Cancer Biomarkers - Volume 22, issue 3

Authors: Chen, Bing | Li, Ya | He, Yuting | Xue, Chen | Xu, Feng

Article Type: Research Article

Abstract: Accumulating evidence suggests that long non-coding RNAs (lncRNAs) have important regulatory functions in gallbladder cancer (GBC) tumorigenesis and can serve as potential novel markers and/or targets for GBC. In this review, we critically discuss the emerging alteration of lncRNAs in GBC, the lncRNAs induced epigenetic regulation, the interaction of lncRNAs with microRNAs and lncRNAs effects on tumor-related signaling pathways. Additionally, contributions of lncRNAs in epithelial-mesenchymal transition process and energy metabolism reprogramming in GBC are also addressed. This may pave new ways towards the determination of GBC pathogenesis and lead to the development of new preventive and therapeutic strategies for GBC.

Keywords: Long non-coding RNAs, gallbladder cancer, epigenetic regulation, microRNAs, tumor-related signaling pathways

DOI: 10.3233/CBM-170979

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 359-366, 2018

Authors: Hong, Lan | Chen, Wangsheng | Wu, Dongcai | Wang, Yifeng

Article Type: Research Article

Abstract: BACKGROUND: Aberrant expression of long non-coding RNAs is involved in the progression of ovarian cancer. However, the clinical significance and biological functions of SNHG3 expression was little known in ovarian cancer (OC). METHODS: The SNHG3 expression in ovarian cancer tissues and paired adjacent normal tissues was detected using quantitative real time polymerase chain reaction (qRT-PCR). Gain-of function and loss-of function assays were performed in ovarian cancer cells to demonstrate the effects of SNHG3 expression on cell proliferation and invasion. The relative protein expression levels were determined using western blot analyses. RESULTS: The expression …of SNHG3 was significantly up-regulated in ovarian cancer tissues compared with adjacent normal tissues. Higher SNHG3 expression levels positively associated with FIGO stage, lymph node metastasis, and poor prognosis of ovarian cancer patients. Univariate and multivariate Cox regression analysis implied that FIGO stage, lymph node metastasis, higher SNHG3 expression were independent prognostic factors for overall survival (OS) rate in ovarian cancer patients. Gain-of function and loss-of function assays demonstrated that SNHG3 knockdown inhibited ovarian cancer cell proliferation and invasion abilities. However, SNHG3 overexpression promoted ovarian cancer cell proliferation and invasion abilities. Furthermore, cell proliferation and invasion related protein CyclinD1, CDK1, MMP9 and MMP3 were significantly downregulated after SNHG3 knockdown in ovarian cancer cells, while SNHG3 overexpression had reverse effects. In addition, SNHG3 functioned as an oncogene by regulating GSK3β /β -catenin signaling activity in ovarian cancer. CONCLUSIONS: Taken together, our data provide that SNHG3 has potential clinical value of and may serve as target of ovarian cancer treatment. Show more

Keywords: Ovarian cancer, long non-coding RNA, SNHG3, cell proliferation

DOI: 10.3233/CBM-170710

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 367-374, 2018

Authors: Zhao, Jingjing | Mai, Cong | Weng, Desheng | Chen, Changlong | Zhou, Ziqi | Liu, Yuan | Zhou, Zhiwei | Wang, Peng

Article Type: Research Article

Abstract: BACKGROUND: Rap1GAP, a member of the family of GTPase-activating proteins, is reported to be involved in cancer development and progression. OBJECTIVE: The study aimed to investigate the expression and prognostic value of Rap1GAP in gastric cancer patients. METHODS: Real-time quantitative polymerase chain reaction and western blotting were performed to examine Rap1GAP expression in tumorous and matched adjacent non-tumorous gastric tissues. Immunohistochemical staining was used to analyze Rap1GAP expression in 456 gastric cancer tissues. The correlation between Rap1GAP expression level and clinicopathological features as well as gastric cancer prognosis was analyzed. RESULTS: …Rap1GAP expression was remarkably decreased in tumor tissues at mRNA (p = 0.012) and protein (p = 0.034) level. Clinicopathological analysis revealed that low Rap1GAP expression was significantly correlated with tumor size (p = 0.033), histological grade (p = 0.034), T classification (p = 0.012), N classification (p = 0.006) and clinical stage (p = 0.005). Kaplan-Meier survival analysis revealed the association between low Rap1GAP expression and poor survival in gastric cancer patients. Furthermore, multivariate Cox regression analysis showed that Rap1GAP expression was an independent prognostic factor (p = 0.02). CONCLUSION: Rap1GAP may play a significant role in gastric cancer progression and act as a valuable prognostic marker for gastric cancer. Show more

Keywords: Rap1GAP, gastric cancer, prognosis, survival

DOI: 10.3233/CBM-170832

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 375-384, 2018

Authors: Wang, Dan | Wen, Gui-Min | Hou, Wei | Xia, Pu

Article Type: Research Article

Abstract: BACKGROUND: This study was carried out to investigate the correlation between CD133 and non-small cell lung cancer (NSCLC) clinicopathological features and its impact on survival of the patients with NSCLC. METHODS: In the first, we detected the level and localization of CD133 protein in NSCLC specimens and confirmed that CD133 expression was closely linked to poor prognosis of NSCLC. Secondly, TCGA genomic data for LUNG (Provisional) was retrieved and analyzed for genetic alterations of CD133 in different lung cancer subtypes. In the last, a comprehensive literature search for relevant studies published up to December 2015 was …performed using Medline, EMBASE Classic + EMBASE, and The Cochrane Central Register of Controlled Trials. RESULTS AND CONCLUSION: Both our experimental results and pooled results of previous studies indicated that CD133 may be used as a prognostic marker for the patients with NSCLC. However, more studies are required to evaluate CD133 potential use in predicting patients’ outcome. Show more

Keywords: Non-small-cell lung cancer, histology, differentiation, hazard ratio, survival

DOI: 10.3233/CBM-170835

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 385-394, 2018

Authors: Ge, Liyuan | Chen, Wei | Cao, Wenmin | Liu, Guangxiang | Zhang, Qing | Zhuang, Junlong | Zhang, Mingxin | Yang, Jun | Guo, Suhan | Zhao, Xiaozhi | Guo, Hongqian

Article Type: Research Article

Abstract: Postoperative recurrence for papillary renal cell carcinoma (PRCC) remains a tough problem in clinic. Previous studies have shown that general control nonderepressible kinase 2 (GCN2) was critically involved in tumour development. However, its function and clinical significance in renal cancer remain unknown. In this study, we investigated the role of GCN2 in PRCC. GCN2 silencing suppressed the viability and proliferation, promoted apoptosis of renal cancer cells. We found that the protein level of GCN2 was increased in PRCC tissues. Immunohistochemistry was performed in 84 patients with PRCC to explore the association of GCN2 level with clinical significance. High GCN2 protein …level was observed to be significantly correlated with adverse clinicopathological parameters, such as larger tumor size, higher TNM stage, higher Fuhurman Grade, and lymph node metastasis. We evaluated patient outcomes according to various clinical parameters as well as GCN2 expression by Kaplan-Meier curves. Multivariate analysis revealed that GCN2 overexpression can be a predictive factor correlated with reduced OS and PFS of postoperative PRCC patients. Collectively, GCN2 is potentially to play crucial roles in PRCC progression, and its overexpression may be used to predict poor prognosis and promising therapeutic strategy for PRCC patients. Show more

Keywords: GCN2, PRCC, prognosis, overall survival, progression free survival

DOI: 10.3233/CBM-170922

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 395-403, 2018

Authors: Ma, Ge | Huang, Huaxing | Li, Minghui | Li, Li | Kong, Peng | Zhu, Yichao | Xia, Tiansong | Wang, Shui

Article Type: Research Article

Abstract: Neoadjuvant chemotherapy (NCT) is the standard treatment for locally advanced breast cancer (LABC). Pathological complete response (pCR) is commonly used as a valid predictor of NCT long-term outcomes. Blood-based tumor biomarkers have the potential to predict response to NCT at early stage non-invasively. We believed plasma CCL5 could be a potential marker to predict NCT of LABC. Its efficiency and possible mechanism was studied in this work. Human Cytokine Antibody Microarray was applied to screen different cytokine concentration in plasma between low histological regression (Low-R) and high histological regression (High-R) patients. LABC patients were divided into two groups according to …pathological reactivity. The concentration of plasma CCL5 in different groups was determined by ELISA analysis. CCK8 assay was performed to analyze epirubicin susceptibility of breast cancer cells. Transwell assay was performed to determine the effect of CCL5 on breast cancer cells’ migration and invasion. qRT-PCR and western blot were used to verify the EMT (epithelial-mesenchymal transition) markers in CCL5-treated and epirubicin-treated breast cancer cells. The concentration of plasma CCL5 of Low-R group was higher than High-R group before NCT. The plasma levels of CCL5 were significantly reduced after NCT in the group of high histological regression (High-R). Epirubicin susceptibility decreased in the breast cancer cells treated by recombinant CCL5. Migration and invasion were significantly enhanced in breast cancer cells treated by recombinant CCL5. E-cadherin expression was decreased whereas vimentin increased significantly in CCL5-treated breast cancer cells. The phosphorylation of ezrin in Y-567 and its downstream protein cortactin increased significantly in CCL5-treated breast cancer cells. Plasma CCL5 level could be a promised candidate to predict chemotherapy response of breast cancer. Plasma CCL5 plays an important role in EMT process of breast cancer. Show more

Keywords: Neoadjuvant chemotherapy, breast cancer, CCL5, predictive marker

DOI: 10.3233/CBM-170986

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 405-415, 2018

Authors: Liu, De-Gang | Xue, Lei | Li, Jun | Yang, Qiang | Peng, Jiang-Zhou

Article Type: Research Article

Abstract: BACKGROUND: Circulating tumor cells (CTCs) is a promising biomarker for cancer prognosis and monitoring. Molecular characterizing of CTCs could provide beneficial information on the basis of CTCs counting. OBJECTIVE: To investigate the epithelial-mesenchymal transition (EMT) phenotypes and GALC mRNA expression of CTCs in non-small cell lung cancer (NSCLC) patients. METHODS: We analyzed the baseline number, EMT classification, and GALC expression of CTCs in 47 NSCLC patients using CanPatrol platform and RNA in situ hybridization technique. RESULTS: CTCs were detected in 91.5% patients ranging 0–47/5 mL blood. Increased CTCs were …associated with advanced tumor stages (6/5 mL) compared with early stages (3.5/5 mL). Patients with effective treatment response presented lower CTCs (3.5/5 mL) than patients with insufficient response (7/5 mL). Epithelial, hybrid and mesenchymal CTCs were detected in 55.4%, 78.7% and 61.7% patients, respectively. Patients with distant metastasis and poor curative outcomes presented higher level of EMT-CTCs. GALC expression was positive in CTCs of 80.6% patients and closely correlated with tumor number and distant metastasis and treatment outcomes. CONCLUSIONS: EMT phenotypes and GALC expression of CTCs are correlated with cancer metastasis and therapeutic outcomes, suggesting them to be potential markers for the prognosis of NSCLC. Show more

Keywords: CTCs detection, cancer metastasis, circulating biomarkers, therapeutic monitoring and prognosis

DOI: 10.3233/CBM-170995

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 417-426, 2018

Authors: Ding, Wei | Tan, Hongbo | Li, Xuemei | Zhang, Yue | Fang, Fang | Tian, Yuanyuan | Li, Jin | Pan, Xinghua

Article Type: Research Article

Abstract: BACKGROUNDS: MicroRNAs (miRNAs) are some RNA molecules that negatively regulate gene expression by binding to the 3’-untranslated region (3’-UTR) of target mRNA molecules. The aim of the study is to investigate the clinical role and functional effects of microRNA-493 (miR-493) in human hepatocellular carcinoma (HCC). METHODS: Expression of miR-493 in 58 cases of HCC tissues and adjacent normal tissues was determined by using quantitative real-time PCR (qRT-PCR) analyses. In vitro , cell proliferation and invasion capacity was evaluated by CCK8 assay and trans-well invasion assay. Luciferase reporter assay, western blot and qRT-PCR were used to detect …the association between miR-493 expression and zinc finger protein X-linked (ZFX) in HCC. RESULTS: Expression of miR-493 was significantly downregulated in HCC tissues compared to adjacent normal tissues. Lower miR-493 expression associated with tumor size, vascular invasion and poor overall survival (OS) and disease free survival (DFS) time of HCC patients. In vitro , transfection of miR-493 mimic in HCC cells inhibited cell proliferation and invasion abilities. However, transfection of miR-493 inhibitor in HCC cells had promoting effects. Luciferase reporter assay, qRT-PCR and western blot analysis demonstrated that miR-493 targeting 3’-untranslated region (3’-UTR) of ZFX and overexpression of miR-493 inhibited its expression. Moreover, enforced ZFX expression rescued the effects of miR-493 mimic on cell proliferation and invasion. CONCLUSION: MiR-493 functioned as tumor suppressor in HCC cells by regulating ZFX expression. Thus, miR-493 may provide potential value for HCC treatment. Show more

Keywords: MicroRNAs, miR-493, ZFX, cell proliferation, cell invasion

DOI: 10.3233/CBM-171036

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 427-434, 2018

Authors: Wang, Ning | Che, Yanci | Yin, Fufen | Yu, Fengsheng | Bi, Xiaoning | Wang, Yankui

Article Type: Research Article

Abstract: BACKGROUND: Cervical cancer is one of the malignant tumors which seriously threaten the women health worldwide. SPINT2 is an endogenous inhibitor of hepatocyte growth factor activator and down regulated or even silenced in many human malignant tumors. OBJECTIVE: This study was performed to explore the promoter methylation status of SPINT2 gene and the effect on its expression in cervical carcinoma. METHODS: HPV-positive and -negative cervical cancer cell lines, 50 cases of cervical carcinoma tissues, and 20 cases of normal cervical tissues were used for this study. The methylation status of promoter and the …first exon of SPINT2 gene were analyzed. The expression of SPINT2 was analyzed by qRT-PCR. RESULTS: HPV E6/E7 infection affects SPINT2 methylation rate in cell lines. SPINT2 methylation rate of HT-3E6/E7 was 8.8%, while the methylation rate of SPINT2 in HT-3 was 0%. In cervical tissues, the methylation rate of SPINT2 in cervical cancers was 54%, while the methylation rate of SPINT2 in normal cervical samples was 25%. As for cervical cancers, the methylation rate of SPINT2 gene was higher in grade 3 than those of grade 2. CONCLUSIONS: The expression of SPINT2 gene is regulated by its methylation status, and the methylation status of SPINT2 is altered by HPV infection. The aberrant methylation status of SPINT2 gene may play an important role in the development of cervical cancer. Show more

Keywords: SPINT2, cervical carcinoma, HPV, methylation, 5-Aza-CdR

DOI: 10.3233/CBM-171050

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 435-442, 2018

Authors: Ma, Shuyun | Rong, Xuan | Gao, Fei | Yang, Yang | Wei, Lin

Article Type: Research Article

Abstract: BACKGROUND: Targeting protein for Xenopus kinesin-like protein 2 (TPX2) is a microtubule-associated proteinrequired for mitosis and spindle assembly. It has been revealed that TPX2 is overexpressedin various human cancers and promotes cancer progression. METHODS: The expression of TPX2 was examined in ovarian cancer (OC) tissues and by Western blotting, quantitative real-time reverse transcription PCR (qRT-PCR) and immunohistochemistry. The effects of TPX2 on proliferation and migration of two OC cell lines SKOV3and RMG1 were analyzed using the methylthiazol tetrazolium (MTT) assay, flow cytometry and transwell assay. The mechanisms underlying the effects of TPX2 on OC cells were …explored by qRT-PCR and Western blot. RESULTS: In this study, we found that TPX2 was upregulated in OC tissues. We observed knockdown of TPX2 inhibited the expression of Polo-like kinase 1 (PLK1), which has an important role in the regulation of M phase of the cell cycle, and the activity of Cdc2, induced cell arrested at the G2/M phase and decreased proliferation. Moreover, our data revealed that the levels of PLK1, β -catenin, MMP7 and MMP9 were inhibited following TPX2 knockdown, leading to decrease of cell migration. Finally, we showed that the restoration of PLK1 expression attenuated the anti-proliferation and anti-migration effects of TPX2 knockdown in OC cells. CONCLUSIONS: TPX2 promotes the proliferation and migration of human OC cells by regulating PLK1 expression. Show more

Keywords: Ovarian cancer, TPX2, OC cell line, proliferation, migration

DOI: 10.3233/CBM-171056

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 443-451, 2018

Authors: Liu, Zhuo | Ma, Min | Yan, Lei | Chen, Shilin | Li, Sha | Yang, Darong | Wang, Xiaohong | Xiao, Hua | Deng, Hongyu | Zhu, Haizhen | Zuo, Chaohui | Xia, Man

Article Type: Research Article

Abstract: BACKGROUND: Interferon-α (IFN-α ) is an adjuvant to chemotherapy and radiotherapy for hepatocellular carcinoma (HCC), but some HCC patients do not respond to treatment with IFN-α . METHODS: We performed loss-of-function and gain-of-function experiments to examine the role of ISG15 in the IFN-α sensitivity of LH86, HLCZ01, SMMC7721, and Huh7 cell lines and tumor samples. RESULTS: The overexpression of ISG15 reduced apoptosis in Huh7 and LH86 cells in the presence of IFN-α , whereas the shRNA-mediated knock down of ISG15 expression …increased apoptosis in both Huh7 and LH86 cells. We identified a putative miR-370 target site in the 3’-UTR in the ISG15 mRNA, and the level of miR-370 expression in HCC cell lines reflected the level of IFN-α -induced apoptosis exhibited by each. Both HCC cell lines and tumor samples had significantly lower levels of miR-370 than the control cells and tissues (P < 0.05). The overexpression of miR-370 in IFN-α -treated LH86 and Huh7 cells increased apoptosis and reduced the volume of LH86- and Huh7-derived xenograft tumors in mice treated with IFN-α compared with the control tumors. CONCLUSIONS: Our findings suggest that miR-370 functions as an HCC tumor suppressor and regulator of IFN-α sensitivity and that miR-370 might be a useful prognostic marker for HCC patients. Show more

Keywords: Hepatocellular carcinoma cells, interferon-α, ISG15, miR-370, prognostic marker

DOI: 10.3233/CBM-171075

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 453-466, 2018

Authors: Zhuo, Minglei | Chen, Hanxiao | Zhang, Tianzhuo | Yang, Xue | Zhong, Jia | Wang, Yuyan | An, Tongtong | Wu, Meina | Wang, Ziping | Huang, Jing | Zhao, Jun

Article Type: Research Article

Abstract: BACKGROUND: The PD-L1 antibody atezolizumab has shown promising efficacy in patients with advanced non-small cell lung cancer. But the predictive marker of clinical benefit has not been identified. OBJECTIVE: This study aimed to search for potential predictive factors in circulating blood of patients receiving atezolizumab. METHODS: Ten patients diagnosed with advanced non-small cell lung cancer were enrolled in this open-label observing study. Circulating immune cells and plasma tumor markers were examined in peripheral blood from these patients before and after atezolizumab treatment respectively. Relation between changes in circulating factors and anti-tumor efficacy were …analyzed. RESULTS: Blood routine test showed that atezolizumab therapy induced slightly elevation of white blood cells count generally. The lymphocyte ratio was increased slightly in disease controlled patients but decreased prominently in disease progressed patients in response to atezolizumab therapy. Flow cytometric analysis revealed changes in percentage of various immune cell types, including CD4 + T cell, CD8 + T cell, myeloid-derived suppressor cell, regulatory T cell and PD-1 expressing T cell after atezolizumab. Levels of plasma tumor marker CEA, CA125 and CA199 were also altered after anti-PD-L1 therapy. In comparison with baseline, the disease progressed patients showed sharp increase in tumor marker levels, while those disease controlled patients were seen with decreased regulatory T cell and myeloid-derived suppressor cell ratios. CONCLUSIONS: The circulating immune cell ratios and plasma tumor marker levels were related with clinical efficacy of atezolizumab therapy. These factors could be potential predictive marker for anti-PD-L1 therapy in advanced non-small cell lung cancer. Show more

Keywords: Circulating immune cell, immunotherapy, non-small cell lung cancer, PD-L1, tumor marker

DOI: 10.3233/CBM-171089

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 467-476, 2018

Authors: Huang, Qing-Xing | Ma, Jun | Wang, Yu-Sheng

Article Type: Research Article

Abstract: BACKGROUND: Oxidative stress plays an important role in promoting proliferation and metastases of cancer, which can be represented by ischemia-modified albumin (IMA). The purpose of this study was to evaluate serum IMA level in patients with operable advanced gastric cancer and analyze its prognostic significance. MATERIALS AND METHODS: A total of 274 patients with primary stage III gastric cancer underwent curative operation were enrolled in this study. Serum IMA level was measured within 24 hours before surgery, comparing with 112 healthy donors. The correlation between serum IMA level and survival outcome was analyzed by the Kaplan-Meier …with Log-Rank test and Cox’s regression methods, respectively. RESULTS: Serum IMA level from gastric cancer was higher than healthy control (0.41 ± 0.12 VS 0.23 ± 0.08; P < 0.001). Finally, 173 and 181 patients out of all 274 patients studied had died and recurrent, respectively. All patients were stratified into two groups using the optimal cutoff value (0.45) of IMA level using a sensitivity of 92.5% and a specificity of 65.2% as optimal conditions from receiver operating curve analysis. Patients with a IMA ⩾ 0.45 had poorer mean overall survival (44.68 months VS 30.94 months, P = 0.010) and mean recurrence free survival (42.36 months VS 28.82 months, p = 0.01) than patients with a IMA < 0.45 in univariate analysis and IMA also been confirmed as independent predictor for survival for GC patients in multivariate analysis (OR, 0.731; 95% CI: 0.329–1.282; p = 0.023). CONCLUSIONS: Serum IMA level can be considered as an independent prognostic factor for operable and advanced gastric cancer. Show more

Keywords: Ischemia-modified albumin, survival, gastric cancer

DOI: 10.3233/CBM-171090

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 477-485, 2018

Authors: Liu, Wenjun | Xu, Yongmei | Guan, Hongliang | Meng, Hongwei

Article Type: Research Article

Abstract: BACKGROUND: Breast cancer remains the most invasive female malignancy worldwide. Functional role of microRNA-940 (miR-940) have been investigated in various cancer. The purpose of this study was to assess the serum miR-940 expression and its clinical significance in breast cancer. METHODS: Expression of miR-940 was measured by quantitative real-time polymerase chain reaction (qRT-PCR). The diagnostic value of miR-940 was analyzed with receiver operating characteristics (ROC) analysis. To explore the prognostic performance of miR-940, Kaplan-Meier survival assay and Cox regression analysis were performed. RESULTS: Downregulated miR-940 was detected in the breast cancer patients compared …with the healthy controls (P < 0.001). The miR-940 expression was correlated with lymph node metastasis (P = 0.014) and TNM stage (P = 0.003). The area under the ROC curve (AUC) was 0.905, with sensitivity and specificity of 94.5% and 78.6%. From the survival curves, patients with low miR-940 expression had poor overall survival compare with those with high expression (log-rank P = 0.009). The Cox analysis indicated that miR-940 was an independent prognostic factor (HR = 2.645, 95% CI = 1.426–4.906 and P = 0.002). Decreased miR-940 expression was also been found in triple-negative breast cancer (TNBC) samples, and might predict poor prognosis in TNBC patients. CONCLUSIONS: Serum downregulated miR-940 may serve as a reliable diagnostic and prognostic biomarker in breast cancer patients. Show more

Keywords: MiR-940, diagnosis, prognosis, breast cancer

DOI: 10.3233/CBM-171124

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 487-493, 2018

Authors: Li, Yanshi | Liu, Chuan | Wang, Zhihai | Hu, Guohua

Article Type: Research Article

Abstract: Protocadherin8 (PCDH8), an integral membrane protein, was reported to be a tumor suppressor involved in tumorigenesis in cancers. We aimed to investigate the expression of PCDH8 and its clinicopathological significance in hypopharyngeal carcinoma. We also examined the possible inactivation mechanism of PCDH8. A total of 80 pairs of hypopharyngeal carcinoma tumor tissues and non-tumor tissues were investigated to examine the immunohistochemical expression of PCDH8. Prognostic value and clinicopathological significance of PCDH8 expression were examined by Kaplan-Meier and log-rank test and Cox’s ones. Ten pairs of tumor tissues and non-tumor tissues were analyzed by RT-PCR and 4 pairs by Western blot …respectively. Promoter methylation of PCDH8 was observed in 14 pairs of tumor tissues and non-tumor tissues by methylation-specific PCR (MSP). The expression of PCDH8 in tumor tissues was depressed immunohistochemically when compared with non-tumor tissues and was significantly lower in the advanced pathological stage. Meanwhile, the expression of PCDH8 served as an independent prognostic risk factor for overall survival of hypopharyngeal carcinoma. The mRNA and protein levels of PCDH8 in tumor tissues was also down-regulated than in non-tumor tissues. Moreover, aberrant promoter methylation of PCDH8 occurred frequently in tumor tissues, rather than in non-tumor tissues. For the first time, our study demonstrates the tumor-suppressive function of PCDH8 and its epigenetic inactivation by promoter methylation in hypopharyngeal carcinoma. It suggests that PCDH8 may serve as a useful prognostic biomarker and a potential therapeutic target for hypopharyngeal carcinoma patients. Show more

Keywords: Hypopharyngeal carcinoma, protocadherin8, tumor suppressor gene

DOI: 10.3233/CBM-171137

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 495-502, 2018

Authors: Mekky, Mohamed A. | Salama, Rgaa H. | Abdel-Aal, Mahmoud F. | Ghaliony, Mohamed A. | Zaky, Saad

Article Type: Research Article

Abstract: BACKGROUND: Data about the molecular pathogenesis of hepatitis C-related hepatocellular carcinoma (HCC) are still challenging. OBJECTIVES: Therefore, we tried to investigate the epigenetic study of three nominated genes (APC, P14, and E-cadherin) in the pathogenesis of HCV-related HCC in Egyptian. METHODS: Between March 2016 and March 2017, the DNA methylation, and quantification using (epigenetic ELISA kit) for E-cadherin, APC, and P14 genes were studied in three groups of patients: HCV related liver cirrhosis without HCC group (LC-group; n = 20), HCC on top of HCV-related cirrhosis (HCC-group; n …= 20), and a third apparently healthy control group (control-group; n = 10). RESULTS: E-cad methylation showed non-significant differences between groups. P14 methylation was occurred only in HCC-group (45%). APC methylation was the highest in HCC group (70%). Methylation level was high in HCC group in comparison to both LC and control groups (P < 0.001). DNA methylation at a cutoff point > 2.9 ng/ml predicts HCC in LC-group with 90% sensitivity and 80% specificity and at level > 2.3 ng/ml had 95% sensitivity and 90% specificity in control-group. The pooled sensitivity, specificity, positive and negative predictive values and accuracy were 90%, 60%, 69.2, 85.7 and 75% respectively. CONCLUSION: Aberrant DNA methylation of multiple genes is associated with disease progression in HCV related cirrhosis. Moreover, early detection of promotor methylation of these may sever as good biomarker for early detection and therapeutic targets in high risk patients. Show more

Keywords: Hepatocellular carcinoma, liver cirrhosis, epigenetics

DOI: 10.3233/CBM-171156

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 503-509, 2018

Authors: Zhang, Liang | Huang, Yi | Ling, Junjun | Zhuo, Wenlei | Yu, Zhen | Shao, Mengmeng | Luo, Yunbo | Zhu, Yi

Article Type: Research Article

Abstract: BACKGROUND: Liver carcinoma is a major cause of cancer-related death worldwide. Up to date, the mechanisms of liver cancerigenesis and development have not been fully understood. Multi-genes and pathways were involved in the tumorigenesis of liver cancer. OBJECTIVE: The aim of the present study was to screen key genes and pathways in liver cancerigenesis and development by using bioinformatics methods. METHODS: A dataset GSE64041 were retrieved from GEO database and the differentially expressed genes (DEGs) were screened out. Then the DEG functions were annotated by gene ontology (GO) and pathway enrichment analysis, respectively. …The hub genes were further selected by protein-protein interaction (PPI) analysis. Afterwards, the mRNA and protein expressions as well as the prognostic values of the hub genes were assessed. RESULTS: As a result, 208 up-regulated and 82 down-regulated genes were screened out. These DEGs were mainly enriched in cell cycle and metabolism-related pathways. Through PPI analysis, TOP2A, PRDM10, CDK1, AURKA, BUB1, PLK1, CDKN3, NCAPG, BUB1B and CCNA2 were selected as hub genes, which were all over-expressed in liver cancers relative to those in normal tissues, respectively. Among them, PLK1 and CCNA2 were suggested to be prognostic factors for liver carcinoma. CONCLUSION: In conclusion, the present study identified several hub genes, and cell cycle and metabolism-related pathways that may play critical roles in the tumorigenesis of liver cancer. Future validation laboratory experiments are required to confirm the results. Show more

Keywords: Liver carcinoma, hub genes, cell cycle, prognosis, bioinformatics

DOI: 10.3233/CBM-171160

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 511-521, 2018

Authors: Ji, Yu | Shao, Zhenzhen | Liu, Junjun | Hao, Yujuan | Liu, Peifang

Article Type: Research Article

Abstract: This study aimed to analyze the correlation between mammographic density obtained by density analysis software (DAS)/radiologists visual (RV) classification with molecular subtype, and the expression levels of estrogen receptor (ER), progesterone receptor (PR), Ki67 antigen (Ki-67), p53 gene (p53), and human epidermal growth factor receptor-2 (HER2). A total of 688 breast cancer patients with digital mammography and complete molecular pathological results in Tianjin Medical University Cancer Institute and Hospital between February 2015 and February 2016 were collected. The DAS-density grade (DASD) and the radiologists visually classified density grade (RVD) were evaluated by 3 radiologists. The correlation between density grade and …the expression levels of ER, PR, Ki-67, p53, HER2 and breast cancer molecular subtype (PMS) were analyzed. The agreement between DASD and RVD was explored. ER, PR and HER-2 positive rate were significantly different among patients with different RVD grades (P < 0.05). HER2 positive rates showed an increasing trend following RVD upgrading (P 𝑡𝑟𝑒𝑛𝑑 < 0.05). HER-2 positive rate in RVD D1 + D2 was 7.69%, which was higher than that in D3 + D4 (P < 0.05). The ER and Ki-67 expressions in patients were markedly different among DASD (P = 0.009 and 0.002) and RVD (P = 0.012 and 0.036) with different grades. The kappa value of each DASD to RVD was 0.31 (P < 0.01). The RVD 3 proportion was 14.58% (63/432) in HER2 Over-expressing subtype, which was apparently higher than RVD1 (2.43%, 1/41) (P < 0.05). Breast density may be partial correlated with molecular pathology in breast cancer. Show more

Keywords: Breast cancer, mammographic density, molecular pathology, correlation

DOI: 10.3233/CBM-181185

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 523-531, 2018

Authors: Wang, Qiang | Peng, Ruixian | Wang, Boshi | Wang, Jifeng | Yu, Wandong | Liu, Yongzhong | Shi, Guowei

Article Type: Research Article

Abstract: BACKGROUND: Krüppel-like factor 13 (KLF13), a member of the KLF family, is involved in the development of immunological diseases and tumor progression. However, the expression patterns and potential functions of KLF13 in prostate carcinoma are still unknown. Here, we aimed to study the roles and mechanisms of KLF13 in prostate cancer. METHODS: The expression levels of KLF13 was detected by Immunohistochemistry in prostate tumor tissues and the paired non-tumor tissues. The effects of KLF13 up-regulation was tested by performing CCK8, cell colon formation, flow cytometric analysis and measurement of tumor proliferation in nude mice. Signaling pathway …was analyzed by Western blot. RESULTS: The current study, for the first time, found that KLF13 was downregulated in prostate tumor tissues as compared to the paired non-tumor tissues. The overexpression of KLF13 dramatically inhibited cell proliferation and induced apoptosis by suppressing the AKT pathway in human prostate cancer cells. Moreover, the ectopic expression of KLF13 efficiently delayed the onset of PC3 xenografts and inhibited the tumor growth in vivo. CONCLUSIONS: KLF13 functions as a tumor suppressor protein in PCa, and the pharmacological activation of KLF13 might represent a potential approach for the treatment of prostate cancer. Show more

Keywords: Immunohistochemistry, prostate cancer, apoptosis, AKT pathway, Krüppel-like factor 13

DOI: 10.3233/CBM-181196

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 533-541, 2018

Authors: Jiang, Sixiong | Tian, Feng | Wang, Qi | Cheng, Wen | Wang, Longxin | Wang, Ying | Sun, Weibing

Article Type: Research Article

Abstract: BACKGROUND: Spindle and kinetochore-associated protein 1 (SKA1) is a component of SKA, which is essential for proper chromosome segregation. Recently, SKA1 was found to be over-expressed in several types of human cancers. However, reports on the relationship between SKA1 expression and the prognosis of bladder cancer, in particular, are lacking. OBJECTIVES: To clarify the clinical significance of SKA1 as a prognostic biomarker for early recurrence and progression of patients with non-muscle invasive bladder cancer (NMIBC). METHODS: The differential expression levels of SKA1 of 148 NMIBC tissues were determined by immunohistochemical staining. Quantitative real-time …PCR and western blot analysis were further performed to confirm the immunohistochemistry results. Recurrence and progression free interval were assessed by Kaplan-Meier method and differences between groups calculated by log-rank statistics. The prognostic value of SKA1 for early recurrence and progression was analyzed by multivariate Cox proportional hazard regression models. RESULTS: SKA1 expression was significantly different in various NMIBC tissues. Kaplan-Meier analysis revealed that patients with high SKA1 expression showed high early recurrence (p < 0.001) and progression (p < 0.05) rates. Although univariate Cox regression analysis revealed that several other factors had an impact on recurrence and progression, upon multivariate analysis, high SKA1expression was the only independent predictor for early recurrence (hazards ratio [HR], 0.246; 95% confidence interval [CI], 0.131–0.461; p = 0.000) and progression (HR, 0.194; 95% CI, 0.052–0.715; p = 0.014). CONCLUSIONS: High SKA1 expression is associated with early recurrence and progression in patients with NMIBC, indicating SKA1 may serve as a promising prognostic biomarker for this disease. Show more

Keywords: Non-muscle invasive bladder cancer, SKA1, recurrence, progression, biomarker

DOI: 10.3233/CBM-181202

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 543-549, 2018

Authors: Tao, Zhihang | Li, Stanley Xiangyu | Cui, Xiwei | Huang, Yamin | Zhu, Sha | Wang, Yexiao | Tan, Huixin | Ma, Xuelei

Article Type: Research Article

Abstract: BACKGROUND: Neutrophil-Lymphocyte Ratio (NLR) and Platelet-Lymphocyte Ratio (PLR) have been considered as indicators for prognosis in various cancers. However, the prognostic values of NLR and PLR have never been tested in gallbladder carcinoma (GBC) with hepatic involvement. OBJECTIVE: The aim of the current study was to assess the prognostic significance of NLR, PLR, and other candidate biomarkers in GBC with liver involvement. METHODS: Receiver operating characteristic (ROC) curve analyses were utilized to pinpoint the cut-off values for NLR, PLR, and Monocyte-Lymphocyte Ratio (MLR). Univariate analyses were employed to estimate the impact of NLR, …PLR, MLR, and other inflammatory indexes on median survival. Multivariate analyses were used to verify the independent prognostic predictors. RESULTS: Eighty four patients were enrolled from 2009 to 2017. The cut-off values for NLR, PLR, and MLR were 3.20, 117.75, and 0.25, respectively. Univariate analyses revealed that TNM stage, NLR, PLR, MLR, lactate dehydrogenase, alkaline phosphatase, and carcinoembryonic antigen were significantly associated with decreased survival in GBC with hepatic involvement. Advanced TNM stage (P < 0.001) and elevated preoperative NLR (P = 0.002) were significantly associated with lower median survival periods, as revealed by multivariate analyses. CONCLUSIONS: These findings suggest that preoperative NLR may be an independent prognostic factor in evaluating prognosis in GBC with liver involvement. Show more

Keywords: Gallbladder cancer, immunologic markers, overall survival, prognostic factors, serum markers

DOI: 10.3233/CBM-181230

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 551-557, 2018

Authors: Shen, Wenjie | Cui, Ming-Ming | Wang, Xin | Wang, Rui-Tao

Article Type: Research Article

Abstract: BACKGROUND: Esophageal cancer (EC) is the sixth most common cause of death from cancer. Altered mean platelet volume (MPV) levels were found in patients with malignancies. OBJECTIVE: The present study investigated whether MPV can predict the survival in EC patients. MEHTODS: The clinical data of 236 consecutive EC patients between January 2009 and December 2009 in our center were retrospectively analyzed. The overall survival rate was estimated using Kaplan-Meier method. Cox proportional hazards models were fitted to model the relationships between patient characteristics and prognosis. RESULTS: Decreased MPV was significantly …correlated with tumor location and tumor differentiation (p < 0.001). Moreover, survival analysis revealed that the overall survival of patients with MPV ⩽ 7.4 fL was significantly shorter than that of those with MPV > 7.4 fL. Multivariate analysis identified MPV as an independent poor prognostic factor for overall survival. CONCLUSIONS: Reduced MPV is associated with worse survival outcome in EC. Show more

Keywords: Esophageal cancer, mean platelet volume, prognosis

DOI: 10.3233/CBM-181231

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 559-563, 2018

Authors: Song, Rui | Zhang, Jia | Huang, Junhua | Hai, Tao

Article Type: Research Article

Abstract: OBJECTIVE: Breast cancer is a common malignancy in women and long non-coding RNAs (lncRNAs) have been shown to play key roles in the development and progression of breast cancer. In the present study, we examined the biological role of lncRNA gastric carcinoma highly expressed transcript 1 (GHET1) in breast cancer. METHODS: The expression of GHET1 was determined by qRT-PCR assay; CCK-8, colony formation, Transwell invasion and migration assays detected breast cancer cell proliferation, invasion and migration; cell apoptosis and cell cycle were determined by flow cytometry; protein levels were determined by western blot assay. …RESULTS: GHET1 was up-regulated in breast cancer tissues and cell lines, and the up-regulation of GHET1 was positively correlated with larger tumor size, advanced clinical stage, lymph node metastasis and shorter overall survival. Knockdown of GHET1 suppressed cell proliferation, invasion and migration, and induced apoptosis and G 0 /G 1 cell cycle arrest in MCF-cells. Knockdown of GHET1 also suppressed the protein levels of N-cadherin, vimentin, and decreased the protein level of E-cadherin in MCF-7 cells. On the other hand, overexpression of GHET1 promoted cell proliferation, invasion and migration, and inhibited cell apoptosis and increased cell population at S phase in BT-20 cells. Overexpression of GHET1 also promoted epithelial mesenchymal transition (EMT) in BT-20 cells. Furthermore, knockdown of GHET1 also suppressed in vivo tumor growth of MCF-7 cells, and also decreased the protein levels of N-cadherin and vimentin, and increased the protein levels of E-cadherin in the tumor tissues from the nude mice. CONCLUSIONS: Our results demonstrated that GHET1 was up-regulated in breast cancer tissues and cell lines, and promoted breast cancer cell proliferation, invasion and migration by affecting EMT. Our study for the first time revealed the biological functions of GHET1 in breast cancer. Show more

Keywords: Breast cancer, GHET1, cell proliferation, invasion and migration, apoptosis, epithelial mesenchymal transition

DOI: 10.3233/CBM-181250

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 565-573, 2018

Authors: Ma, De-Qiang | Zhang, Yin-Hua | Ding, De-Ping | Li, Juan | Chen, Lin-Li | Tian, You-You | Ao, Kang-Jian

Article Type: Research Article

Abstract: OBJECTIVE: To investigate the impact of Bmi-1-mediated NF-κ B pathway on the biological characteristics of CD133 + liver cancer stem cells (LCSCs). METHODS: Flow cytometry was used to isolate CD133 + LCSC cells from Huh7, Hep3B, SK-hep1, and PLC/PRF-5 cells. CD133 + Huh7 cells were divided into Control, Blank, Bmi-1 siRNA, JSH-23 (NF-κ B pathway inhibitor), and Bmi-1 + JSH-23 groups. The properties of CD133 + Huh7 cells were detected by the colony-formation …and sphere-forming assays. Besides, Transwell assay was applied for the measurement of cell invasion and migration, immunofluorescence staining for the detection of NF-κ B p65 nuclear translocation, and qRT-PCR and Western blotting for the determination of SOX2, NANOG, OCT4, Bmi-1, and NF-κ B p65 expression. RESULTS: CD133 + Huh-7 cells were chosen as the experiment subjects after flow cytometry. Compared with CD133 - Huh-7 cells, the expression of CD133, OCT4, SOX2, NANOG, Bmi-1, and NF-κ B p65, the nuclear translocation of NF-κ B p65, the number of cell colonies and Sphere formation, as well as the abilities of invasion and migration were observed to be increased in CD133 + Huh-7 cells, which was inhibited after treated with Bmi-1 siRNA or JSH-23, meanwhile, the cell cycle was arrested at the G0/G1 and S phases with apparently enhanced cell apoptosis. Importantly, no significant differences in the biological characteristics of CD133 + Huh-7 cells were found between the Blank group and Bmi-1 + JSH-23 group. CONCLUSION: Down-regulating Bmi-1 may inhibit the biological properties of CD133 + LCSC by blocking NF-κ B signaling pathway, which lays a scientific foundation for the clinical treatment of liver cancer. Show more

Keywords: Liver cancer stem cells, Bmi-1, NF-κB pathway, CD133+

DOI: 10.3233/CBM-181329

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 575-585, 2018

Authors: Zahran, Asmaa M. | Mohammed Saleh, Mostafa F. | Sayed, Mona M. | Rayan, Amal | Ali, Arwa Mohammed | Hetta, Helal F.

Article Type: Research Article

Abstract: BACKGROUND: The bone marrow immunosuppressive microenvironment of AML patients sustains and modulates proliferation, survival and drug resistance of AML through deregulation of both innate and adaptive immune response. We aimed to investigate the level of Tregs, expression of Tim-3 on peripheral blood T cells, expression of CD200 in myeloid blasts in newly diagnosed AML patients with normal cytogenetics (AML-NC) and their prognostic impact. PATIENTS AND METHODS: This study included 40 patients with de novo AML-NC and 20 healthy controls. Flow-cytometry was used for detection of CD4+ CD25+ high FoxP3+ regulatory T …cells, Tim-3 expression on peripheral blood T cells and CD200 expression on myeloid blasts. RESULTS: The percentages of CD4+ CD25+ high and CD4+ CD25+ high Foxp3+ Tregs were significantly increased in AML patients than controls. The levels of Tregs, Tim-3/CD4 + , Tim-3/CD8 + , CD200 and MFI of CD200 were significantly lower in responding patients than in those with persistent leukemia. Only high CD200 expression (> 50%) showed statistically significant worse OS with P < 0.04. CONCLUSION: The increased levels of Tregs, Tim-3 expression on peripheral blood T cells and CD200 expression in myeloid blast in AML patients could play a role in the development of AML. Analysis of these markers could serve as prognostic markers and might guide the therapy in AML patients in the future. Show more

Keywords: Regulatory T cells, CD200, TIM3, acute myeloid leukemia, normal cytogenetics

DOI: 10.3233/CBM-181368

Citation: Cancer Biomarkers, vol. 22, no. 3, pp. 587-595, 2018