Cancer Biomarkers - Volume 32, issue 2

Authors: Lai, Peng-Sheng | Chang, Wei-Min | Chen, Ying-Yin | Lin, Yi-Feng | Liao, Hui-Fen | Chen, Chung-Yu

Article Type: Research Article

Abstract: Colorectal cancer (CRC) has become the third most common cause of cancer-related deaths. CRC occurs because of abnormal growth of cells that can invade other tissues and cause distant metastases. Researchers have suggested that aberrant microRNA (miRNA) expression is involved in the initiation and progression of cancers. However, the key miRNAs that regulate the growth and metastasis of CRC remain unclear. The circulating miRNAs from BALB/c mice with CRC CT26 cell implantation were assayed by microarray. Then, Mus musculus (house mouse) mmu-miR-762 mimic and inhibitor were transfected to CT26 cells for analysis of cell viability, invasion, and epithelial-mesenchymal transition (EMT), …cell cycle, and regulatory molecule expression. Human subjects were included for comparison the circulating Homo sapiens (human) has-miR-762 levels in CRC patients and control donors, as well as the patients with and without distant metastasis. The result for miRNA levels in mice with CRC cell implantation indicated that plasma mmu-miR-762 was upregulated. Transfection of mmu-miR-762 mimic to CT26 cells increased cell viability, invasion, and EMT, whereas transfection of mmu-miR-762 inhibitor decreased the above abilities. Cells treated with high-concentration mmu-miR-762 inhibitor induced cell cycle arrest at G0/G1 phase. However, mmu-miR-762 did not cause apoptosis of cells. Western blot analysis showed that mmu-miR-762 mimic transfection upregulated the expression of Wnt-1 and β -catenin, as well as increased the nuclear translocation of β -catenin. Further analysis was performed to demonstrate the correlation of miR-762 with CRC, and blood samples were collected from CRC patients and control donors. The results showed that serum has-miR-762 levels in CRC patients were higher than in control donors. Among the CRC patients (n = 20), six patients with distant metastasis showed higher serum has-miR-762 levels than patients without distant metastasis. Conclusions, the present study suggests that circulating miR-762 might be a potential biomarker for upregulation of CRC cell growth and invasion, and may be accompanied by the Wnt/β -catenin signaling. Show more

Keywords: Colorectal cancer, distant metastasis, epithelial-mesenchymal transition (EMT), microRNA, Wnt/β-catenin

DOI: 10.3233/CBM-203002

Citation: Cancer Biomarkers, vol. 32, no. 2, pp. 111-122, 2021

Authors: Arrieta, Oscar | Hernandez-Martinez, Juan-Manuel | Montes-Servín, Edgar | Heredia, David | Cardona, Andrés F. | Molina-Romero, Camilo | Lara-Mejía, Luis | Diaz-Garcia, Diego | Bahena-Gonzalez, Antonio | Mendoza-Oliva, Dolores L.

Article Type: Research Article

Abstract: BACKGROUND: Few trials have evaluated the utility of liquid biopsies to detect epidermal growth factor receptor mutations (EGFR m) at the time of response evaluation and its association with the clinical characteristics and outcomes of non-small-cell lung cancer (NSCLC) patients. OBJECTIVE: This study aimed to evaluate, in a real-world clinical setting, the prevalence of plasma EGFR m and its association with the clinical characteristics, response and survival outcomes of NSCLC patients under treatment with EGFR-tyrosine kinase inhibitors (EGFR-TKIs). METHODS: This observational study enrolled advanced or metastatic NSCLC patients, with confirmed tumor EGFR m, receiving …treatment with first- or second-generation EGFR-TKIs. Blood samples for the detection of plasma EGFR m were collected at the time of response evaluation and processed using the Target Selector™ assay. The main outcomes were the detection rate of plasma EGFR m, median Progression-Free Survival (PFS) and Overall Survival (OS) according to plasma EGFR mutational status. RESULTS: Of 84 patients, 50 (59.5%) had an EGFR m detected in plasma. After a median follow-up of 21.1 months, 63 patients (75%) had disease progression. The detection rate of plasma EGFR m was significantly higher in patients with disease progression than in patients with partial response or stable disease (68.3% versus 33.3%; P < 0.01). PFS and OS were significantly longer in patients without plasma EGFR m than among patients with plasma EGFR m (14.3 months [95% CI, 9.25–19.39] vs 11.0 months [95% CI, 8.61–13.46]; P = 0.034) and (67.8 months [95% CI, 39.80–95.94] vs 32.0 months [95% CI, 17.12–46.93]; P = 0.006), respectively. A positive finding in LB was associated with the presence of ⩾ 3 more metastatic sites (P = 0.028), elevated serum carcinoembryonic (CEA) at disease progression (P = 0.015), and an increase in CEA with respect to baseline levels (P = 0.038). CONCLUSIONS: In NSCLC patients receiving EGFR-TKIs, the detection of plasma EGFR m at the time of tumor response evaluation is associated with poor clinical outcomes. Show more

Keywords: ctDNA, EGFR-TKI resistance, lung adenocarcinoma, clinical progression, liquid biopsy

DOI: 10.3233/CBM-203164

Citation: Cancer Biomarkers, vol. 32, no. 2, pp. 123-135, 2021

Authors: Guo, Yong | Li, Chunxue | Zhang, Rongrong | Zhan, Yating | Yu, Jinglu | Tu, Jinfu | Zheng, Jianjian

Article Type: Research Article

Abstract: BACKGROUND: Long non-coding RNA-growth arrest specific transcript 5 (lncRNA-GAS5) plays a suppressive role in activated hepatic stellate cells (HSCs). LncRNAs could circulate in the blood in a cell-free form and serve as promising biomarkers for various human diseases. Herein, we investigated the feasibility of using serum GAS5 as a biomarker for liver fibrosis in chronic hepatitis B (CHB) patients and whether promoter methylation was responsible for GAS5 down-regulation. METHODS: Serum GAS5 levels were quantified using quantitative real-time PCR in CHB patients and healthy controls. GAS5 promoter methylation was examined in LX-2 cells and cirrhotic tissues. …RESULTS: Compared with the sera from healthy controls, lower GAS5 levels were found in the sera from CHB patients. Receiver operating characteristic curve analysis indicated that serum GAS5 had a significant diagnostic value for liver fibrosis in CHB patients. Serum GAS5 negatively correlated with HAI scores as well as ALT values in CHB patients. GAS5 was additionally reduced in cirrhotic tissues, associated with its hypermethylation promoter. In LX-2 cells, transforming growth factor-β 1 treatment led to a reduction in GAS5 expression and an increase in promoter methylation. Hypermethylation of GAS5 was blocked down by DNA methyltransferase (DNMT) inhibitor and restored GAS5 inhibited HSC activation including proliferation and collagen production. Further studies confirmed that GAS5 methylation was mediated by DNMT1. CONCLUSION: We demonstrate that epigenetically-regulated serum GAS5 could serve as a potential biomarker in CHB patients. Loss of GAS5 is associated with DNMT1-mediated promoter methylation. Show more

Keywords: GAS5, serum, liver fibrosis, chronic hepatitis B, DNA methylation

DOI: 10.3233/CBM-203169

Citation: Cancer Biomarkers, vol. 32, no. 2, pp. 137-146, 2021

Authors: Silva, Jenilson da | Nogueira, Leudivan | Coelho, Ronald | Deus, Amanda | Khayat, André | Marchi, Rafael | Oliveira, Edivaldo de | Santos, Ana Paula dos | Cavalli, Luciane | Pereira, Silma

Article Type: Research Article

Abstract: BACKGROUND: Penile cancer (PeCa) is a rare disease, but its incidence has increased worldwide, mostly in HPV + patients. Nevertheless, there is still no targeted treatment for this carcinoma. OBJECTIVE: To predict the main signaling pathways involved in penile tumorigenesis and its potential drug targets. METHODS: Genome-wide copy number profiling was performed in 28 PeCa. Integration analysis of CNAs and miRNAs and mRNA targets was performed by DIANA-TarBase v.8. The potential impact of the miRNAs/target genes on biological pathways was assessed by DIANA-miRPath v.3.0. For each miRNA, KEGG pathways …were generated based on the tarbase and microT-CDS algorithms. Pharmaco-miR was used to identify associations between miRNAs and their target genes to predict druggable targets. RESULTS: 269 miRNAs and 2,395 genes were mapped in cytobands with CNAs. The comparison of the miRNAs mapped at these cytobands and the miRNAs that were predicted to regulate the genes also mapped in these regions, resulted in a set of common 35 miRNAs and 292 genes. Enrichment pathway revealed their involvement in five top signaling pathways. EGFR and COX2 were identified as potential druggable targets. CONCLUSION: Our data indicate the potential use of EGFR and COX2 inhibitors as a target treatment for PeCa patients. Show more

Keywords: Genomic profiling, molecular target, biomarkers, human papillomavirus, penile tumor

DOI: 10.3233/CBM-210035

Citation: Cancer Biomarkers, vol. 32, no. 2, pp. 147-160, 2021

Authors: Pieper, Willi | Ignatov, Atanas | Kalinski, Thomas | Haybaeck, Johannes | Czapiewski, Piotr | Nass, Norbert

Article Type: Research Article

Abstract: BACKGROUND: Neuronatin (NNAT) determined by immunohistochemistry is a negative prognostic biomarker for breast cancer, independent of the major clinicopathological markers. OBJECTIVE: Here, we investigated whether NNAT is also a predictive biomarker for pathological remission after neoadjuvant chemotherapy. METHODS : One hundred and four breast cancer patients, treated with systemic neoadjuvant chemotherapy were included in this retrospective study. NNAT was detected in formaldehyde fixed, paraffin embedded primary cancer tissue by immunohistochemistry and an immuno-reactive score (IRS) determined. Pathological remission was scored according to Sinn and by evaluation of cytopathic effects. NNAT-IRS was correlated with clinicopathological …parameters as well as relapse free and overall survival and for pathological remission after neoadjuvant therapy. RESULTS: NNAT IRS was an independent prognostic marker for relapse free and overall survival and the time from diagnosis to the “tumor-free” state. NNAT IRS was associated with Luminal-A tumors and correlated slightly negative with age and lymph-node metastasis. There was no significant correlation of NNAT-IRS with Sinn’s remission score, but with cytopathic effects of chemotherapy. CONCLUSIONS: We confirmed the prognostic impact of NNAT-IRS in an independent cohort of neoadjuvantly treated patients. Additionally, a correlation with a score for pathological remission under systemic neoadjuvant chemotherapy for breast cancer was found. Show more

Keywords: Breast cancer, neoadjuvant chemotherapy, neuronatin, remission score

DOI: 10.3233/CBM-203127

Citation: Cancer Biomarkers, vol. 32, no. 2, pp. 161-173, 2021

Authors: Seo, Danbi | Roh, Jungwook | Chae, Yeonsoo | Kim, Wanyeon

Article Type: Research Article

Abstract: Lung cancer accounts for a large proportion of cancer-related deaths worldwide. Personalized therapeutic medicine based on the genetic characteristics of non-small cell lung cancer (NSCLC) is a promising field, and discovering clinically applicable biomarkers of NSCLC is required. LINC00472 is a long non-coding RNA and has been recently suggested to be a biomarker of NSCLC, but little is known of its mechanism in NSCLC. Thus, the current study was performed to document changes in gene expression after LINC00472 overexpression in NSCLC cells. As a result of cell viability and migration assay, LINC00472 downregulated cell survival, proliferation, and motility. Transcriptome sequencing …analysis showed 3,782 genes expression were changed in LINC00472 overexpressing cells. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed most genes were associated with intracellular metabolism. The PPP1R12B, RGS5, RBM5, RBL2, LDLR and PTPRM genes were upregulated by LINC00472 overexpression and these genes functioned as tumor suppressors in several cancers. In contrast, SPSB1, PCNA, CD24, CDK5, CDC25A, and EIF4EBP1 were downregulated by LINC00472, and they functioned as oncogenes in various cancers. Consequently, the function of LINC00472 in tumorigenesis might be related to changes in the expressions of other oncogenes and tumor suppressors. Show more

Keywords: Long non-coding RNA LINC00472, transcriptome analysis, lung tumorigenesis

DOI: 10.3233/CBM-210242

Citation: Cancer Biomarkers, vol. 32, no. 2, pp. 175-188, 2021

Authors: Ma, Xiang | Wang, Younan | Fan, Hao | Zhu, Chuming | Chen, Wangwang | Li, Zengliang | Xiao, Jian | Ni, Peidong | Xu, Zekuan | Yang, Li

Article Type: Research Article

Abstract: BACKGROUND: Genetic polymorphisms are believed to represent a key aspect of predisposition to gastric cancer (GC). Therefore, considering the important role of Cathepsin B (CTSB) in promoting cancer onset and development, it could be very worthful to explore the function of CTSB-related genetic polymorphisms in GC. OBJECTIVE: In this study, we investigated the correlation of CTSB-related polymorphisms (rs9009A>T, rs6731T>C, rs1293303G>C, rs1874547C>T, rs3779659C>T, rs17814426C>T and rs148669985C>T) with GC risk and prognosis in a case-control study of 994 cases and 1000 controls. METHODS: All tag single nucleotide polymorphisms (SNPs) were genotyped by polymerase chain reaction-ligase detection …reaction (PCR-LDR) sequencing technology. RESULTS: The results indicated rs9009, rs6731 and rs17814426 correlated with decreased risks of GC (HR = 0.97, p < 0.001; HR = 0.86, P = 0.019; HR = 0.85, P = 0.017; respectively). Stratification analysis further showed rs17814426 variant genotypes correlated with earlier T stage (p = 0.044). In addition, GC patients carrying the C allele of rs6371 had better overall prognosis (HR = 0.62, 95%CI = 0.44–0.88). CONCLUSION: Our results firstly suggested the importance of CTSB-related polymorphisms on GC which could predict GC risk and prognosis. Show more

Keywords: Gastric cancer, Cathepsin B, Polymorphism, Susceptibility, Prognosis

DOI: 10.3233/CBM-203208

Citation: Cancer Biomarkers, vol. 32, no. 2, pp. 189-198, 2021

Authors: Zong, Rui | Chen, Xiaoman | Feng, Jingjing | Xu, Shan

Article Type: Research Article

Abstract: PURPOSE: Insulin like growth factor receptor 1 (IGF-1R) has been documented to play a key role in radiation response, thereby offering an attractive drug target to enhance tumor sensitivity to radiotherapy. Here, we investigated wether knockdown of IGF-1R can sensitize colorectal cancer (CRC) cell lines to radiation. MATERIAL AND METHODS: Human colon carcinoma SW480 and HT-29 cells were transfected with specific small interference RNA (siRNA) to mediate IGF-1R depletion. The expression of IGF-1R mRNA and protein among transfected and untransfected cells was detected by Western blot analysis. Changes in cell proliferation and radiosensitivity were evaluated by …the clonogenic survival assay. NVP-ADW742, an IGF-1R inhibitor, in combination with radiation was studied. RAD51, a measure for homologous recombination repair, and 53BP1, a maker for non-homologous end-joining (NHEJ), were determined by immunofluorescence for double-strand breaks (DSB) repair pathways. Cell cycle was also examined in the IGF-1R knockdown and IGF-1R-inhibited cells. RESULTS: CRC cell lines were selectively sensitized to radiation after siRNA-mediated IGF-1R depletion. NVP-ADW742 efficiently increases cancer cell response to radiation. Furthermore, initial formation of RAD51 foci after IR, and 53BP1 foci were significantly reduced in IGF-1R-depleted or with IGF-1R Inhibitor CRC cell lines. Lastly, IGF-1R-depleted or with IGF-1R Inhibitor caused more G2 phase cell arrest. CONCLUSION: Our findings demonstrate that depletion of IGF-1R lead to an increase in radiosensitivity in CRC. Show more

Keywords: Colorectal cancer, IGF-1R, neoadjuvant radiotherapy, radiosensitivity

DOI: 10.3233/CBM-210016

Citation: Cancer Biomarkers, vol. 32, no. 2, pp. 199-206, 2021

Authors: Zhang, Yi | Shao, Xuan | Gao, Chenyi | Xu, Danying | Wu, Jun | Zhu, Xuan | Chen, Zhigang

Article Type: Research Article

Abstract: BACKGROUND: FAS can serve as both an oncogene and a suppresser in different malignancies, and the prognostic value of FAS remains controversial. METHODS: The Oncomine database, KM-Plotter and bc-GenExMiner platform were adopted to analyze the prognostic value of FAS in breast cancer. Breast cancer tissue microarrays were further used to verify these data. The Cell Miner Tool was used to predict the value of FAS mRNA expression in predicting the efficacies of clinical drugs. RESULTS: We found that both FAS mRNA and protein expression level significantly reduced in breast carcinoma. In addition, high …FAS expression indicates a better metastatic relapse-free survival. Interestingly, FAS was associated with a better prognosis in different subtypes of breast cancer patients, namely, only in grade II and III, lymph nodal positive or p53 wild-type patients. The data from the Cell Miner Tool revealed that FAS mRNA expression was correlated with the efficacy of the first-line chemotherapeutic taxane agents and target drugs including olaparib and everolimus. CONCLUSIONS: FAS expression correlates with a better prognosis in breast cancer and may provide an effective clinical strategy to predict the sensitivity of taxanes and targeted drugs. Show more

Keywords: FAS, breast cancer, prognosis, efficacy of drugs

DOI: 10.3233/CBM-203125

Citation: Cancer Biomarkers, vol. 32, no. 2, pp. 207-219, 2021

Authors: Wang, Yu | Zhao, Han | Zhao, Ping | Wang, Xingang

Article Type: Research Article

Abstract: BACKGROUND: Pyruvate kinase M2 (PKM2) was overexpressed in many cancers, and high PKM2 expression was related with poor prognosis and chemoresistance. OBJECTIVE: We investigated the expression of PKM2 in breast cancer and analyzed the relation of PKM2 expression with chemotherapy resistance to the neoadjuvant chemotherapy (NAC). We also investigated whether PKM2 could reverse chemoresistance in breast cancer cells in vitro and in vivo . METHODS: Immunohistochemistry (IHC) was performed in 130 surgical resected breast cancer tissues. 78 core needle biopsies were collected from breast cancer patients before neoadjuvant chemotherapy. The relation of …PKM2 expression and multi-drug resistance to NAC was compared. The effect of PKM2 silencing or overexpression on Doxorubicin (DOX) sensitivity in the MCF-7 cells in vitro and in vivo was compared. RESULTS: PKM2 was intensively expressed in breast cancer tissues compared to adjacent normal tissues. In addition, high expression of PKM2 was associated with poor prognosis in breast cancer patients. The NAC patients with high PKM2 expression had short survival. PKM2 was an independent prognostic predictor for surgical resected breast cancer and NAC patients. High PKM2 expression was correlated with neoadjuvant treatment resistance. High PKM2 expression significantly distinguished chemoresistant patients from chemosensitive patients. In vitro and in vivo knockdown of PKM2 expression decreases the resistance to DOX in breast cancer cells in vitro and tumors in vivo. CONCLUSION: PKM2 expression was associated with chemoresistance of breast cancers, and could be used to predict the chemosensitivity. Furthermore, targeting PKM2 could reverse chemoresistance, which provides an effective treatment methods for patients with breast cancer. Show more

Keywords: Breast cancer, chemotherapy, pyruvate kinase M2

DOI: 10.3233/CBM-210111

Citation: Cancer Biomarkers, vol. 32, no. 2, pp. 221-230, 2021

Authors: Kang, Ben | Lee, Hyun Seok | Jeon, Seong Woo | Park, Soo Yeun | Choi, Gyu Seog | Lee, Won Kee | Heo, Somi | Lee, Duk Hee | Kim, Dong Sun

Article Type: Research Article

Abstract: BACKGROUND: Colorectal cancer (CRC) is one of the leading causes of mortality and morbidity in the world. It is characterized by different pathways of carcinogenesis and is a heterogeneous disease with diverse molecular landscapes that reflect histopathological and clinical information. Changes in the DNA methylation status of colon epithelial cells have been identified as critical components in CRC development and appear to be emerging biomarkers for the early detection and prognosis of CRC. OBJECTIVE: To explore the underlying disease mechanisms and identify more effective biomarkers of CRC. METHODS: We compared the levels and …frequencies of DNA methylation in 11 genes (Alu, APC, DAPK, MGMT, MLH1, MINT1, MINT2, MINT31, p16, RGS6, and TFPI2 ) in colorectal cancer and its precursor adenomatous polyp with normal tissue of healthy subjects using pyrosequencing and then evaluated the clinical value of these genes. RESULTS: Aberrant methylation of Alu, MGMT, MINT2 , and TFPI2 genes was progressively accumulated during the normal-adenoma-carcinoma progression. Additionally, CGI methylation occurred either as an adenoma-associated event for APC, MLH1, MINT1, MINT31, p16 , and RGS6 or a tumor-associated event for DAPK . Moreover, relatively high levels and frequencies of DAPK , MGMT , and TFPI2 methylation were detected in the peritumoral nonmalignant mucosa of cancer patients in a field-cancerization manner, as compared to normal mucosa from healthy subjects. CONCLUSION: This study identified several biomarkers associated with the initiation and progression of CRC. As novel findings, they may have important clinical implications for CRC diagnostic and prognostic applications. Further large-scale studies are needed to confirm these findings. Show more

Keywords: Colon mucosa, normal-adenoma-carcinoma, DNA methylation, pyrosequencing, biomarkers

DOI: 10.3233/CBM-203259

Citation: Cancer Biomarkers, vol. 32, no. 2, pp. 231-236, 2021

Authors: Cao, Yuting | Li, Qiang | Liu, Huihui | He, Xianglei | Huang, Fang | Wang, Yigang

Article Type: Research Article

Abstract: Over the past decade, cancer immunotherapy, such as immune checkpoint inhibitors (ICRs), has attained considerable progresses in clinical practice. T-cell immunoglobulin and mucin domain-containing protein 3 (Tim-3) act as next ICRs, and originally function as a co-inhibitory receptor expressed on interferon (IFN)-γ producing CD4 + and CD8 + T-cells. Furthermore, Tim-3 has also been found to express on innate immune cells and several types of tumors, signifying the pivotal role that Tim-3 plays in chronic viral infections and cancer. In addition, Tim-3 and multiple ICRs are concurrently expressed and …regulated on dysfunctional or exhausted T-cells, leading to improved antitumor immune responses in preclinical or clinical cancer therapy through co-blockade of Tim-3 and other ICRs such as programmed cell death-1 (PD-1). In this review, the biological characteristics of Tim-3 and the function of Tim-3 in regulating tumorigenesis and inflammation have been summarized. The usage of a single blockade of Tim-3 or in combination with multiple immunotherapy regimens have drawn attention to antitumor potential as a target for immunotherapy. Show more

Keywords: Immune checkpoint receptor, Tim-3, cancer immunotherapy, tumorigenesis, immune blockade

DOI: 10.3233/CBM-210114

Citation: Cancer Biomarkers, vol. 32, no. 2, pp. 237-248, 2021

Article Type: Correction

DOI: 10.3233/CBM-219528

Citation: Cancer Biomarkers, vol. 32, no. 2, pp. 249-, 2021