Cancer Biomarkers - Volume 28, issue 1

Authors: Liu, Longyang | Yi, Juanjuan | Yuan, Jianhuan | Yao, Tingting | Lin, Zhongqiu | Ning, Yingxia | Zeng, Zhaoyang

Article Type: Research Article

Abstract: OBJECTIVE: To investigate FOXO1 expression in epithelial ovarian cancer (EOC), and to explore its correlation with clinicopathological parameters and prognosis of EOC. METHODS: Two hundred and sixteen cases of paraffin-embedded EOC and 41 paratumor tissues from 2009 to 2017 that had been pathologically confirmed at the memorial hospital of Sun Yat-sen University were included in this study, and the expression of FOXO1 was performed by immunohistochemistry using a polyclonal antibody specific for FOXO1. RESULTS: FOXO1 protein expression is associated with Recurrence free and overall survival in EOC patients; In addition, FOXO1 expression is …associated with age, FIGO stage, intraperitoneal metastasis, intestinal metastasis, vital status, intraperitoneal recurrence and differentiation grade; Moreover, in a multivariate model FOXO1 overexpression was an independent predictor of poor survival in EOC. CONCLUSION: FOXO1 may play a candidate oncogenic role in EOC, and FOXO1 is a useful independent prognostic marker in EOC, and it may provide a candidate target therapy in future. Show more

Keywords: FOXO1, epithelial ovarian cancer, prognosis

DOI: 10.3233/CBM-182119

Citation: Cancer Biomarkers, vol. 28, no. 1, pp. 1-8, 2020

Authors: Fang, Zheping | Wu, Linjun | Dai, Haojiang | Hu, Peng | Wang, Binfeng | Han, Qiuyue | Xu, Yongfu | Lv, Shangdong | Zhu, Yu | Gan, Meifu | Zhou, Weijie | Zhang, Wenlong

Article Type: Research Article

Abstract: BACKGROUND: Recently, hepatocellular carcinoma (HCC) has been ranked as the second leading cause of cancer-associated death. However, the underlying molecular mechanisms of HCC progression remain unclear. Vesicular overexpressed in cancer pro-survival protein 1 (VOPP1) could be upregulated in a quantity of human cancers, including squamous cell carcinoma (SCC), gastric cancer, and glioblastoma. However, the precise functional mechanism of VOPP1 in HCC remains poorly understood. The present study aimed to investigate the role of VOPP1 in HCC proliferation. METHODS: Immunohistochemistry (IHC), Western blot and Reverse-transcription polymerase chain reaction (RT-PCR) were used to analyze the protein and mRNA …expressions of VOPP1, mitogen-activated protein kinase (MAPK) 14, ribosomal protein S6 kinase β 1 (RPS6KB1), cylindromatosis (CYLD) and Twist family bHLH transcription factor 1 (TWIST1). The cell proliferation and apoptosis were tested using Celigo cell imaging analyzer and annexin V-APC apoptosis detection kit respectively. Colony formation and tumor xenograft assays were performed to understand their roles in tumorigenicity. RESULTS: The expression of VOPP1 in HCC samples was higher than that in adjacent noncancerous tissues by immunohistochemistry. In addition, the down-regulation of VOPP1 using shRNA inhibited cell proliferation and tumour growth, and induced cell apoptosis in vitro and in vivo . Furthermore, VOPP1 silencing increased the expression of MAPK14 and RPS6KB1, indicating that the MAPK and mTOR signalling pathways might be involved in VOPP1-mediated cancer cell proliferation. CONCLUSION: The present data indicate that VOPP1 may play an important role in the progression of HCC by targeting the MAPK and mTOR signalling pathways, and that VOPP1 may potentially be a candidate as a novel molecular target for HCC therapy. Show more

Keywords: VOPP1, hepatocellular carcinoma, proliferation, apoptosis

DOI: 10.3233/CBM-190574

Citation: Cancer Biomarkers, vol. 28, no. 1, pp. 9-20, 2020

Authors: Ling, Wang | Yangchun, Xu | Wei, Wang | Qiang, Wang

Article Type: Research Article

Abstract: INTRODUCTION: The present study evaluated the effects of the long non-coding RNA (lncRNA) gastric carcinoma high-expressed transcript (GHET1) in cervical carcinoma development. METHODS: The expression levels of GHET1 and PTEN were measured using in situ hybridisation, immunohistochemistry (IHC) and quantitative reverse transcription PCR assay to investigate their correlations. In an in vitro study, the effects of GHET1 knockdown on the biological activities of SiHa and HeLa cells were evaluated by MTT, flow cytometry, transwell and wound-healing assays and relative protein expression was measured using western blotting. In an in vivo experiment, cell apoptosis and …relative protein expression were measured in nude mice using TUNEL and IHC assays, respectively. RESULTS: The expression levels of lncRNA GHET1 and PTEN protein differed significantly between cancer and adjacent normal tissues (P < 0.05) and were negatively correlated in the clinical data. In vitro, proliferation rateswere significantly down-regulated in SiHa and HeLa cells. The GHET1 knockdown (si-GHET1) groups showed significantly higher G 1 phase and apoptosis rates and significantly suppressed invasion and migration abilities compared with the normal control (NC) group (P < 0.05 for all). The expression levels of PTEN, PI3 K, AKT, P53, matrix metalloproteinase (MMP)-2 and MMP-9 proteins differed significantly between the si-GHET1 and NC groups (P < 0.05 for all). In vitro, the lncRNA group showed significantly suppressed tumour volume and weight, increased cell apoptosis and different relative protein expression levels compared with the NC group (P < 0.05 for all). CONCLUSION: GHET1 knockdown suppressed cervical carcinoma development via the PTEN/PI3 K/AKT signalling pathway. Show more

Keywords: GHET1, PTEN, PI3 K, AKT, SiHa, HeLa

DOI: 10.3233/CBM-190269

Citation: Cancer Biomarkers, vol. 28, no. 1, pp. 21-32, 2020

Authors: Matsumoto, Kazumasa | Murakami, Yasukiyo | Shimizu, Yuriko | Hirayama, Takahiro | Ishikawa, Wataru | Iwamura, Masatsugu

Article Type: Research Article

Abstract: BACKGROUND: No study has yet investigated the use of electronic nose (eNose) technology to reveal pattern recognition of urological diseases, including bladder cancer. OBJECTIVE: We sought to determine the diagnostic performance of the eNose in recognizing urinary odour in patients with bladder cancer. METHODS: The eNose is a commercially available model equipped with two sensors. The angle of the two sensors (θ ) depends on the kinds of chemical substances, thus defining θ as the feature of odour. Quantity of odour is the number of θ …detected during a measurement. Urine samples were from 36 untreated patients with bladder cancer, 29 with urolithiasis, 10 with urinary tract infection (UTI), and 27 healthy volunteers. RESULTS: Based on ROC analysis of the quantity in patients with bladder cancer, an optimal cut-off value for θ of 49, 48, and 55 was applied to compare with samples from the healthy volunteer, urolithiasis and UTI groups, respectively. There were significantly differences between bladder cancer and the other conditions using these specific points (p < 0.0001, respectively). The resulting diagnostic sensitivity was 61.4%, 45.6%, and 60.8%, and specificity was 52.8%, 68.4%, and 90.2%, respectively. The AUC for bladder cancer was 0.565, 0.548, and 0.909, respectively. CONCLUSION: The eNose is a small, portable, rapid, low cost, and noninvasive instrument for distinguishing bladder cancer from other benign conditions. Show more

Keywords: Bladder cancer, electronic nose, urine, odour

DOI: 10.3233/CBM-190466

Citation: Cancer Biomarkers, vol. 28, no. 1, pp. 33-39, 2020

Authors: Li, Huixia | Guan, Congjin

Article Type: Research Article

Abstract: BACKGROUND: To investigate the influences of HOX transcript antisense ribonucleic acid (HOTAIR) on the proliferation and apoptosis of glioblastoma cells by targeting micro RNA (miR)-219. OBJECTIVE: With glioblastoma cell line U87 as the object, the changes in expression levels of HOTAIR and miR-219 in each group were detected via quantitative real-time polymerase chain reaction (qRT-PCR) after HOTAIR in U87 cell lines was knocked down using small interfering RNA (siRNA). Then the cell proliferation in each group was determined using Cell Counting Kit-8 (CCK-8) and colony formation assays. Flow cytometry was applied to detect the cell apoptosis, …and Western blotting assay was adopted to measure the changes in protein levels of Cyclin D1 and Bcl-2-associated X protein (Bax). After miR-219 knockdown with siRNA, the changes in expression levels of HOTAIR and miR-219 in each group were examined through qRT-PCR, and the cell proliferation was tested by CCK-8 assay. RESULTS: After interference inHOTAIR using siRNA, compared with those in control group, the RNA expression level of HOTAIR was decreased remarkably (p < 0.05), the RNA expression level of miR-219 was increased notably (p < 0.05), the cell proliferation rate was inhibited evidently (p < 0.05), the apoptosis rate was enhanced obviously (p < 0.05), the protein expression level of Cyclin D1 declined markedly (p < 0.05), and the protein expression level of Bax rose distinctly (p < 0.05) in HOTAIR-siRNA group. After miR-219 knockdown with siRNA, the cell proliferation rate was raised remarkably (p < 0.05), but there was no significant change in the RNA expression level of HOTAIR (p > 0.05). CONCLUSION: HOTAIR can repress the proliferation and promote the apoptosis of glioblastoma cells by targeting miR-219. Show more

Keywords: HOTAIR, miR-219, glioblastoma, proliferation and apoptosis

DOI: 10.3233/CBM-190467

Citation: Cancer Biomarkers, vol. 28, no. 1, pp. 41-47, 2020

Authors: Ali, Marwa A. | Shaker, Olfat G. | Alazrak, Mohammed | AbdelHafez, Marwa N. | Khalefa, Abeer A. | Hemeda, Nada F. | Abdelmoktader, Abdelrahman | Ahmed, Fatma A.

Article Type: Research Article

Abstract: BACKGROUND: LncRNA MEG3 rs7158663 has been shown to confer cancer susceptibility, maybe through altering its gene expression level. OBJECTIVE: We aimed to weigh the effect of rs7158663 on MEG3 serum level and breast cancer susceptibility. METHODS: We genotyped rs7158663 G > A and measured serum MEG3 in 150 breast cancer, 95 fibroadenoma , and 154 controls by the TaqMan method. RESULTS: The presence of rs7158663 G > A is a risk factor for breast cancer among fibroadenoma patients and controls, AA vs. GG genotypes …(OR = 6.320, 95% CI = 2.587–15.439, P < 0.0001 when compared to controls and OR = 10.825, 95% CI = 1.929–60.742, P = 0.007 when compared to fibroadenoma). Decreased serum MEG3 was observed in breast cancer group when compared with fibroadenoma and/or controls [median (IQR) = 0.43 (0.27–0.55)] (P < 0.0001). However, increased serum MEG3 was noted in fibroadenoma group when compared with controls (P < 0.0001). A significance decreased serum MEG3 was found to be associated with mutant A allele than with wild G allele (P < 0.0001). The results showed that rs7158663 and lower MEG3 were significantly associated with patients with higher TNM staging and larger tumor size > 5 cm. CONCLUSION: The presence of both rs7158663 and low MEG3 are diagnostic and unfavorable prognostic factors for BC patients. Show more

Keywords: MEG3, polymorphism, rs7158663, breast cancer, fibroadenoma

DOI: 10.3233/CBM-191072

Citation: Cancer Biomarkers, vol. 28, no. 1, pp. 49-63, 2020

Authors: Zeng, Zhaoyang | Ji, Na | Yi, Juanjuan | Lv, Jin | Yuan, Jianhuan | Lin, Zhongqiu | Liu, Longyang | Feng, Xin

Article Type: Research Article

Abstract: OBJECTIVE: LGR4 expression in serous ovarian cancer paraffin-embedded tissues and fresh tissues were investigated, and its expression associated with clinicopathological parameters and prognosis in serous ovarian cancer was explored. METHODS: From Dec, 2009 to Jan, 2020, 122 paraffin-embedded serous ovarian cancer patients and 41 paired paratumor tissues who were both diagnosed and operated at the memorial hospital of Sun Yat-sen University and Integrated Hospital of Traditional Chinese Medicine, Southern Medical University were selected in this research, respectively, and all of these tissues were performed by immunohistochemistry (IHC) with a polyclonal antibody for LGR4. Meanwhile, from Aug, …2013 to Mar, 2019, 15 cases of serous ovarian cancer fresh tissues and 15 cases of paratumor fresh tissues who were operated at Integrated Hospital of Traditional Chinese Medicine, Southern Medical University were performed with Quantitative Real-time PCR to detect the mRNA expression of LGR4, respectively. RESULTS: LGR4 expression was much higher both in paraffin-embedded and fresh cancer tissues than that in paratumor tissues, respectively, and its expression was associated with recurrence free survival and overall survival in serous ovarian cancer patients. Moreover, in a multivariate model LGR4 was an indeed independent predictor of poor survival in serous ovarian cancer patients. CONCLUSION: LGR4 is upregulated in serous ovarian cancer, and LGR4 is an indeed useful independent prognostic predictor in serous ovarian cancer, and it may provide important clinical value of serous ovarian cancer. Show more

Keywords: LGR4, serous ovarian cancer, prognosis

DOI: 10.3233/CBM-191145

Citation: Cancer Biomarkers, vol. 28, no. 1, pp. 65-72, 2020

Authors: Yang, Junsheng | Zhang, Zhen | Chen, Shaoping | Dou, Wenwen | Xie, Ruizhu | Gao, Jie

Article Type: Research Article

Abstract: BACKGROUND: Increasing evidence reveals that aberrant microRNAs (miRNAs) expression play a crucial role in the tumorigenesis of cancers, including hepatocellular carcinoma (HCC), whereas the role of miR-654-3p in HCC remains unclear. This study aimed to investigate the role of miR-654-3p in HCC. METHODS: Real-time quantitative PCR was performed to detect miR-654-3p expression in HCC tissues and cell lines. The association of miR-654-3p expression with clinical characteristics of HCC patients were analyzed. And the prognostic value of miR-654-3p was examined using Kaplan-Meier curve and Cox regression analysis. CCK-8 and Transwell assays were used to observe the effects …of miR-654-3p on proliferation, migration, and invasion of HCC cells. RESULTS: The miR-654-3p expression was downregulated in both HCC tissues and cell lines, which was significantly associated with lymph node metastasis and TNM stage. Downregulation of miR-654-3p predicted poor prognosis of HCC patients. Overexpression of miR-654-3p inhibited HCC cell proliferation, migration, and invasion, while knockdown of miR-654-3p promoted these cellular behaviors in vitro . CONCLUSION: Our study suggested that miR-654-3p expression was downregulated in HCC and might serve as a potential prognostic marker and therapeutic target for the survival of HCC patients. miR-654-3p might exert a suppressor role in HCC through inhibiting tumor cell proliferation, migration, and invasion. Show more

Keywords: Hepatocellular carcinoma, invasion, miR-654-3p, migration, prognosis, proliferation

DOI: 10.3233/CBM-191084

Citation: Cancer Biomarkers, vol. 28, no. 1, pp. 73-79, 2020

Authors: Hyun, Jae-Won | Shin, Hyung Shik | Kim, Su-Hyun | Kong, Sun-Young | Yoo, Heon | Gwak, Ho-Shin | Kim, Ho Jin

Article Type: Research Article

Abstract: BACKGROUND: To investigate the feasibility of cerebrospinal fluid (CSF) CYFRA 21-1 levels as a therapeutic monitoring biomarker in leptomeningeal carcinomatosis (LMC) patients undergoing ventriculo-lumbar perfusion (VLP) chemotherapy. METHODS: The levels of CYFRA 21-1 in 42 CSF samples from 15 LMC patients were analyzed using an electrochemiluminescence immunoassay. Samples were collected at individual time points during VLP chemotherapy. Therapeutic outcomes were measured as improvements in the Karnofsky Performance Status (KPS) score and decreasing intracranial pressure (ICP) as the main endpoint of VLP chemotherapy. Changes in CSF CYFRA 21-1 levels, protein levels, and cytology results were also investigated. …We subsequently evaluated whether these changes were correlated with KPS score and ICP. RESULTS: The CSF CYFRA 21-1 levels at individual time points were associated with KPS score and ICP. The KPS scores (p = 0.007) and ICP (p = 0.018) of patients with high CSF CYFRA 21-1 levels were significantly different from those of patients with low CSF CYFRA 21-1 levels. By contrast, CSF protein levels and cytological responses were not significantly associated with KPS scores and ICP. CONCLUSIONS: CSF CYFRA 21-1 may have utility as a therapeutic monitoring biomarker to design personalized therapeutic strategies in LMC patients undergoing VLP chemotherapy. Show more

Keywords: Leptomeningeal metastasis, monitoring, biomarker, CSF, CYFRA 21-1

DOI: 10.3233/CBM-190187

Citation: Cancer Biomarkers, vol. 28, no. 1, pp. 81-89, 2020

Authors: Kaikkonen, Elina | Takala, Aliisa | Pursiheimo, Juha-Pekka | Wahlström, Gudrun | Schleutker, Johanna

Article Type: Research Article

Abstract: BACKGROUND: Elevated Anoctamin 7 (ANO7 ) expression is associated with poor survival in prostate cancer patients. OBJECTIVE: The aim was to discover proteins that interact with ANO7 to understand its functions and regulatory mechanisms. METHODS: The proximity-dependent biotin identification (BioID) method was utilized. ANO7 fused to biotin ligase was transiently transfected into LNCaP cells, and the biotinylated proteins were collected and analysed by mass spectrometry. Four identified proteins were stained with dual fluorescent immunostaining and visualized using Stimulated emission depletion microscopy (STED). RESULTS: After bioinformatic filtering steps, 64 potentially ANO7-interacting …proteins were identified and analysed with the GO enrichment analysis tool. One of the most prominently enriched cellular components was cellular vesicle. Co-localization was showed for staphylococcal nuclease and tudor domain containing 1 (SND1), heat shock protein family A (Hsp70) member 1A (HSPA1A), adaptor related protein complex 2 subunit beta 1 (AP2B1) and coatomer protein complex subunit gamma 2 (COPG2). CONCLUSIONS: This is the first study in which ANO7 interacting proteins have been identified. Although further studies are needed, the findings reported here expand our understanding of the role and regulation of ANO7 in prostate cancer cells. Furthermore, these results are likely to introduce new targets for the novel cancer therapies. Show more

Keywords: ANO7, AP2B1, BioID, COPG2, HSPA1A, interactome, prostate cancer, proteomics

DOI: 10.3233/CBM-190993

Citation: Cancer Biomarkers, vol. 28, no. 1, pp. 91-100, 2020

Authors: Wang, Xiaonan | Bi, Xiaomin | Huang, Xing | Wang, Bijun | Guo, Qianying | Wu, Zhengsheng

Article Type: Research Article

Abstract: BACKGROUND: ARHGDIB, a Rho GDP dissociation inhibitor protein, has been reported playing critical roles in regulation of multiple biological responses. However, whether ARHGDIB serves as a valuable biomarker in cancer is little known so far, especially in breast cancer. OBJECTIVE: In this study, we aimed to investigate the importance of ARHGDIB in breast cancer, including but not limited to biomarker-like role, as well as potential mechanisms. METHODS: Total 100 breast cancer samples and 100 benign breast disease samples were enrolled and underwent detailed pathological assessment and IHC analysis. Human breast cancer cell lines …and epithelial cell line were subjected to siRNA-mediated knock-down, RT-qPCR, western blot, MTT staining, cell cycle assay, transwell analysis respectively. RESULTS: We observed the expression of ARHGDIB is significantly higher in human breast cancer tissues compared with the benign tissues. ARHGDIB expression was positively correlated with tumor size, lymph node metastasis and TNM stage in breast cancer patients. Moreover, ARHGDIB depletion decreased proliferation, migration and invasion of breast cancer cells. Furthermore, we found ARHGDIB mediated epithelial-mesenchymal transition, and MMP2 is the key downstream effector of ARHGDIB. CONCLUSIONS: Hence, our results suggested the significance and predictive role of ARHGDIB in breast cancer. High expression of ARHGDIB indicated the poor prognosis for breast cancer patients. Show more

Keywords: ARHGDIB, biomarker, breast cancer, EMT, prognosis

DOI: 10.3233/CBM-190562

Citation: Cancer Biomarkers, vol. 28, no. 1, pp. 101-110, 2020

Authors: Guan, Gege | Wang, Yuehua | Sun, Qiushi | Wang, Ling | Xie, Fei | Yan, Jiayin | Huang, Huajun | Liu, Huijie

Article Type: Research Article

Abstract: BACKGROUND: Cancer recurrence for patients with early breast cancer is significant. Patients will benefit from more non-invasive modes of monitoring and we aim to study the feasibility of urinary circulating tumor DNA (ctDNA) to monitor for residual disease (MRD). METHODS: In this longitudinal study, 300 early breast cancer patients were recruited prospectively. Measurements were taken prior to treatment and at different time points thereafter for a total of 8 measurements. Comparisons were made with healthy volunteers and patients without detectable mutations in urine specimens. Disease free relapse were correlated to both urinary DNA quantity and ctDNA …concentration. RESULTS: Baseline index measurements showed 38% of patients with detectable mutations. The concordance with biopsy tissues was 97.3%. Overall, breast cancer patients had higher urinary DNA compared with healthy volunteers. Over time, fluctuations in urinary DNA was negligible in healthy volunteers, indicating the stability of the marker. Among the patients with detectable mutations, we observed that higher urinary DNA quantity measurements at 6-month and patients with positive mutations were associated with greater risk of relapse. Hazard ratios for patients in this category was 1.65 (95% CI 1.26–2.16) and 1.98 (95% CI 1.48–2.63) respectively. CONCLUSION: Urinary DNA offers non-invasive probing and real-time monitoring of breast cancer relapse. Our results demonstrated clear clinical relevance in breast cancer and significant risk profiling of early breast cancer patients. This potentially aids to complement current cancer relapse monitoring and may help in early intervention. Show more

Keywords: Breast cancer, biomarker, MRD, disease relapse, circulating DNA

DOI: 10.3233/CBM-190523

Citation: Cancer Biomarkers, vol. 28, no. 1, pp. 111-119, 2020

Authors: Wang, Yitao | Jiang, Feifei | Wang, Jian | Fu, Yongxing | Li, Yuanyuan | Li, Feng

Article Type: Research Article

Abstract: BACKGROUND: Non-small cell lung cancer (NSCLC) is the major type of lung cancer. MicroRNAs (miRNAs) are currently considered as novel targets and tools in cancer therapy. OBJECTIVE: The aim of this study was to investigate the expression level and functional role of miR-519a in NSCLC, as well as its clinical values. METHODS: One hundred and two patients with NSCLC were recruited. Quantitative real-time PCR (qRT-PCR) was used for the measurement of the expression level of miR-519a. Kaplan-Meier survival and Cox regression analyses were conducted to explore the prognostic significance of miR-519a in NSCLC. …MTT and Transwell assays were used to detect the effect of miR-519a on NSCLC cell proliferation, migration, and invasion. RESULTS: MiR-519a was significantly downregulated in NSCLC tissues, as well as NSCLC cell lines. The expression level of miR-519a was prominently associated with lymph node metastasis and TNM stage. Kaplan-Meier analysis suggested that low miR-519a expression was closely associated with shorter overall survival. Multivariate Cox regression analysis demonstrated that miR-519a expression level and TNM stage were two independent prognostic factors for 5-year overall survival in NSCLC patients. In vitro study, miR-519a significantly inhibited the proliferation, migration, and invasion of NSCLC cells. STAT3 was proved to be the target gene of miR-519a. CONCLUSIONS: MiR-519a functions as a tumor suppressor and inhibits tumor progression of NSCLC via targeting STAT3. MiR-519a may act as a prognostic biomarker and therapeutic target for NSCLC. Show more

Keywords: MicroRNA-519a, prognosis, progression, non-small cell lung cancer

DOI: 10.3233/CBM-190672

Citation: Cancer Biomarkers, vol. 28, no. 1, pp. 121-128, 2020