Cancer Biomarkers - Volume 24, issue 2

Authors: Ziai, Hedyeh | Alenazi, Abdulrahman | Hearn, Matthew | O’Connell, Daniel A. | Puttagunta, Lakshmi | Barber, Brittany | Harris, Jeffrey R. | Seikaly, Hadi | Biron, Vincent L.

Article Type: Research Article

Abstract: BACKGROUND: The role of molecular biomarkers in oropharyngeal squamous cell carcinoma (OPSCC) has recently been increasingly recognized. There is conflicting evidence in the literature with regards to the prognostic value of p53 and Bcl-xL. OBJECTIVE: The purpose of this study was to investigate the association between p53 and Bcl-xL expression profiles and survival outcomes in OPSCC. METHODS: Patients diagnosed with OPSCC and treated with curative intent between 1998 and 2009 were included in the study. Patient demographics, disease, treatment, and oncologic outcomes were collected prospectively. A tissue microarray (TMA) from patients’ biopsies or …surgical specimens was retrospectively constructed. The expression levels of p53, Bcl-xL, and p16 were digitally quantified and correlated to patient survival outcomes. RESULTS: One hundred and sixty-six patients were included (mean age 56.7 years; standard deviation (SD) ± 10.0; 78% male). High expression of Bcl-xL (p = 0.04) was significantly associated with nodal disease at presentation, and decreased overall survival (OS) (p = 0.04). Combined expression of low Bcl-xL and low p53 conferred a survival advantage in non-smokers (p = 0.04). Multivariate analysis supported smoking and p16 status as independent prognosticators for OS. CONCLUSIONS: This study suggests that biomarker profiling using Bcl-xL and p53 levels may be of prognostic value in select patients with OPSCC. Show more

Keywords: Oropharyngeal cancer, squamous cell carcinoma, p53, Bcl-XL, biomarkers, survival, recurrence

DOI: 10.3233/CBM-182106

Citation: Cancer Biomarkers, vol. 24, no. 2, pp. 141-151, 2019

Authors: Li, Hang | Pan, Xiang | Gui, Yaoting | Quan, Jing | Li, Zuwei | Zhao, Liwen | Guan, Xin | Xu, Jinling | Xu, Weijie | Lai, Yongqing

Article Type: Research Article

Abstract: OBJECTIVE: Renal cell carcinoma (RCC) is one of the most common genitourinary cancers, and advanced RCC usually leads to poor prognosis. Therefore, identifying novel biomarkers for predicting the progression and prognosis of RCC is essential. The present study aims to evaluate the clinical value of miR-183-5p in RCC development and prognosis after surgery. MATERIALS AND METHODS: We enrolled a total of 284 patients who received partial or radical nephrectomy from April 2003 to May 2013 at a single institution. The clinical and pathological characteristics of the patients were collected, including age, gender, tumor size, tumor stage, …as well as follow-up information. The expression levels of miR-183-5p of all the patients were calculated from FFPE specimens. Cox regression analyses were performed to approve the effect of miR-183-5p expression on patient survival. Kaplan-Meier method was used to analyze the patient survival curves. RESULTS: After controlling for gender, age, tumor size and tumor stage in the multivariate analysis, we found that high expression of miR-183-5p was independently associated lower overall survival (HR = 0.550, 95% CI = 0.364–0.832, p = 0.005). The Kaplan-Meier analysis also showed that patients with high expression of miR-183-5p had a significantly poor prognosis (p = 0.006). These results was verified by analyzing the data of 506 cases from The Cancer Genome Atlas database (TCGA). CONCLUSION: Our results indicated that the high miR-183-5p expression is an independent factor for predicting RCC’s worse prognosis. Show more

Keywords: MicroRNAs, miR-183-5p, renal cell cancer, oncogene, prognosis biomarkers

DOI: 10.3233/CBM-182047

Citation: Cancer Biomarkers, vol. 24, no. 2, pp. 153-158, 2019

Authors: Wang, Jin-Rong | Liu, Bin | Zhou, Lei | Huang, Yue-Xin

Article Type: Research Article

Abstract: BACKGROUND: A growing body of studies have demonstrated the aberrant expression of microRNAs (miRNAs) contributes to human tumor metastasis. MicroRNA-124-3p (miR-124-3p), which is down-regulated in various cancers, has been found to be involved in several signaling pathways relevant to tumor cell migration and invasion. However, the roles of miR-124-3p in human bladder cancer remain unclear. This study aims to investigate the functional significance of miR-124-3p and to understand how it targets the integrin receptor, and thus affects the progression of human bladder cancer. METHODS: Clinical specimens from 36 patients and three human bladder cancer cell lines …were analyzed for miR-124-3p and integrin α 3 (ITGA3) . To investigate the effects of miR-124-3p and ITGA3 on proliferation of bladder cancer cells, the MTT assay, colon-formation assay and flow cytometry were performed. In addition, wound healing assay and transwell assay were carried out to examine the migration and invasion of the bladder cancer cells transfected with miR-124-3p mimics or si-ITGA3. The luciferase reporter assay, quantitative real-time polymerase chain reaction (qRT-PCR) and western blot were applied to validate the miR-124-3p directly binding with ITGA3. Finally, western blot was used to examine the expression level of the proteins involved in FAK/PI3K/AKT and FAK/Src signal pathway as well as epithelial-mesenchymal transition (EMT) process. RESULTS: The down-regulation of miR-124-3p and up-regulation of ITGA3 were observed in clinical specimens and bladder cancer cell lines. Overexpression of miR-124-3p or silencing ITGA3 inhibited tumor cell migration and invasion. Luciferase assay confirmed miR-124-3p directly targets ITGA3, and western blot suggested that miR-124-3p plays a crucial role in the EMT and metastasis of human bladder cancer through FAK/PI3K/AKT and FAK/Src signaling mechanism. Also, by targeting ITGA3, miR-124-3p can modulate the expression of N- and E-cadherin, and thus inhibit the EMT. CONCLUSIONS: By targeting ITGA3 and downstream FAK/PI3K/AKT and FAK/Src signaling pathways, miR-124-3p suppresses cell migration and invasion in bladder cancer. Our study reasonably speculates that miR-124-3p can be potentially developed as a therapeutic target and prognostic biomarker for bladder cancer. Show more

Keywords: Bladder cancer, miR-124-3p, ITGA3, FAK/PI3K/AKT, EMT

DOI: 10.3233/CBM-182000

Citation: Cancer Biomarkers, vol. 24, no. 2, pp. 159-172, 2019

Authors: Zhou, Zhijian | Ma, Jing

Article Type: Research Article

Abstract: MicroRNAs (miRNAs) have been demonstrated that play a critical role in tumorigenesis. The aim of this study is to identify the functional role of miR-378 in cholangiocarcinoma (CCA). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the expression levels of miR-378 in human CCA tissue samples and CCA cell lines. The receiver operating characteristic (ROC) curve was established, and the area under the ROC curve (AUC) was calculated to estimate the capacity of miR-378 in distinguishing CCA patients with different TNM stages. Kaplan-Meier survival analysis and Cox regression assay were performed to explore the prognostic value of miR-378. …Cell proliferation capacity was assessed by MTT assay. Cell migration and invasion were identified by Transwell assays. miR-378 was significantly elevated in CCA tissues when compared with adjacent normal tissues, and in CCA cell lines compared to HIBEC cells. And we found that the expression of miR-378 was significantly associated with TNM stage (P = 0.030) and lymph node metastasis (P = 0.018). ROC curve analysis result showed miR-378 could distinguish CCA patients with TNM stages III and IV from those with stages I and II, with the AUC was 0.816. Patients with high expression of miR-378 had a shorter overall survival rate (Log-rank P = 0.030). The miR-378 was proven to be an independent prognostic predictor for the CCA patients (HR = 1.735, 95% CI = 1.007–2.988, P = 0.041). Downregulation of miR-378 could inhibit cell proliferation, migration, and invasion. These results indicated that miR-378 function as an oncogene and promote CCA cells proliferation, migration, and invasion. The miR-378 could be a novel prognostic marker for CCA. Show more

Keywords: microRNA-378, cholangiocarcinoma, prognosis, proliferation, migration, invasion

DOI: 10.3233/CBM-181980

Citation: Cancer Biomarkers, vol. 24, no. 2, pp. 173-181, 2019

Authors: Shi, Shengjun | Tian, Binqun

Article Type: Research Article

Abstract: BACKGROUND: Bladder cancer is one of the most common genitourinary malignancies, with a high rate of recurrence and progression. The prognosis for patients with bladder cancer, especially muscle-invasive bladder cancer, remains poor despite systemic therapy. OBJECTIVE: To explore the underlying disease mechanisms and identify more effective biomarkers for bladder cancer. METHODS: Weighted gene co-expression network analysis (WGCNA) and protein-protein interaction (PPI) network analysis were applied to identify hub genes correlated with the bladder cancer progression. Survival analyses were then conducted to identify potential biomarkers correlated with the prognosis of bladder cancer. Finally, validation …and analysis of these potential biomarkers were conducted by a series of bioinformatics analyses. RESULTS: Based on the results of weighted gene co-expression network analysis and protein-protein interaction network analysis, ten hub genes closely correlated with bladder cancer progression were identified in the relevant module. Survival analyses of these genes indicated that elevated expressions of six potential biomarkers (COL3A1, FN1, COL5A1, FBN1, COL6A1 and THBS2 ) were significantly associated with a worse overall survival. Furthermore, these 6 potential biomarkers were validated in association with the progression of bladder cancer. Bladder cancer samples with higher expression of these genes were most significantly enriched in gene set associated with ECM-receptor interaction. CONCLUSIONS: This study identified several biomarkers associated with bladder cancer progression and prognosis. As novel findings, these may have important clinical implications for diagnosis, treatment and prognosis prediction. Show more

Keywords: Bladder cancer, weighted gene co-expression network analysis (WGCNA), biomarkers, progression, prognosis

DOI: 10.3233/CBM-181940

Citation: Cancer Biomarkers, vol. 24, no. 2, pp. 183-193, 2019

Authors: Feng, Youfan | Zhang, Yuxia | Wei, Xiaofang | Zhang, Qike

Article Type: Research Article

Abstract: OBJECTIVE: Human multiple myeloma (MM) is a kind of common tumor in middle-aged and elderly people, in which the osteolytic lesion is formed mainly through inhibiting osteoblast (OB) differentiation and promoting osteoclast (OC) differentiation. Dickkopf-1 (DKK1) is a soluble Wnt inhibitor, which has an important correlation with the pathogenesis of human MM. Therefore, the correlations of DKK1 with pathogenesis and prognosis of human MM were investigated in this study. METHODS: The DKK1 expression in tissues and serum of myeloma patients was detected via immunohistochemistry and enzyme-linked immunosorbent assay (ELISA). Correlation between DKK1 expression and survival time …of patients was analyzed via Kaplan-Meier analysis. To further study the mechanism of DKK1 expression in pathogenesis and prognosis of human MM, MM cells were treated with DKK1 neutralizing antibody (BHQ880) or transfected with DKK1-small-interfering ribonucleic acid (siRNA) to study its effects on OB differentiation, osteocalcin level, β -catenin and interleukin-6 (IL-6) secretion. Moreover, the effect of DKK1-siRNA transfection on the activity of U266 cells was detected via methyl thiazolyl tetrazolium (MTT) assay. RESULTS: The DKK1 expression in tissues and serum of myeloma patients was significantly higher than that in control group (p < 0.01). In terms of survival time, the median survival time (45 months) in patients with low DKK1 expression was significantly longer than that in patients with high DKK1 expression (only 22 months). The DKK1 neutralizing antibody (BHQ880) and DKK1-siRNA significantly reduced the DKK1 level in MM cells, promoted the OB differentiation, increased the osteocalcin deposition, promoted the β -catenin expression and decreased the IL-6 expression and β -catenin phosphorylation. DKK1-siRNA could also reduce the proliferative activity of MM cells. CONCLUSION: DKK1 is closely related to the pathogenesis and prognosis of human MM, which might be a potential biomarker for the diagnosis of MM. Show more

Keywords: DKK1, human multiple myeloma, pathogenesis, prognosis

DOI: 10.3233/CBM-181909

Citation: Cancer Biomarkers, vol. 24, no. 2, pp. 195-201, 2019

Authors: Yang, Ying | Zhang, Zongduan | Wu, Zhengzheng | Lin, Wei | Yu, Man

Article Type: Research Article

Abstract: BACKGROUND: Long noncoding RNA MIAT expression is related to the development of some diseases. However, the role of MIAT in melanoma was has seldom been studied. OBJECTIVE: To investigate the effect of the lncRNA MIAT on melanoma cells. METHOD: Microarray was used to analyze the lncRNAs expression in tissue samples. The expression of the lncRNA MIAT was detected by qRT-PCR. A CCK-8 assay was used to assess cell viability, and cell counting was used to analyze cell proliferation. Wound healing and Transwell invasion assays were used to detect the migration and invasion abilities, …respectively, of melanoma cells. Western blotting was performed to explore the molecular mechanisms of MIAT in melanoma. RESULTS: The lncRNA MIAT was overexpressed in melanoma. The overexpression of MIAT promoted cell proliferation, cell invasion and migration, while the knockdown of MIAT expression got the opposite results. MIAT significantly upregulated the phosphorylation of PI3K and AKT and promoted cMyc and cyclin D1 protein expression. CONCLUSION: LncRNA MIAT was a key factor to promote cell invasion, migration and proliferation through the PI3K/AKT signaling pathway. These findings may give us a potential way to treat melanoma. Show more

Keywords: lncRNA, MIAT, melanoma, migration, invasion, PI3K/AKT signaling pathway

DOI: 10.3233/CBM-181869

Citation: Cancer Biomarkers, vol. 24, no. 2, pp. 203-211, 2019

Authors: Liu, Longyang | Zeng, Zhaoyang | Yi, Juanjuan | Zuo, Liu | Lv, Jin | Yuan, Jianhuan | Lin, Zhongqiu | Luo, Rongcheng | Feng, Xin

Article Type: Research Article

Abstract: OBJECTIVES: To investigate TCF4 expression in epithelial ovarian cancer, and to explore its correlation with clinicopathological parameters and clinical prognosis of epithelial ovarian cancer. METHODS: From 2009 to 2017, 188 cases of paraffin-embedded epithelial ovarian cancer tissues and 41 paratumor ovarian tissues which had been confirmed at the memorial hospital of Sun Yat-sen University were collected in this study, and the expression of TCF4 was performed by immunohistochemistry using a polyclonal antibody specific for TCF4. RESULTS: The expression of TCF4 protein was associated with disease progression free survival and overall survival in epithelial …ovarian cancer patients; and TCF4 overexpression was associated with age, FIGO stage, lymph node metastasis, intraperitoneal metastasis, intestinal metastasis, vital status, intraperitoneal recurrence, and serum CA153. Moreover, in a multivariate Cox regression analysis TCF4 overexpression was an indeed independent prognostic factor in epithelial ovarian cancer. CONCLUSIONS: TCF4 may play an oncogenic role in epithelial ovarian cancer, and TCF4 is a useful independent prognostic biomarker of epithelial ovarian cancer, and it may provide a candidate target therapy treatment in future. Show more

Keywords: TCF4, ovarian cancer, expression, clinical significance

DOI: 10.3233/CBM-181849

Citation: Cancer Biomarkers, vol. 24, no. 2, pp. 213-221, 2019

Authors: Wakinoue, Shiro | Chano, Tokuhiro | Amano, Tsukuru | Isono, Takahiro | Kimura, Fuminori | Kushima, Ryoji | Murakami, Takashi

Article Type: Research Article

Abstract: BACKGROUND: Endometrioid ovarian carcinoma and clear cell ovarian carcinoma are both classified as endometriosis-associated ovarian cancer (EAOC). Despite the high rates of recurrence and mortality of EAOC, no prognostic biomarkers have been determined. ADP-ribosylation factor-like protein 4C (ARL4C) has been reported to be involved in various tumor progression processes, but its clinical significance for predicting prognosis in EAOC cases has never been studied. OBJECTIVE: The present study aimed to determine the clinical significance of ARL4C expression in EAOC prognosis. METHODS: ARL4C expression was semi-quantitatively evaluated via immunohistochemistry in 61 EAOC patients, and the …correlations between ARL4C expression and clinicopathological data and survival were statistically analyzed. RESULTS: Thirty-six (59%) cases had high levels of ARL4C, which was related to worse 5-year overall survival (OS) (log-rank test, p = 0.036). In multivariate Cox proportional hazard model, high ARL4C expression was a significantly independent predictive factor for worse 5-year OS (hazard ratio = 12.048, p = 0.0201) and 5-year PFS (hazard ratio = 8.130, p = 0.0036). CONCLUSIONS: ARL4C is a biomarker for worse prognosis and a novel therapeutic target in EAOC. Show more

Keywords: ADP-ribosylation factor-like protein 4C, endometriosis-associated ovarian cancer, clear cell ovarian carcinoma, endometrioid ovarian carcinoma

DOI: 10.3233/CBM-181836

Citation: Cancer Biomarkers, vol. 24, no. 2, pp. 223-229, 2019

Authors: Yan, Lin | Yu, Hai-Hua | Liu, Yuan-Shui | Wang, Yan-Sen | Zhao, Wen-Hua

Article Type: Research Article

Abstract: BACKGROUND: Colorectal cancer (CRC) is the most common malignant disease worldwide and thus new therapeutic approaches are needed. 5-Fluorouracil (5-FU) remains the most widely used agent to treat colorectal cancer (CRC). However, its clinical efficacy is currently limited by the development of drug resistance. Esculetin (EST), a coumarin, was found to have anti-proliferative and anti-migration activity in cancer. OBJECTIVE: This research aims to evaluated the influence and possible mechanism of EST on the proliferation, migration and epithelial-mesenchymal transition of CRC cell lines. MATERIALS AND METHODS: Human CRC cell lines HT-29, SW480, HCT-116, and …Caco-2 were treated with various concentrations of EST (0.2, 2, 20, 200, 2000 μ g/ml) or 5-FU (0.1, 1, 10, 100, 1000 μ g/ml) for 48 h, and cell viability was determined by the MTT and CCK-8 assay. The motility of HCT-116 cells was detected by scratch assay. Western blot was applied to detect the protein expression. Besides, levels of Wnt3a and VEGF in HCT-116 cell culture medium supernatant were analyzed by ELISA. The anti-tumor effect was detected with HCT-116 subcutaneous tumor bearing tumor model by monitoring the tumor vomume in vivo . Finally, the tumoral expression of VEGF was measured by immunohistochemistry, and the expression of Ki67, PCNA, β -catenin, c-Myc, Cyclin D1, MMP2 and MMP7 was measured by Western blot analysis. RESULTS: EST inhibited HCT-116 cell proliferation in a dose-dependent manner. Western blot analysis revealed that EST decreased the expression of Ki67, PCNA, N-cadherin, E-cadherin, vimentin, fibronectin, β -catenin, c-Myc, Cyclin D1, MMP2 and MMP7. Furthermore, EST reduced the release of Wnt3a and VEGF into HCT-116 cells culture medium. After EST treatment, the tumor volume was significant smaller than that of the control group, and the tumoral levels of VEGF were decreased. Moreover, western blot analysis indicated that the expression of Ki67, PCNA, β -catenin, c-Myc, Cyclin D1, MMP2 and MMP7 were also significantly decreased after treated with EST. In addition, in vitro and in vivo anti-tumor results demonstrated that EST combined with 5-FU could increase the inhibitory effect of 5-FU on HCT-116 cells proliferation, migration and epithelial-mesenchymal transition. CONCLUSIONS: EST enhances the inhibitory effect of 5-FU on the proliferation, migration and epithelial-mesenchymal transition of CRC. Show more

Keywords: EST, CRC, proliferation, migration, epithelial-mesenchymal transition

DOI: 10.3233/CBM-181764

Citation: Cancer Biomarkers, vol. 24, no. 2, pp. 231-240, 2019

Authors: Pasha, Heba F. | Mohamed, Randa H. | Radwan, Mohamed I.

Article Type: Research Article

Abstract: BACKGROUND: DNA methylation status is one of the most prevalent molecular alterations in human cancers. Identification of powerful diagnostic and prognostic biomarkers for hepatocellular carcinoma (HCC) without a biopsy is urgently required. OBJECTIVE: The purpose of this study was to determine the methylation status of RASSF1A and SOCS-1genes as a non-invasive biomarker for HCC identification and prognosis. METHODS: Methylation specific-PCR technique was performed to recognize the methylation status of RASSF1A and SOCS-1 genes in 100 patients with HCC, 100 patients with liver cirrhosis (LC) but without HCC were considered as cirrhotic liver control …group and 100 healthy control. RESULTS: Methylation of RASSF1A and SOCS-1 genes were detected in 40% and 38% of HCC patients respectively, 14% and 20% of LC patients respectively. Methylation of SOCS-1 gene in peripheral blood of healthy control was 23%. Methylation of RASSF1A gene was associated with age, tumor size, vascular invasion and α fetoprotein (AFP), while SOCS-1 gene methylation was significantly associated with tumor size and AFP. Furthermore, using RASSF1A/ SOCS-1/ AFP panel improve diagnostic sensitivity for HCC 86% and specificity of 75%. CONCLUSION: RASSF1A and SOCS1 genes methylation status may play an important role in the process of hepatocarcinogenesis and may be used as diagnostic and prognostic noninvasive biomarkers for HCC when combined with serum AFP. Show more

Keywords: Hepatocellular carcinoma, methylation, SOCS-1, RASSF1A

DOI: 10.3233/CBM-181638

Citation: Cancer Biomarkers, vol. 24, no. 2, pp. 241-247, 2019

Authors: Ji, Bing | Huang, Youmin | Gu, Ting | Zhang, Li’e | Li, Guohong | Zhang, Changgeng

Article Type: Research Article

Abstract: BACKGROUND: Plasma carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9), and cancer antigen 72-4 (CA72-4) are common markers which are useful in the diagnosis and prognosis of GC. However, their sensitivity and specificity in GC remain unsatisfactory. Identification of cancer diagnosed-biomarkers would be of great value. OBJECTIVE: Evaluate the diagnostic and prognostic value of LINC00086 and miR-214 in GC. METHODS: In this study, we determined the expression of LINC00086 and miR-214 in GC by qRT-PCR. Additionally, we investigated the relationship between various clinicopathological features of GC patients and LINC00086 or miR-214 expression, and evaluated the diagnostic …and prognostic value of LINC00086 and miR-214 in GC. RESULTS: In this study, we found that plasma LINC00086 expression was significantly lower, whereas plasma miR-214 expression was significantly higher in GC patients than in normal individuals. LINC00086 and miR-214 exhibited high sensitivity and specificity in diagnosing GC. Additionally, GC patients with low LINC00086 or high miR-214 expression were likely to have larger tumors, lymphatic metastasis, larger TNM stage, and higher CEA and CA19-9 levels. Moreover, GC patients with low LINC00086 or high miR-214 expression showed lower survival rates. Lymphatic metastasis, LINC00086, and miR-214 are independent factors affecting patient diagnosis. CONCLUSIONS: LINC00086 and miR-214 are potentially diagnostic and prognostic markers for GC. Show more

Keywords: Gastric cancer, LINC00086, miR-214, diagnosis, prognosis

DOI: 10.3233/CBM-181486

Citation: Cancer Biomarkers, vol. 24, no. 2, pp. 249-255, 2019