Cancer Biomarkers - Volume 25, issue 1

Authors: Hu, Xiao-Hui | Dai, Jian | Shang, Hou-Lai | Zhao, Ze-Xue | Hao, Yue-Dong

Article Type: Research Article

Abstract: BACKGROUND: Despite the major advances in the treatment, the overall survival of osteosarcoma remains poor. MicroRNAs (miRNAs) are involved in tumorigenesis and progression though modulating their target genes. In the present study, the roles of miR-1285-3p in osteosarcoma was investigated. METHODS: Microarray profiling was applied to distinguish the up and down regulated microRNAs in osteosarcoma. Quantitative real-time PCR (qRT-PCR) assay was performed to detect the expression of miR-1285-3p and YAP1 expression. MTT and transwell assays were carried out to determine the cells proliferation and invasion respectively. Moreover, dual luciferase reporter assay was performed to evaluate the …binding efficiency between miR-1285-3p and the 3’UTR of YAP1. RESULTS: MiR-1285-3p was down regulated in osteosarcoma tissues and cell lines and the reduction of miR-1285-3p expression predicted a poor overall survival of osteosarcoma patients. Ectopic expression of miR-1285-3p inhibited osteosarcoma cell proliferation, colony formation and invasion. In addition, YAP1 was further demonstrated as a direct target of miR-1285-3p. Moreover, overexpression of YAP1 reversed the inhibitory effects of miR-1285-3p on osteosarcoma cells proliferation and invasion. CONCLUSIONS: MiR-1285-3p which was low expressed in osteosarcoma inhibited the proliferation and invasion of osteosarcoma cells via direct targeting YAP1. These results suggested that miR-1285-3p might be a potential therapeutic targets and biomarker in osteosarcoma. Show more

Keywords: miR-1285-3p, YAP1, osteosarcoma, proliferation, invasion, prognosis

DOI: 10.3233/CBM-180013

Citation: Cancer Biomarkers, vol. 25, no. 1, pp. 1-10, 2019

Authors: Zou, Guicheng | Wang, Rui | Wang, Minghui

Article Type: Research Article

Abstract: BACKGROUND: Colorectal cancer (CRC) is a common type of cancer around the world. Detection of microRNA (miRNA) aberration in blood samples is a novel approach for CRC screening. OBJECTIVE: The purpose of this study was to explore the serum miR-497 expression pattern in CRC and examine its potential usefulness as a biomarker for CRC diagnosis and prognosis. METHODS: Serum miR-497 expression was evaluated by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in 110 patients with CRC, 35 cases with colorectal adenoma, 54 cases with colorectal polyps, and 70 healthy individuals. RESULTS: …The expression level of miR-497 was significantly downregulated in CRC tissues compared to the normal tissues based on the data from three independent studies GSE68204, GSE41655 and GSE35834. Compared to healthy controls, the serum miR-497 level was significantly decreased in patient with CRC or benign lesion (colorectal adenoma and polyps). Serum miR-497 level was dramatically increased in the post-operative blood samples from early stage CRC patients. Receiver-operating characteristic (ROC) analysis showed that serum miR-497 had a high sensitivity and specificity for discriminating CRC or precancerous colorectal lesion from normal controls. Moreover, low serum miR-497 expression was closely correlated with aggressive clinical features and shorter overall survival (OS). Kaplan-Meier analyses also revealed that OS was strongly associated with lymph node invasion, TNM stage and histological grade. Furthermore, univariate and multivariate analysis showed serum miR-497 was an independent prognostic factor for CRC. CONCLUSIONS: Collectively, serum miR-497 may serve as a promising biomarker for diagnosis and prognosis of CRC. Show more

Keywords: Colorectal cancer, biomarker, serum miR-497, prognosis

DOI: 10.3233/CBM-181902

Citation: Cancer Biomarkers, vol. 25, no. 1, pp. 11-18, 2019

Authors: Li, Xuelu | Song, Chen | Wang, Kainan | Li, Ning | Sun, Siwen | Li, Na | Zhao, Zuowei | Li, Man

Article Type: Research Article

Abstract: BACKGROUD: Triple-negative breast cancer (TNBC) is associated with an aggressive phenotype and poor prognosis, and the lack of druggable markers leads to the unavailability of targeted therapies. Thus, there is an urgent need to identify potential targets for triple-negative breast cancer. OBJECTIVE: In this study, we aimed to explore the expression of LAPTM4B and p27 kip1 in triple-negative breast cancer, and its clinical significance. METHODS: We analyzed the expression and association of LAPTM4B and p27 kip1 in human breast cancer databases. To analyze the role …of LAPTM4B in the aggressiveness of the human triple-negative breast cancer, the expressions of LAPTM4B were knocked down in MDA-MB-231 and HCC1187 cell lines. Then, cell proliferation, migration and apoptosis were assessed in vitro . Furthermore, the immunohistochemistry examinations of LAPTM4B and p27 kip1 expression were performed using surgical specimens from 188 primary triple-negative breast cancer patients. RESULTS: Through analyses of several independent breast cancer cohorts, we found the correlation of the LAPTM4B and p27 kip1 expression. Remarkably, the knockdown of LAPTM4B restored p27 kip1 expression and inhibited the aggressiveness of breast cancer cells. Meanwhile, the knockdown of p27 kip1 relieved the suppression of cell migration. Consistent with the analyses of human breast cancer cohorts, the immunohistochemistry results showed that the expression levels of LAPTM4B and p27 kip1 were correlated in 188 triple-negative breast cancer samples (p = 0.019). We also validated that the higher LAPTM4B expression, the lower p27 kip1 expression (p = 0.0001), and the LAPTM4B+ /p27 kip1 - subgroup (p < 0.0001) were poor prognostic indicators, as well as the higher histologic grade (p = 0.0001). In the multivariate Cox regression, p27 kip1 expression was considered as an independent predictor of survival (p < 0.001). CONCLUSIONS: The overexpression of LAPTM4B and the loss of p27 kip1 expression are correlated. Meanwhile, the up-regulated expression of LAPTM4B together with the down-regulated expression of p27 kip1 could classified a group of breast cancer patients with poor prognosis, consequently considered as a potentially prognostic marker and candidate target for therapeutic intervention of triple-negative breast cancer. Show more

Keywords: Triple negative, breast cancer, LAPTM4B, p27kip1, prognosis

DOI: 10.3233/CBM-182094

Citation: Cancer Biomarkers, vol. 25, no. 1, pp. 19-27, 2019

Authors: Patil, Shankargouda | Babu, Niraj | Subbannayya, Tejaswini | Mohan, Sonali V. | Sathe, Gajanan | Solanki, Hitendra S. | Rajagopalan, Pavithra | Patel, Krishna | Advani, Jayshree | Bhandi, Shilpa | Sidransky, David | Chatterjee, Aditi | Gowda, Harsha | Ferrari, Marco

Article Type: Research Article

Abstract: BACKGROUND: Shisha smoking has been associated with multiple diseases including oral cancer. However, a mechanistic study to investigate alteration of secreted proteins in oral cells due to shisha smoking is lacking. OBJECTIVES: Elucidation of differentially secreted proteins by immortalized human normal oral keratinocytes (OKF6/TERT1) upon chronic exposure to shisha. METHODS: OKF6/TERT1 was chronically treated with 0.5% shisha extract for 8 months. Conditioned media from shisha treated (OKF6/TERT1-Shisha) and untreated (OKF6/TERT1-Parental) cells were subjected to TMT-based quantitative proteomic analysis. Bioinformatics analysis of differentially secreted proteins was carried out using SignalP, SecretomeP and TMHMM. Immunoblot validation …of selected proteins was carried out to confirm the proteomics results. RESULTS: Proteomic analysis of OKF6/TERT1-Parental and OKF6/TERT1-Shisha secretome resulted in the identification of 1,598 proteins, of which 218 proteins were found to be differentially secreted (⩾ 1.5-fold; p -value ⩽ 0.05) in shisha treated cells. Bioinformatics analysis using prediction tools showed secretory potential of differentially secreted proteins identified in OKF6/TERT1-Shisha. Western blotting validated the expression of AKR1C2, HSPH1 and MMP9 in OKF6/TERT1-Shisha secretome in agreement with proteomic data. CONCLUSION: This study serves as a useful resource to understand the effect of chronic shisha smoking on the milieu of secreted proteins of oral cells. In vivo studies are warranted to supplement our in vitro data to elucidate the role of these proteins as early diagnostic biomarkers for oral carcinogenesis among shisha smokers. Show more

Keywords: OKF6/TERT1, secretory biomarkers, waterpipe, TMT, oral cancer, proteomics

DOI: 10.3233/CBM-182099

Citation: Cancer Biomarkers, vol. 25, no. 1, pp. 29-41, 2019

Authors: Šestáková, Šárka | Krejčík, Zdeněk | Folta, Adam | Cerovská, Ela | Šálek, Cyril | Merkerová, Michaela Dostálová | Pecherková, Pavla | Ráčil, Zdeněk | Mayer, Jiří | Cetkovský, Petr | Remešová, Hana

Article Type: Research Article

Abstract: BACKGROUND: Aberrant epigenetic patterns are a hallmark of acute myeloid leukemia (AML). Mutations in profound epigenetic regulators DNMT3A and IDH1/2 often occur concurrently in AML. OBJECTIVES: The aim was to analyze DNA methylation, hydroxymethylation and mRNA expression profiles in AML with mutations in DNMT3A and IDH1/2 (individually and in combinations). METHODS: Infinium MethylationEPIC BeadChip (Illumina) covering 850,000 CpGs was utilized. The validation of hydroxy-/methylation data was done by pyrosequencing. HumanHT-12 v4 Expression BeadChip (Illumina) was used for expression examination. RESULTS: Hierarchical clustering analysis of DNA hydroxy-/methylation …data revealed clusters corresponding to DNMT3A and IDH1/2 mutations and CD34+ healthy controls. Samples with concurrent presence of DNMT3A and IDH1/2 mutations displayed mixed DNA hydroxy-/methylation profile with preferential clustering to healthy controls. Numbers and levels of DNA hydroxymethylation were low. Uniformly hypermethylated loci in AML patients with IDH1/2 mutations were enriched for immune response and apoptosis related genes, among which hypermethylation of granzyme B (GZMB ) was found to be associated with inferior overall survival of AML patients (P = 0.035). CONCLUSIONS: Distinct molecular background results in specific DNA hydroxy-/methylation profiles in AML. Site-specific DNA hydroxymethylation changes are much less frequent in AML pathogenesis compared to DNA methylation. Methylation levels of enhancer located upstream GZMB gene might contribute to AML prognostication models. Show more

Keywords: AML, DNMT3A and IDH1/2 mutations, methylation, hydroxymethylation, GZMB, CHFR

DOI: 10.3233/CBM-182176

Citation: Cancer Biomarkers, vol. 25, no. 1, pp. 43-51, 2019

Authors: Botezatu, Anca | Iancu, Iulia V. | Plesa, Adriana | Manda, Dana | Popa, Oana | Bostan, Marinela | Mihaila, Mirela | Albulescu, Adrian | Fudulu, Alina | Vladoiu, Susana V. | Huica, Irina | Dobrescu, Ruxandra | Anton, Gabriela | Badiu, Corin

Article Type: Research Article

Abstract: BACKGROUND : Thyroid carcinoma is the most common endocrine malignancy worldwide. Changes in DNA methylation can cause silencing of normally active genes, especially tumour suppressor genes (TSG) or activation of normally silent genes. OBJECTIVE : The aim of this study is to evaluate the degree of promoter methylation for a panel of markers for thyroid neoplasms and to establish their relationship with thyroid oncogenesis. METHODS : To generate a comprehensive DNA methylation signature of TSGs involved in thyroid neoplasia, we use Human TSG EpiTect Methyl II Signature PCR Array-Qiagen for 24 samples (follicular adenomas and papillary …thyroid carcinomas) compared with normal thyroid tissue. We extended the evaluation for three TSGs (TP73, WIF1, PDLIM4) using qMS-PCR. Statistical analysis was performed with GraphPad Prism. RESULTS : We noted four important genes NEUROG1, ESR1, RUNX3, MLH1, which presented methylated promoter in tumour samples compared to normal. We found new characteristic of thyroid tumours: methylation of TP73, WIF1 and PDLIM4 TSGs, which can contribute to thyroid neoplasia. A significant correlation between BRAF V600E mutation and RET/PTC rearrangements with TIMP3 and CDH13, RARB methylation, respectively was observed. CONCLUSIONS : TSGs promoter hypermethylation is a hallmark of cancer and a test that uses methylation quantification method is suitable for diagnosis and prognosis of thyroid cancer. Show more

Keywords: Thyroid cancer, tumour suppressor genes, promoter methylation, epigenetics

DOI: 10.3233/CBM-182265

Citation: Cancer Biomarkers, vol. 25, no. 1, pp. 53-65, 2019

Authors: Ma, Nan | Zhu, Lizhe | Yang, Liu | Cui, Yuxin | Zhan, Yingzhuan

Article Type: Research Article

Abstract: S100 family is made up of at least 20 calcium-binding proteins which are involved in many cellular processes. The prognostic values of individual S100 member in ovarian cancer patients are still unknown. In this study, we performed a detailed prognostic values of S100 in ovarian cancer. The mRNA expression levels of S100 family in various cancers were analyzed via the Oncomine, and the protein-protein interaction network of S100 family was analyzed via String. The prognostic values of individual S100 member were evaluated via Kaplan-Meier Plotter. The S100 family genes expression and mutation were analyzed via cBioProtal. We observed that the …mRNA expression of most S100 family were overexpressed in ovarian cancer compared with normal tissues. In survival analysis in Kaplan-Meier Plotter, 10 members of S100 family showed significant correlation with overall survival in ovarian cancer patients. The trends of high expression of individual S100 members were nearly the same in different subtype and pathological grades. However, the S100 family genes expression and mutation showed no significant prognostic values in overall survival and disease free survival in ovarian cancer patients. Although, the results need more verification both in clinical trials and fundamental experiments, our study provide new insights for the prognostic function of S100 family in ovarian cancer and might promote development of S100 targeted inhibitors for the new treatment of ovarian cancer. Show more

Keywords: S100, ovarian cancer, mRNA, prognosis, KM-plotter

DOI: 10.3233/CBM-182276

Citation: Cancer Biomarkers, vol. 25, no. 1, pp. 67-78, 2019

Authors: Li, Wei | Wang, Junmei | Zhang, Dongqing | Zhang, Xiting | Xu, Jumei | Zhao, Li

Article Type: Research Article

Abstract: BACKGROUND: Recently, the incidence and mortality of retinoblastoma (RB) have gradually increased. Many studies support the pivotal role of microRNAs (miRNAs) in the pathogenesis of RB. Alternation of microRNA-98 (miR-98) expression has been detected in several cancers, excluding RB. This study was designed to assess the regulatory mechanisms of miR-98 in human RB. METHODS: RT-qPCR and Western blot analysis were used to detect miR-98 and HMGA2 expression. The effects of miR-98 were explored using the CCK-8 and Transwell assays. Dual-luciferase reporter assay was performed to confirm the relationship between miR-98 and HMGA2. RESULTS: …In RB, downregulation of miR-98 was identified. Upregulation of miR-98 inhibited proliferation, invasion and migration of RB cells. Further, HMGA2 was confirmed as a direct target gene of miR-98. And knockdown of HMGA2 suppressed the progression of RB. Moreover, upregulation of HMGA2 reversed the suppressive effects in the development of RB. In addition, miR-98 also showed suppressive effect on EMT and Wnt/β -catenin pathway. CONCLUSION: MiR-98 targets HMGA2 to act as a tumor suppressor in RB by mediating Wnt/β -catenin pathway. Show more

Keywords: miR-98, retinoblastoma, HMGA2, wnt/β-catenin pathway

DOI: 10.3233/CBM-182315

Citation: Cancer Biomarkers, vol. 25, no. 1, pp. 79-88, 2019

Authors: Sun, Xianghong | Cui, Shichao | Fu, Xiaofeng | Liu, Chuan | Wang, Zhi | Liu, Yuanwei

Article Type: Research Article

Abstract: This article has been retracted, and the online PDF has been watermarked ``RETRACTION''. The retraction notice is available at http://doi.org/10.3233/CBM-229009.

Keywords: miR-146-5p, lung cancer, cisplatin, claudin-12, wnt/β-catenin, PI3K/AKT/MAPK signaling pathways

DOI: 10.3233/CBM-182374

Citation: Cancer Biomarkers, vol. 25, no. 1, pp. 89-99, 2019

Authors: Cao, Wenfeng | Zhao, Youguang | Wang, Liang | Huang, Xiaoke

Article Type: Research Article

Abstract: OBJECTIVE: This study investigates expressions of circ0001429, miR-205-5p and vascular endothelial growth factor (VEGFA) in bladder cancer tissues and their effects on the proliferation, migration and apoptosis. METHODS: Arraystar Human CircRNA chip was applied to analyzing the differential expression of circRNA in four bladder cancer tissues and paired adjacent normal bladder tissues. Real time quantitative PCR was utilized to detect the expression of circ0001429 in tissue specimens. Bioinformatics, RNA immunoprecipitation and luciferase reporter assays were used to verify the relationship among circ0001429, miR-205-5p and VEGFA in bladder cancer. Cell propagation was determined by CCK8 assay and …roles of circ0001429 and miR-205-5p in cell migration were verified with transwell migration assay. Flow cytometry and TUNEL staining were conducted to observe the impact on cell apoptosis ability. Xenograft experiment was also performed to validate the influence of circ0001429 on tumor growth in vivo . RESULTS: Expressions of circ0001429 and VEGFA were up-regulated, whereas miR-205-5p expression was down-regulated in bladder cancer tissues in comparison with paired adjacent normal bladder tissues. Circ0001429 enhanced the propagation and metastasis abilities of T24 cells and 5637 cells in vitro , but reduced cell apoptosis. In vivo experiments revealed the inhibitor role of sh-circ0001429 in tumor growth and lung metastasis. Circ0001429 sponged miR-205-5p that targeted VEGFA , thereby modulating the protein level of VEGFA. Meanwhile, miR-205-5p restrained the cell viability and mobility and promoted the apoptosis in bladder cancer. Circ0001429 could accelerate cell propagation, migration and invasiveness through increasing VEGFA expression via miR-205-5p. CONCLUSION: Circ0001429 and VEGFA were highly expressed in bladder cancer, while miR-205-5p were lowly expressed in bladder cancer. The circ0001429 could target at miR-205-5p to regulate VEGFA and promote the development of bladder cancer. Show more

Keywords: Bladder cancer, circ0001429, miR-205-5p, VEGFA

DOI: 10.3233/CBM-182380

Citation: Cancer Biomarkers, vol. 25, no. 1, pp. 101-113, 2019

Authors: Lu, Chang | Jia, Shengnan | Zhao, Shutao | Shao, Xue

Article Type: Research Article

Abstract: BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignancies, and its global morbidity and mortality are increasing. Previous studies confirmed that miR-342 was involved in the development and progression of malignant tumors. However, the relationship between miR-342 and Wnt/β -catenin signaling pathway in HCC remains unknown. MATERIALS AND METHODS: Cell viability was detected by MTT assay. Immunofluorescence staining was used to detect Brdu-positive cells and Western blot was used to detect the apoptotic proteins. Furthermore, linear correlation analysis was used to investigate the possible relationship between miR-342 and the downstream genes of Wnt/β …-catenin signaling pathway in the progression of HCC. RESULTS: Over-expression of miR-342 significantly reduced cell proliferation and obviously increased apoptosis in HCC, while silencing of miR-342 showed an opposite effect on HCC cell proliferation and apoptosis. In addition, we found that the CXCL12 was the target gene of miR-342. This study also demonstrated that miR-342 up-regulation suppressed Wnt/β -catenin signaling pathway by inhibiting CXCL12 expression. CONCLUSION: Up-regulation of miR-342 inhibited cell proliferation and induced cell apoptosis in HCC by inhibiting Wnt/β -catenin signaling pathway, suggesting that miR-342 might act as a promising tumor gene therapeutic target for HCC patients. Show more

Keywords: miR-342, proliferation, apoptosis, CXCL12, wnt/β-catenin

DOI: 10.3233/CBM-192399

Citation: Cancer Biomarkers, vol. 25, no. 1, pp. 115-126, 2019