Cancer Biomarkers - Volume 35, issue 1

Authors: Takizawa, Satoko | Matsuzaki, Juntaro | Ochiya, Takahiro

Article Type: Review Article

Abstract: Circulating microRNA (miRNA) is a major focus in liquid biopsy studies. The circulating levels of certain miRNAs have been suggested to reflect specific physiological conditions, and several studies have reported their potential use as biomarkers for the detection and prognosis of cancer, as well as for predicting responses to chemotherapy or radiotherapy. Alongside these biomarker studies, research into the effects of specific background factors on circulating miRNA levels is progressing. Indeed, several studies have shown that a number of factors, including blood sample collection and processing methods, as well as subject-specific factors such as age, sex, and other physiological conditions, …can affect the normal levels of circulating miRNAs. Unfortunately, the evidence supporting these effects is not yet strong enough to support a definite conclusion and further research is warranted. Here, we summarize the findings of several studies that have addressed these concerns and identify important topics that should be considered when analyzing circulating miRNA levels in liquid biopsy studies. Show more

Keywords: miRNA, biomarker, cancer, serum, plasma

DOI: 10.3233/CBM-210223

Citation: Cancer Biomarkers, vol. 35, no. 1, pp. 1-9, 2022

Authors: Shi, Mingyue | Guo, Honggang | Bai, Yanliang | Niu, Junwei | Niu, Xiaona | Sun, Kai | Chen, Yuqing

Article Type: Research Article

Abstract: BACKGROUND: Mitosis-associated genes are dysregulated in many types of cancers and play important roles in disease progression and chemotherapy resistance. However, their expression and functions in chemotherapy-resistant Acute Myeloid Leukemia (AML) are still largely undetermined. OBJECTIVE: This study aims to explore the roles of spindle assembly checkpoint (SAC) genes CENPE, CENPF, and DLGAP5 in chemotherapy-resistant AML. METHODS: RNA-sequencing (RNA-seq) was performed in patients with chemotherapy-resistant AML and chemotherapy-sensitive AML. AML mRNA data from 151 patients with recurrence were downloaded from TCGA. Integrated analysis of the differentially expressed genes (DEGs), GO and KEGG pathways. …CENPE, CENPF, or DLGAP5 knockdown cell lines were used to analyse proliferation, apoptosis and cell cycle alterations. RESULTS: A total of 87 DEGs (48 upregulated and 39 downregulated) were obtained through gene analysis of R/R-AML and a total of 329 DEGs (202 upregulated and 127 downregulated) were obtained in refractory S-AML. Upregulated DEGs were mainly enriched in cell cycle (GO: 0007049, hsa04110) and mitotic cell cycle (GO: 0000278) processes and pathway. Venn diagram analysis identified the most upregulated DEGs (including CENPE, CENPF, and DLGAP5) in chemoresistant AML. The expression of CENPE, CENPF and DLGAP5 in R-AML (TCGA) was significantly higher than that of primary AML (GEO). The proliferation of K562 cells after CENPE and DLGAP5 knockdown was significantly decreased (P = 0.0001 and P = 0.0006). In THP-1 cells, the CCK-8 values after CENPE, CENPF and DLGAP5 knockdown were significantly decreased (P = 0.01, P = 0.0395 and P = 0.0362). Knockdown of CENPE, CENPF and DLGAP5 significantly increased cell apoptosis by regulating Caspase-9, BAX, TP-53 and bcl-2, and induced cell cycle arrested by regulating CDK1, CDK2, CDKN1A, and CyclinD1. CONCLUSIONS: In conclusion, the mitotic cell cycle-associated genes CENPE, CENPF, and DLGAP5 were upregulated in chemotherapy-resistant AML patients and might be useful for predicting poor prognosis. Show more

Keywords: Relapsed, refractory, AML, prognosis, CENPE, CENPF, DLGAP5

DOI: 10.3233/CBM-203170

Citation: Cancer Biomarkers, vol. 35, no. 1, pp. 11-25, 2022

Authors: Safaei, Sadegh | Sajed, Roya | Saeednejad Zanjani, Leili | Rahimi, Mandana | Fattahi, Fahimeh | Ensieh Kazemi-Sefat, Golnaz | Razmi, Mahdieh | Dorafshan, Shima | Eini, Leila | Madjd, Zahra | Ghods, Roya

Article Type: Research Article

Abstract: BACKGROUND: Dynamin 2 (DNM2) involved in tumor progression in various malignancies. OBJECTIVE: For the first time, we evaluated DNM2 expression pattern, its association with clinicopathological characteristics and survival outcomes in RCC subtypes. METHODS: We evaluated the DNM2 expression pattern in RCC tissues as well as adjacent normal tissue using immunohistochemistry on tissue microarray (TMA) slides. RESULTS: Our findings revealed increased DNM2 expression in RCC samples rather than in adjacent normal tissues. The results indicated that there was a statistically significant difference between cytoplasmic expression of DNM2 among subtypes of RCC …in terms of intensity of staining, percentage of positive tumor cells, and H-score (P = 0.024, 0.049, and 0.009, respectively). The analysis revealed that increased cytoplasmic expression of DNM2 in ccRCC is associated with worse OS (log rank: P = 0.045), DSS (P = 0.049), and PFS (P = 0.041). Furthermore, cytoplasmic expression of DNM2 was found as an independent prognostic factor affecting DSS and PFS in multivariate analysis. CONCLUSIONS: Our results indicated that DNM2 cytoplasmic expression is associated with tumor aggressiveness and poor outcomes. DNM2 could serve as a promising prognostic biomarker and therapeutic target in patients with ccRCC. Show more

Keywords: DNM2, renal cell carcinoma (RCC), ccRCC, immunohistochemistry (IHC), tissue microarray (TMA)

DOI: 10.3233/CBM-210514

Citation: Cancer Biomarkers, vol. 35, no. 1, pp. 27-45, 2022

Authors: Contreras-Sanzón, Estefania | Palma-Flores, Carlos | Flores-Pérez, Ali | M. Salinas-Vera, Yarely | B. Silva-Cázares, Macrina | A. Marchat, Laurence | G. Avila-Bonilla, Rodolfo | N. Hernández de la Cruz, Olga | E. Álvarez-Sánchez, María | Pérez-Plasencia, Carlos | D. Campos-Parra, Alma | López-Camarillo, César

Article Type: Research Article

Abstract: BACKGROUND: Vasculogenic mimicry (VM) is characterized by formation of three-dimensional (3D) channels-like structures by tumor cells, supplying the nutrients needed for tumor growth. VM is stimulated by hypoxic tumor microenvironment, and it has been associated with increased metastasis and clinical poor outcome in cancer patients. cAMP responsive element (CRE)-binding protein 5 (CREB5) is a hypoxia-activated transcription factor involved in tumorigenesis. However, CREB5 functions in VM and if its regulated by microRNAs remains unknown in breast cancer. OBJECTIVE: We aim to study the functional relationships between VM, CREB5 and microRNA-204-5p (miR-204) in breast cancer cells. …METHODS: CREB5 expression was evaluated by mining the public databases, and using RT-qPCR and Western blot assays. CREB5 expression was silenced using short-hairpin RNAs in MDA-MB-231 and MCF-7 breast cancer cells. VM formation was analyzed using matrigel-based cultures in hypoxic conditions. MiR-204 expression was restored in cancer cells by transfection of RNA mimics. Luciferase reporter assays were performed to evaluate the binding of miR-204 to 3 ′ UTR of CREB5. RESULTS: Our data showed that CREB5 mRNA expression was upregulated in a set of breast cancer cell lines and clinical tumors, and it was positively associated with poor prognosis in lymph nodes positive and grade 3 basal breast cancer patients. Silencing of CREB5 impaired the hypoxia-induced formation of 3D channels-like structures representative of the early stages of VM in MDA-MB-231 cells. In contrast, VM formation was not observed in MCF-7 cells. Interestingly, we found that CREB5 expression was negatively regulated by miR-204 mimics in breast cancer cells. Functional analysis confirmed that miR-204 binds to CREB5 3 ′ -UTR indicating that it’s an ulterior effector. CONCLUSIONS: Our findings suggested that CREB5 could be a potential biomarker of disease progression in basal subtype of breast cancer, and that perturbations of the miR-204/CREB5 axis plays an important role in VM development in breast cancer cells. Show more

Keywords: Breast cancer, microRNA-204-5p, CREB5, prognosis, vasculogenic mimicry

DOI: 10.3233/CBM-210457

Citation: Cancer Biomarkers, vol. 35, no. 1, pp. 47-56, 2022

Authors: Tcherkassova, Janneta | Prostyakova, Anna | Tsurkan, Sergey | Ragoulin, Vladislav | Boroda, Alexander | Sekacheva, Marina

Article Type: Research Article

Abstract: BACKGROUND: Combination of different cancer markers is often used for predicting tumor growth, for the response to cancer therapy, and for increase in the positive diagnosis ratio in the malignant tumors. OBJECTIVE: Evaluation of the diagnostic efficacy of CA 15-3 and CA-62 cancer markers combination for early stages of breast cancer (BC) detection. METHODS: This blind study was performed on 2 clinically validated Sets that included serum measurements of CA 15-3 ELISA and CLIA-CA-62 assays in 488 serum samples with TNM classification. A study included 300 BC patients (254 at Stages I and …II, 20 with ductal carcinoma in situ (DCIS), and 26 Stages III and IV patients), 47 patients with breast benign diseases, and 141 healthy controls. RESULTS: Sensitivity for DCIS & Stage I breast cancer detection was 75% at 100% Specificity (AUC = 0.895) using a following combination of two antigens: 10 < CA15-3 < 46 U/ml and CA-62 ⩾ 6300 U/ml, which allows eliminating false positive results. CONCLUSIONS: The results obtained in a blind study demonstrate that a combination of CA15-3 with CA-62 yields 75% Sensitivity at 100% Specificity for DCIS and Stage I breast cancer detection, which has a potential to be integrated into existing screening programs. Show more

Keywords: Breast cancer (BC), Stage I, CA 15-3, CA-62, cancer marker

DOI: 10.3233/CBM-210533

Citation: Cancer Biomarkers, vol. 35, no. 1, pp. 57-69, 2022

Authors: Li, Liang | Gao, Jie | Li, Jiangang | Wang, Jun

Article Type: Research Article

Abstract: BACKGROUND: MicroRNAs (miRNAs) have been reported to play an important role in tumor progression by regulating the expression of target genes. OBJECTIVE: This study attempted to verify the role of miR-711 in gastric cancer (GC) progression by in vitro and in vivo assays. METHODS: The expression of miR-711 in tumor tissues and cells was detected by real-time quantitative PCR (qRT-PCR). Expression of MiR-711 in NCI-N87 and SNU-1 cells was detected by FISH. We transfected GC cells with miR-711 mimics or inhibitors. The effects of miR-711 on the proliferation and metastasis of …GC cells were detected by CCK-8, wound healing and transwell assays. Dual-luciferase reporter gene assay was used to verify the targeting relationship between miR-711 and CD44. Xenograft assays was used to verify the regulatory effect of miR-711 on tumor growth. RESULTS: In GC tissues and cell lines, the expression of miR-711 was down-regulated when compare with adjacent tissues or normal epithelial cells. The results indicated that overexpressing of miR-711 could suppress the GC cell proliferation, migration, and invasion through targeting CD44. The knockdown of CD44 showed similar effects as miR-711 overexpression in GC cells. Moreover, we confirmed these effects in the in vivo assays. Furthermore, we found that miR-711 could play a role by influencing tumor cell stemness. CONCLUSION: MiR-711 plays vital roles as a tumor-suppressor by targeting CD44 and may be a therapeutic target for GC treatment. Show more

Keywords: Gastric cancer, MiR-711, cancer stem cells, CD44, MicroRNAs

DOI: 10.3233/CBM-210213

Citation: Cancer Biomarkers, vol. 35, no. 1, pp. 71-81, 2022

Authors: Lian, Zhenpeng | Chang, Taihao | Ma, Shenfei | Li, Jing | Zhang, Hongtuan | Wang, Xiaoming | Liu, Ranlu

Article Type: Research Article

Abstract: OBJECTIVE: The N-myc downstream-regulated gene 1 (NDRG1) has been discovered as a significant gene in the progression of cancers. However, the regulatory mechanism of NDRG1 remained obscure in prostate cancer (PCa). METHODS: The miR-96-5p and NDRG1 expression levels were evaluated in PCa cell lines, prostate tissues, and validated public databases by real-time PCR, western blot analysis, and immunohistochemistry. The function of miR-96-5p and NDRG1 were investigated by wound healing and transwell assays in vitro , and mouse xenograft assay in vivo . The candidate pathway regulated by NDRG1 was conducted by the next-generation gene sequencing technique. …Immunofluorescence and luciferase assay was used to detect the relation between miR-96-5p, NDRG1, and NF-κ B pathway. RESULTS: Overexpressing NDRG1 suppresses the migration, invasion, and epithelial-mesenchymal transition (EMT) in vitro , and inhibits metastasis in vivo . Moreover, miR-96-5p contributes to NDRG1 deficiency and promotes PCa cell migration and invasion. Furthermore, NDRG1 loss activates the NF-κ B pathway, which stimulates p65 and IKBa phosphorylation and induces EMT in PCa. CONCLUSIONS: MiR-96-5p promotes the migration and invasion of PCa by targeting NDRG1 and regulating the NF-κ B pathway. Show more

Keywords: Prostate cancer, NDRG1, miR-96-5p, NF-B, EMT

DOI: 10.3233/CBM-210072

Citation: Cancer Biomarkers, vol. 35, no. 1, pp. 83-98, 2022

Authors: Lee, Dae Hoon | Paik, E. Sun | Cho, Young-Jae | Lee, Yoo-Young | Lee, Bada | Lee, Eui Jin | Choi, Jung-Joo | Choi, Chel-Hun | Lee, Sangmin | Choi, Jin Woo | Lee, Jeong-Won

Article Type: Research Article

Abstract: BACKGROUND: Although lysyl-tRNA synthetase (KARS1) is predominantly located in the cytosol, it is also present in the plasma membrane where it stabilizes the 67-kDa laminin receptor (67LR). This physical interaction is strongly increased under metastatic conditions. However, the dynamic interaction of these two proteins and the turnover of KARS1 in the plasma membrane has not previously been investigated. OBJECTIVE: Our objective in this study was to identify the membranous location of KARS1 and 67LR and investigate if this changes with the developmental stage of epithelial ovarian cancer (EOC) and treatment with the inhibitor BC-K01. In addition, …we evaluated the therapeutic efficacy of BC-K01 in combination with paclitaxel, as the latter is frequently used to treat patients with EOC. METHODS: Overall survival and prognostic significance were determined in EOC patients according to KARS1 and 67LR expression levels as determined by immunohistochemistry. Changes in the location and expression of KARS1 and 67LR were investigated in vitro after BC-K01 treatment. The effects of this compound on tumor growth and apoptosis were evaluated both in vitro and in vivo . RESULTS: EOC patients with high KARS1 and high 67LR expression had lower progression-free survival rates than those with low expression levels of these two markers. BC-K01 reduced cell viability and increased apoptosis in combination with paclitaxel in EOC cell xenograft mouse models. BC-K01 decreased membranous KARS1 expression, causing a reduction in 67LR membrane expression in EOC cell lines. BC-K01 significantly decreased in vivo tumor weight and number of nodules, especially when used in combination with paclitaxel. CONCLUSIONS: Co-localization of KARS1 and 67LR in the plasma membrane contributes to EOC progression. Inhibition of the KARS1-67LR interaction by BC-K01 suppresses metastasis in EOC. Show more

Keywords: Laminin receptor, lysyl-tRNA synthetase, cancer metastasis, anticancer agent, protein-protein interaction

DOI: 10.3233/CBM-210077

Citation: Cancer Biomarkers, vol. 35, no. 1, pp. 99-109, 2022

Authors: Zeng, Yufei | Gao, Weiqi | Chen, Xiaosong | Shen, Kunwei

Article Type: Research Article

Abstract: BACKGROUND: The 21-gene recurrence score (RS) assay has been validated to predict the risk of disease-free survival in estrogen receptor (ER)-positive, human epidermal growth factor receptor 2 (HER2)-negative early breast cancer patients. However, its relation with locoregional recurrence (LRR) risk is unclear. OBJECTIVE: This study aimed to explore the ability of RS to predict LRR risk. METHODS: Consecutive ER-positive, HER2-negative, pT1, pN0-1, and M0 early breast cancer patients were analyzed retrospectively. According to RS, patients were divided into low- (RS < 18), intermediate- (RS 18–30), and high-risk (RS ⩾ …31) groups. The primary endpoint was LRR. Subgroup analysis was conducted according to different nodal statuses and surgery types. RESULTS: A total of 1558 patients were enrolled: 354 (22.7%), 788 (50.6%), and 416 (26.7%) patients in the low-, intermediate-, and high-risk groups, respectively. The median follow-up time was 53 months, and 2, 8, and 14 LRR events were observed in the low-, intermediate-, and high-risk groups, respectively (P = 0.004). Both univariate (P = 0.009) and multivariate (P = 0.010) analyses demonstrated that 21-gene RS was correlated with LRR. Compared to low-risk patients, high-risk patients were at greater risk of LRR (HR 5.75, 95% CI 1.30–25.31, P = 0.021). Among pN0 (P = 0.033), pN1 (P = 0.049) and postmastectomy patients (P = 0.012), 21-gene RS remained predictive of the risk of LRR. CONCLUSION: The 21-gene RS assay was significantly associated with the risk of LRR in ER-positive, HER2-negative early breast cancer patients. Among patients with different nodal statuses and patients receiving mastectomy, RS remained predictive of the risk of LRR. Show more

Keywords: 21-gene RS assay, breast cancer, locoregional recurrence, mastectomy, breast-conserving surgery

DOI: 10.3233/CBM-210274

Citation: Cancer Biomarkers, vol. 35, no. 1, pp. 111-118, 2022

Authors: Ding, Hao | Chen, Yuxing | Zhao, Yuanyang | Zhu, Li | Huang, Huaying | Liu, Chenyang | Zhang, Feng | Zhang, Cunxi | Jin, Cheng

Article Type: Research Article

Abstract: BACKGROUND: EGFR mutations widely exists in NSCLC patients, which are involved in cancer development. OBJECTIVE: The function of EGFR mutations in the resistance to TKI treatments of NSCLC was evaluated to provide theoretical support for the clinical management of NSCLC patients. METHODS: A total of 150 NSCLC patients including 118 patients with EGFR mutation and 32 without, were included in this study. The EGFR mutation status and subtypes were analyzed in recruited patients. The distribution of EGFR mutation subtypes and their association with clinicopathological features were also assessed. The prognostic value of EGFR …mutation was evaluated by the overall survival of recruited patients. The function of EGFR mutation was estimated, in vitro , in the TKI resistant NSCLC cells with different subtypes of EGFR mutation. RESULTS: The exon 19 deletion was the most common subtype of EGFR mutation in the enrolled patients followed by the exon 21 L858R point mutation. The EGFR mutations were closely associated with the differentiation degree and the histological types of NSCLC cases. EGFR mutation was an independent prognostic factor of NSCLC with a close relationship with the overall survival of patients. The exon 20 T790M mutation results in the erlotinib resistance through the PI3K/Akt signaling pathway. CONCLUSIONS: The EGFR mutation is a critical factor in the prognosis and for the resistance to TKI treatment in NSCLC. The exon 20 T790M mutation was involved in the erlotinib resistance through PI3K/Akt signaling pathway. Show more

Keywords: EGFR mutation, NSCLC, TKI-resistance, erlotinib, serum, exon 20

DOI: 10.3233/CBM-210281

Citation: Cancer Biomarkers, vol. 35, no. 1, pp. 119-125, 2022