Cancer Biomarkers - Volume 20, issue 4

Authors: Yang, Liu | Wang, Tiejun | Zhang, Jun | Wang, Xuxia

Article Type: Research Article

Abstract: BACKGROUND: Epithelial-mesenchymal transition (EMT) is a complicated process that has been implicated in cancer progression and metastasis as well as the formation of many tissues and organs. BTB/POZ domain-containing protein 7 (BTBD7) is reported to regulate transcriptional factors and involved in the process of invasion and metastasis of some malignant tumors. Additionally, our preliminary studies have confirmed that BTBD7 expression was significantly correlated with Slug expression and poor prognosis of primary salivary adenoid cystic carcinoma (SACC). On this basis, this study further investigated function of BTBD7 in the invasion and metastasis of SACC in vitro , which may be …a possible target of gene therapy in the future. METHODS: The expression of BTBD7 and Slug were both examined in SACC-LM and SACC-83 cell lines by immunofluorescence staining. High invasive SACC-LM cells were transfected with BTBD7 siRNA and the expression levels of BTBD7 and Slug were detected in both gene and protein levels by qRT-PCR and western blot analysis. Assays were performed to survey cell migration, invasion and proliferation capabilities with BTBD7 silencing. RESULTS: BTBD7 and Slug proteins were detected in SACC-LM and SACC-83 cell lines. BTBD7 silencing down-regulated the expression of Slug and MMP9 meanwhile up-regulated the expression of E-cadherin in SACC-LM cells, the migration and invasion abilities of cells were obviously suppressed but with no influence on cell proliferation. CONCLUSIONS: BTBD7 silencing inhibited EMT through regulation of Slug expression in SACC-LM cells and might act as a potential molecular target for gene therapy of SACC. Show more

Keywords: BTBD7, Slug, salivary adenoid cystic carcinoma, RNA interference

DOI: 10.3233/CBM-170262

Citation: Cancer Biomarkers, vol. 20, no. 4, pp. 461-468, 2017

Authors: Zhao, Ping | Meng, Meng | Xu, Bin | Dong, Aiping | Ni, Guangzhen | Lu, Lianfang

Article Type: Research Article

Abstract: MUC1, a membrane tethered mucin glycoprotein, is overexpressed in > 60% of human pancreatic cancers (PCs), and is associated with poor prognosis and enhanced metastasis. Here, we report the effect of silencing MUC1 expression on the growth, migration and invasive ability of pancreatic cancer cells, and explored its mechanisms. We observed that siRNA mediated suppression of the MUC1 expression significantly reduced invasive and migrative capability and induced apoptosis of the pancreatic cancer PANC-1 cells. We found that Slug was inhibited in the MUC1 siRNA transfected PANC-1 cells (MUC1 siRNA/PANC-1 cells). Expression of PUMA and E-cadherin was increased …in the MUC1 siRNA/PANC-1 cells. PANC-1 cells overexpressing full long Slug gene (when transfected with Slug cDNA plasmid) significantly inhibited PUMA and E-cadherin expression in the MUC1 siRNA/PANC-1 cells. Silencing PUMA expression inhibited apoptosis in the MUC1 siRNA transfected PANC-1 cells (MUC1 siRNA/PANC-1 cells). Silencing E-cadherin expression restored the invasion and migration ability in the MUC1 siRNA/PANC-1 cells. We therefore concluded that silencing MUC1 expression inhibited migration and invasion, and induced apoptosis of PANC-1 cells via downregulation of Slug and upregulation of Slug dependent PUMA and E-cadherin expression. MUC1 could serve as a potential therapeutic target in pancreatic cancer. Show more

Keywords: Pancreatic cancer, invasion, MUC1, Slug

DOI: 10.3233/CBM-170297

Citation: Cancer Biomarkers, vol. 20, no. 4, pp. 469-476, 2017

Authors: Kim, Soo-Jin | Kim, Eunhee | Rim, Kyung-Taek

Article Type: Research Article

Abstract: Testing carcinogenicity caused by chemicals requires a noninvasive tool that can be used before autopsy because autopsy takes a long time. We investigated whether non-small cell lung cancer related gene mutations could be detected in cell-free DNA in plasma by Insight Onco TM next-generation sequencing, which is a fast and sensitive method. Adenoma formation was confirmed in urethane-injected 17-week-old mice. Seven single nucleotide polymorphisms, such as Cdkn2a and Vegfa, were selected. Mutant-enriched Insight Onco TM NGS and normal NGS were performed on genomic DNA. The results demonstrate that …Insight Onco TM NGS detected Cdkn2a and Vegfa SNPs at 0.05%. The sensitivity of Insight Onco TM NGS was twice higher than that of normal NGS. In this analysis, the Cdkn2a gene mutation was detected not only in two genomic DNA samples of lung tissue from the 11th week of urethane injection but also in two cell-free DNA samples. In addition, the Vegfa gene mutation was detected not only in three genomic DNA samples of lung tissue of injection but also in one cell-free DNA sample, showing 33% concordance. Our results confirm that Insight Onco TM NGS is a rapid and sensitive detection method that enables lung cancer-associated gene mutations to be detected in cell-free DNA before the end of the carcinogenicity test. Show more

Keywords: Cell-free DNA, lung tumor, non-invasive, plasma, quantification

DOI: 10.3233/CBM-170303

Citation: Cancer Biomarkers, vol. 20, no. 4, pp. 477-485, 2017

Authors: Chen, Jing | Cao, Shun-Wang | Cai, Zhen | Zheng, Lei | Wang, Qian

Article Type: Research Article

Abstract: BACKGROUND: Epithelial-mesenchymal transition (EMT) plays a crucial role in circulating tumor cells (CTCs) dissemination and cancer metastasis. OBJECTIVE: To investigate the EMT phenotypes of CTCs in hepatocellular carcinoma (HCC) patients and the clinical utility in the early diagnosis of HCC metastasis and progression. METHODS: We retrospectively analyzed the count and EMT classification of CTCs detected by the CanPatrol ® platform in 195 HCC patients. The clinical relevance with other pathological features was statistically evaluated. RESULTS: CTCs were detected in 95% of the 195 HCC patients with …a range of 0–86 CTCs. Total CTCs numbers were correlated with BCLC stages, metastasis and serum AFP levels. The AUC of the ROC curve was 0.861 (95% CI: 0.782–0.940) in discriminating metastatic HCC patients with non-metastatic patients. Epithelial, hybrid and mesenchymal CTCs were found in about 53%, 83% and 57% patients, respectively. The proportion of hybrid and mesenchymal CTCs was associated with ages, BCLC stages, metastasis and AFP levels. Besides, recurrent HCC patients presented higher CTCs count and increased hybrid and mesenchymal CTCs. CONCLUSIONS: CTCs count and EMT classification are correlated with clinical stages and metastasis of HCC, suggesting that they may be potential markers for the early diagnosis of HCC metastasis and progression. Show more

Keywords: Circulating tumor cells, hepatocellular carcinoma, epithelial-mesenchymal transition, cancer metastasis

DOI: 10.3233/CBM-170315

Citation: Cancer Biomarkers, vol. 20, no. 4, pp. 487-498, 2017

Authors: Jiang, Lei | Wang, Wen-Jun | Li, Zhan-Wu | Wang, Xiao-Zhou

Article Type: Research Article

Abstract: BACKGROUND: Gastric cancer is one of the most common malignancies worldwide. Recent studies reported that Piwil3 was overexpressed in various cancers, including gastric cancer (GC). This study was intended to investigate its function and mechanism in GC progress. METHODS: Quantitative real time PCR(RT-PCR) and western blotting assays were utilized to measure mRNA and protein expression levels, respectively. SiRNA transfection was performed to suppress the expression of Piwil3. CCK-8 assay, cell invasion and migration assays were used to determine the cell proliferative, cell invasive and migratory ability. RESULTS: The expression of Piwil3 was significantly …increased in GC tissues compared with matched normal tissues. The specific siRNA significantly inhibited the protein and mRNA expressions of Piwil3, and effectively inhibited the proliferation and induced G0/G1 phase arrest in GC cells. Downregulation of Piwil3 significantly suppressed the migration and invasion of GC cells. Moreover, the downregulation of Piwil3 also significantly suppressed the tumor volumes in nude mice. Mechanism investigation showed that the downregulation of Piwil3 significantly decreased the mRNA and protein expressions of metastasis-related genes, including RhoC, MTA1, MMP2 and MMP9, and also modulated the phosphorylation levels of JAK2 and STAT3 but not their protein levels. CONCLUSIONS: These findings indicate that overexpression of Piwil3 promotes the proliferation, migration and invasion of GC cells partially through JAK2/STAT3 signal pathway. Show more

Keywords: Piwil3, gastric cancer, proliferation, metastasis, JAK2/STAT3

DOI: 10.3233/CBM-170324

Citation: Cancer Biomarkers, vol. 20, no. 4, pp. 499-509, 2017

Authors: Gou, Xun | Zhao, Xiyan | Wang, Zhengrong

Article Type: Research Article

Abstract: BACKGROUND: Long noncoding RNAs (lncRNA) have been verified to be involved in hepatocellular carcinoma (HCC) progression. However, the potential biologic function of PVT1 in HCC is not still fully known. METHODS: PVT1 and miR-214 were detected by qRT-PCR assays in HCC tissues and adjacent normal tissues. CCK8, cell colony and transwell invasion assays were performed to evaluate cell proliferation and invasion abilities. Western-blot assay was performed to detect the protein of E-cadherin and Vimentin. QRT-PCR, RNA immunoprecipitation (RIP) and chromatin immunoprecipitation (ChIP) assays demonstrated PVT1 regulated miR-214 expression. RESULTS: The results showed that …PVT1 was increased in HCC tissues and higher PVT1 expression was associated with tumor size, histological differentiation grade and advanced TNM stage. Furthermore, we revealed that PVT1 promoted cell proliferation and invasion in HCC. RIP and ChIP assays demonstrated that PVT1 significantly inhibited miR-214 expression by interacting with enhancer of zeste homolog 2 (EZH2). CONCLUSIONS: Thus, these results demonstrated that PVT1/EZH2/miR-214 regulatory pathway might serve as new target for HCC treatment. Show more

Keywords: Hepatocellular carcinoma, PVT1, enhancer of zeste homolog 2, miR-214, long non-coding RNA

DOI: 10.3233/CBM-170331

Citation: Cancer Biomarkers, vol. 20, no. 4, pp. 511-519, 2017

Authors: Chen, Jian | Yang, Li | Wang, Xiongwei

Article Type: Research Article

Abstract: BACKGROUND: MicroRNAs (miRNAs) have been demonstrated to play an important role in the development and progression of various types of cancer including glioblastoma (GBM). OBJECTIVE: The aim of this study was to investigate the expression pattern and prognostic significance of serum miR-203 in patients with GBM. METHODS: miR-203 extracted from cell culture medium and serum samples was detected by real-time PCR. The correlation between serum miR-203 expression as well as clinicopathological characteristics and patient survival was determined. RESULTS: The expression level of miR-203 was remarkably reduced in the GBM cells …and their culture medium. Serum miR-203 expression was significantly decreased in GBM patients compared with low grade glioma (LGG) patients and healthy controls. In addition, serum miR-203 discriminated GBM patients from LGG patients and healthy subjects. Chi-squared analysis showed that a significant correlation was found between low serum miR-203 expression and larger tumor size as well as lower Karnofsky Performance Scale scores. Patients with lower serum miR-203 suffered poorer overall survival (OS) and progression free survival (PFS). Multivariate analysis indicated that low miR-203 expression is an independent prognostic factor for poor OS in GBM patients. CONCLUSIONS: These data demonstrate that serum miR-203 expression might serve as a potential prognostic indicator of GBM. Show more

Keywords: Glioblastoma, miR-203, prognosis, real-time PCR

DOI: 10.3233/CBM-170335

Citation: Cancer Biomarkers, vol. 20, no. 4, pp. 521-526, 2017

Authors: Mei, Li-Li | Qiu, Yun-Tan | Huang, Meng-Bing | Wang, Wen-Jun | Bai, Jie | Shi, Zhi-Zhou

Article Type: Research Article

Abstract: miR-99a is down-regulated in esophageal squamous cell carcinoma (ESCC), however the role and underlying mechanism are still unknown. We aim to explore the role and mechanism of miR-99a down-regulation in ESCC. The expression of miR-99a in ESCC tissues and cell lines was detected by Human miRNA Microarrays and Real-time PCR. The effects of miR-99a on cell proliferation, migration and invasion were determined by Cell Counting Kit-8 (CCK-8) assay, transwell migration and invasion assay. Target gene of miR-99a were analyzed by target prediction software and validated by Real-time PCR and Western blotting assay. Our microarray results and four Gene Expression Omnibus …(GEO) datasets showed lower expression level of miR-99a in ESCC tissues. Overexpression of miR-99a using mimics significantly suppressed cell proliferation, and decreased expressions of CCND1, CCNA2 and CCNE1. We also found that enhanced miR-99a significantly inhibited migration, invasion and epithelial-mesenchymal transition (EMT) of ESCC cells, and down-regulated EMT associated transcription factor Slug, and MMPs including MMP2, MMP7 and MMP13. TargetScan predicted insulin-like growth factor 1 receptor (IGF1R) as the cadidate target gene of miR-99a, and western blotting confirmed the negative correlation between miR-99a and IGF1R. Importantly, we further found that knockdown of IGF1R also significantly inhibited the proliferation, migration, invasion and slug-induced EMT of ESCC cells, and reduced the cell cycle regulatory proteins and MMPs. In conclusion, our findings suggested that loss of miR-99a in ESCC promoted the tumor cell proliferation, migration, invasion and slug-induced EMT through activating IGF1R signaling pathway. Show more

Keywords: miR-99a, proliferation, epithelial-mesenchymal transition, insulin-like growth factor 1 receptor, esophageal squamous cell carcinoma

DOI: 10.3233/CBM-170345

Citation: Cancer Biomarkers, vol. 20, no. 4, pp. 527-537, 2017

Authors: Shi, Qin | Zhou, Zhan | Ye, Naishu | Chen, Qiaolin | Zheng, Xiuxia | Fang, Minshan

Article Type: Research Article

Abstract: BACKGROUND: MicroRNAs (miRNAs) emerge as important regulators involved in malignant progression in some tumors. MiR-181a has been found to function as a tumor suppressor in some tumors including non-small cell lung cancer (NSCLC). However, the functional role of miR-181a in NSCLC still needed to be investigated. METHODS: The expression of miR-181a were determined by qRT-PCR, the association between miR-181a and clinicopathological data were performed by chi-square test and survival analysis were evaluated by Kaplan-Meier curve and log rank test. Cell proliferation and invasion were assessed by CCK8, cell colony formation and transwell assays. Luciferase reporter …assay demonstrated that CDK1 was a target of miR-181a. Western blot assay detected the relative protein expression. RESULTS: In the study, our results showed that miR-181a was significantly down-regulated in non-small cell lung cancer (NSCLC) tissues and cell lines. MiR-181 expression levels were significantly associated with histological grade, N status and TNM stage in the patients and lower miR-181a predicted a poor prognosis in NSCLC patients. Furthermore, upregulation of miR-181a significantly suppressed the NSCLC cell proliferation, colony formation, and cell invasion capacities. Moreover, upregulation of miR-181a inhibited CyclinB1 and CyclinD1 expression in NSCLC cells. Luciferase activity assay results demonstrated CDK1 was a direct target of miR-181a and miR-181a inhibited cell proliferation by regulating the mRNA and protein levels of CDK1 in NSCLC cells. CONCLUSION: These data suggested that miR-181a plays a tumor suppressor and may be a potential therapeutic target for NSCLC patients. Show more

Keywords: Non-small cell lung cancer, miR-181a, CDK1, cell proliferation

DOI: 10.3233/CBM-170350

Citation: Cancer Biomarkers, vol. 20, no. 4, pp. 539-546, 2017

Authors: Liu, Suoning | Suo, Jian | Wang, Chunxi | Sun, Xuan | Wang, Daguang | He, Liang | Zhang, Yang | Li, Wei

Article Type: Research Article

Abstract: BACKGROUND: An overwhelming amount of evidence has emerged suggesting that dysregulated microRNAs (miRNAs) play crucial roles in tumorigenesis. OBJECTIVE: The study was to analyze tissue/serum miR-144 expression in gastric cancer and then evaluate their potential to predict the prognosis of gastric cancer. METHODS: We examined miR-144 levels in tissues and peripheral blood samples from 96 gastric cancer patients using real-time PCR. Then the association between tissue/serum miR-144 levels and clinicopathological parameters was determined. RESULTS: The expression levels of miR-144 were significantly down-regulated in the cancerous tissue and serum samples from …gastric cancer patients. Serum miR-144 was able to differentiate the gastric cancer patients from healthy controls with high accuracy. In addition, tissue and serum miR-144 levels were both associated with clinical stage and lymph node metastasis. Moreover, patients with lower tissue or serum miR-144 suffered worse 5 year overall survival and disease free survival. CONCLUSIONS: Taken together, our data support the potential clinical value of tissue and serum miR-144 as prognostic biomarkers in gastric cancer. Show more

Keywords: Biomarker, miR-144, prognosis, gastric cancer

DOI: 10.3233/CBM-170351

Citation: Cancer Biomarkers, vol. 20, no. 4, pp. 547-552, 2017