were recruited from the same institute. miR-21 and miR-106a expression levels were determined by real-time PCR.

The serum miR-21 level was significantly higher in RCC patients (median, 8.34) than in healthy control individuals (median, 0.70; p= 0.001). A month after surgery, serum miR-21 levels (median, 0.69) were significantly reduced (p= 0.032). The serum miR-106a level was higher in RCC patients (median, 8.99) compared with controls (median, 0.96; p= 0.000), while miR-106a levels (median, 1.01) were reduced a month after surgery (p= 0.028). The expression level of miR-21 and miR-106 a in RCC patients increased significantly, while miR-21 and miR-106a decreased after surgery. This outcome suggests that serum miR-21 and miR-106a expression level was closely related with kidney cancer tissue.

We conclude that serum miR-21 and miR106a are expected to be molecular markers for RCC.

The sensitivity and specificity of biomarkers which have been used in clinical practice for diagnosis of papillary thyroid carcinoma (PTC) are low, it is essential to develop novel diagnostic and prognostic biomarkers for PTC.

To explore the expressions of miR-940, miR-15a, miR-16 and IL-23 in PTC tissues and plasma and their associations with the clinical characteristics of PTC.

We investigated the expressions of miR-940, miR-15a, miR-16 and IL-23 in plasma and thyroid tissues of PTC, nodular goiter and healthy people with qRT-PCR, and further analyzed the associations between their levels and the clinical characteristics of PTC.

Level of IL-23 expression was higher while levels of miR-940, miR-15a and miR-16 expression in the PTC tissues were lower compared with the nodular goiter tissues and perineoplastic thyroid tissues. And the levels of miR-940, miR-15a, miR-16 and IL-23 expression in the PTC tissues were associated with some clinical characteristics of PTC, including bilateral tumor, multicentricity, extrallyroidal invasion, cervical lymph node metastasis, distant metastasis and clinical advanced stages (III/IV).

Expressions of miR-940, miR-15a, miR-16 and IL-23 in PTC tissues might be useful biomarkers and promising targets in the diagnosis of papillary thyroid carcinoma.