Cancer Biomarkers - Volume 29, issue 2

Authors: Pan, Xue | Chen, Ying | Gao, Song

Article Type: Research Article

Abstract: BACKGROUND: Ovarian cancer is the common tumor in female, the prognostic of which is influenced by a series of factors. In this study, 4 genes relevant to pathological grade in ovarian cancer were screened out by the construction of weighted gene co-expression network analysis. METHODS: GSE9891 with 298 ovarian cancer cases had been used to construct co-expression networks. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses was used to analyze the possible mechanism of genes involved in the malignant process of ovarian cancer. Hub genes were validated in other independent datasets, such as …GSE63885, GSE26193 and GSE30161. Survival analysis based on the hub genes was performed by website of Kaplan Meier-plotter. RESULTS: The result based on weighted gene co-expression network analysis indicated that turquoise module has the highest association with pathological grade. Gene Ontology enrichment analysis revealed that the genes in turquoise module main enrichment in inflammatory response and immune response. Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that the genes in turquoise module main enrichment in cytokine-cytokine receptor interaction and chemokine signaling pathway. In turquoise module, a total of 4 hub genes (MS4A4A, CD163, CPR65, MS4A6A) were identified. Then, 4 hub genes were effectively verified in the test datasets (GSE63885, GSE26193 and GSE30161) and tissue samples from Shengjing Hospital of China Medical University. Survival analysis indicated that the 4 hub genes were associated with poor progression-free survival of ovarian cancer. CONCLUSIONS: In conclusion, 4 hub genes (MS4A4A, CD163, CPR65, MS4A6A) were verified associated with pathological grade of ovarian cancer. Moreover, MS4A4A, CD163, MS4A6A may serve as a surface marker for M2 macrophages. Targeting the 4 hub genes may can improve the prognosis of ovarian cancer. Show more

Keywords: Ovarian cancer, WGCNA, pathological grade, M2 macrophages, prognosis

DOI: 10.3233/CBM-191162

Citation: Cancer Biomarkers, vol. 29, no. 2, pp. 169-178, 2020

Authors: Xuan, Yun-Ze | Jin, Cheng-Ri | Yang, Kang-Juan

Article Type: Research Article

Abstract: OBJECTIVE: The aim of this study was to explore the mechanisms by which oral cancer acquires resistance to gemcitabine. METHODS: Oral squamous cell carcinoma (OSCC) cells were treated with gemcitabine upon infection or with a lentivirus harboring short hairpin RNA (shRNA) targeted to transforming growth factor-β (TGF-β ). Then, Western blot, ELISA, migration assay, MTT assay, and animal experiments were used to explore the mechanism of resistance to gemcitabine treatment. RESULTS: After the treatment of non-transfected cells with gemcitabine, NF-κ B and AKT activities were increased, …which may have induced the OSCC resistance to gemcitabine. Then, we found that TGF-β downregulation effectively reduced NF-κ B and AKT phosphorylation levels after the administration of gemcitabine and increased the OSCC sensitivity to gemcitabine, resulting in cell death and the blunting of OSCC resistance to gemcitabine. The EMT was also reduced by TGF-β downregulation combined with gemcitabine treatment. CONCLUSION: Cellular levels of TGF-β constitute an important factor in gemcitabine resistance and TGF-β silencing might represent a novel and potent strategy for overcoming OSCC resistance to gemcitabine. Show more

Keywords: OSCC, gemcitabine, resistance, TGF-β, lentivirus

DOI: 10.3233/CBM-201456

Citation: Cancer Biomarkers, vol. 29, no. 2, pp. 179-187, 2020

Authors: Ricco, Gabriele | Cosma, Chiara | Bedogni, Giorgio | Biasiolo, Alessandra | Guarino, Maria | Pontisso, Patrizia | Morisco, Filomena | Oliveri, Filippo | Cavallone, Daniela | Bonino, Ferruccio | Plebani, Mario | Brunetto, Maurizia Rossana

Article Type: Research Article

Abstract: BACKGROUND: The time-related variability of HCC biomarkers has not been investigated so far. OBJECTIVE: To assess the changes of alpha-fetoprotein (AFP) and protein induced by vitamin-K absence/antagonist-II (PIVKA-II) in patients with HCC (HCC+ ) as compared to patients without HCC (HCC- ). METHODS: AFP and PIVKA-II were measured by a single laboratory using an automated chemiluminescent-enzyme-immunoassay (Fujirebio Inc., Tokyo, Japan) in 1163 sera of 418 cirrhotics (31.1% HBV, 58.6% HCV, 10.3% non-viral etiology) undergoing ultrasound HCC surveillance. The mean (range) number of effective time-points available for analysis was 2.8 …(2.0 to 3.0); 124 patients with HCC were matched with 294 who remained HCC free for at least 12 months after the last specimen. AFP and PIVKA-II changes were estimated over time by means of a random-effect generalized least squares (RE-GLS) regression model under the missingness at random assumption. RESULTS: Patients with and without HCC had comparable chronic liver disease etiology and staging. AFP/PIVKA-II median (25 th ; 75 th percentile) values at the latest time-point were 4.2 (2.6; 8.6) ng/mL/32 (25; 42) mAU/mL in HCC- and 8.4 (4.4; 32.1) ng/mL/66 (32; 192) mAU/mL in HCC+ (p < 0.001). Log 10 AFP and log 10 PIVKA-II time-changes differed in HCC+ and HCC- patients. In HCC+ patients, both log 10 AFP and log 10 PIVKA-II showed an increasing trend over time. In HCC- patients, log 10 PIVKA-II variations were minimal as compared to log 10 AFP variations. The percent increase of log 10 AFP at 6 months vs . baseline was 11% (95%CI 5 to 17%) and 5% (95%CI 1 to 8%) for log 10 PIVKA-II in HCC+ vs . HCC- patients. CONCLUSIONS: The present retrospective study of the biological variability of AFP and PIVKA-II suggests that their time-related changes may serve as potential predictors of HCC. This topic needs to be addressed by longitudinal studies. Show more

Keywords: AFP, biomarkers, cirrhosis, hepatocellular carcinoma, PIVKA-II, surveillance

DOI: 10.3233/CBM-190118

Citation: Cancer Biomarkers, vol. 29, no. 2, pp. 189-196, 2020

Authors: Yang, Modan | Tan, Winyen | Yang, Xinyu | Zhuo, Jianyong | Lin, Zuyuan | Cen, Beini | Lian, Zhengxing | Li, Huihui | Lu, Di | Wei, Xuyong | Zheng, Shusen | Xu, Xiao

Article Type: Research Article

Abstract: BACKGROUND: Precise recipient selection optimizes the prognosis of liver transplantation (LT) for hepatocellular carcinoma (HCC). Alpha-fetoprotein (AFP) is the most commonly used biomarker for diagnosis and prognosis of HCC in the clinical context. As a crucial molecule in methionine cycle, homocysteine (Hcy) level has been proved to be related to HCC progression and metastasis. OBJECTIVE: We aimed to explore the prognostic capacity of pre-transplant serum Hcy level in LT for HCC. METHODS: This study retrospectively enrolled 161 HCC patients who had underwent LT from donation after cardiac death (DCD) in the First Affiliated …Hospital of Zhejiang University from 2015.01.01 to 2018.09.01. Pre-transplant serum Hcy level was incorporated into statistical analysis together with other clinical parameters and pathological features. RESULTS: From an overall perspective, significant difference was observed in Hcy level between recurrence (n = 61) and non-recurrence group (n = 100) though subsequent analysis showed unsatisfactory predicting performance. In the whole cohort, multivariate analysis showed that lnAFP (p = 0.010) and Milan criteria (MC, p < 0.001) were independent risk factors of HCC recurrence after LT. MA score based on MC and lnAFP performed well in predicting post-LT tumor recurrence with the AUROC at 0.836 (p < 0.001) and 3-year recurrence-free survival rate at 96.8% (p < 0.001) in the low risk group (n = 69). According to the clinical practice, serum concentration lower than 20 μ g/L is considered as normal range of AFP. Elevated pre-transplant serum AFP (> 20 μ g/L) predicts high HCC recurrence after LT. We further divided the 161 recipients into AFP - group (n = 77, AFP ⩽ 20 μ g/L) and AFP + group (n = 84, AFP > 20 μ g/L). MA score was still well presented in the AFP + group and the AUROC for tumor recurrence was 0.823 (p < 0.001), whereas the predicting accuracy was reduced in AFP - group (AUROC: 0.754, P < 0.001). After subsequent analysis, we found that elevated pre-transplant Hcy level (> 12.75 μ mol/L) predicted increased tumor recurrence risk in AFP - group. The 3-year recurrence-free survival rates were 92.0% and 53.7% (p < 0.001) in low Hcy subgroup (n = 40) and high Hcy subgroup (n = 37) respectively. Multivariate analysis showed that Hcy (p = 0.040) and Milan criteria (p = 0.003) were independent risk factors for post-transplant tumor recurrence in AFP - group. Further combination of Hcy level and Milan criteria identified a subgroup of AFP - recipients with acceptable outcomes even though beyond Milan criteria (3-year recurrence-free survival rate: 77.7%, p < 0.001). CONCLUSION: As a classic predictor in HCC prognosis, AFP performed well in our study cohort when combined with Milan criteria. Homocysteine was an effective prognostic biomarker in LT for AFP - hepatocellular carcinoma. In recipients exceeding Milan criteria, acceptable post-transplant outcome could be seen in those with low Hcy and AFP level. Show more

Keywords: Homocysteine, alpha-fetoprotein, Milan criteria, liver transplantation, hepatocellular carcinoma, tumor recurrence

DOI: 10.3233/CBM-201545

Citation: Cancer Biomarkers, vol. 29, no. 2, pp. 197-206, 2020

Authors: Xue, Zhanxia | Gao, Yongshan | Wu, Xueliang

Article Type: Research Article

Abstract: BACKGROUND: Patients with acute leukemia (AL) refractory to induction or reinduction chemotherapy show poor prognoses if they do not undergo allogeneic hematopoietic stem-cell transplantation (AHSCT). The present study aims to investigate whether donor natural killer (NK) cells and interleukin-2 (IL-2) gene modification exert anti-relapse effects on AHSCT after establishing a mouse model of AL. METHODS: C57BL/6 (H-2 b ) mice were selected as donor mice to obtain NK cells and hematopoietic stem cells, while BALB/c (H-2 d ) mice were selected as the recipient mice for AHSCT. The AHSCT-treated mice …were then injected with the donor NK cells, recombinant adenovirus expressing IL-2 (AdIL-2), or the NK cells infected by AdIL-2. Flow cytometry was performed to detect the cell transplantation rate, immune cell number, and cell immunogenicity. Enzyme-linked immunosorbent assay (ELISA) was employed to quantify the secretion of IL-2 in spleen cells, and the level of peripheral blood factors, including interferon-γ (IFN-γ ), tumor necrosis factor-α (TNF-α ), IL-35, transforming growth factor β (TGF-β ), and IL-10. RESULTS: In our experiments, promotional effects of NK cells and AdIL-2 were found on cell transplantation rate, immune reconstitution ability, cell immunogenicity, IL-2 secretion, as well as increased peripheral blood factor levels in the recipient mice treated with AHSCT, with improved pathological changes observed. Moreover, the aforementioned changes were further promoted in the AHSCT-treated recipient mice injected with the AdIL-2-infected NK cells. CONCLUSIONS: These results uncover that the donor NK cells and IL-2 gene modification could inhibit the relapse of AL mice underwent AHSCT, hereby providing a new target for leukemia treatment. Show more

Keywords: Acute leukemia, allogeneic hematopoietic stem cell transplantation, cell immunogenicity, donor natural killer cells, gene modification, interleukin-2

DOI: 10.3233/CBM-191296

Citation: Cancer Biomarkers, vol. 29, no. 2, pp. 207-219, 2020

Authors: Cai, Zeling | Wei, Yi | Chen, Shuai | Gong, Yu | Fu, Yue | Dai, Xianghua | Zhou, Yan | Yang, Haojun | Tang, Liming | Liu, Hanyang

Article Type: Research Article

Abstract: BACKGROUND: Alimentary tract cancers (ATCs) are the most malignant cancers in the world. Numerous studies have revealed the tumorigenesis, diagnosis and treatment of ATCs, but many mechanisms remain to be explored. METHODS: To identify the key genes of ATCs, microarray datasets of oesophageal cancer, gastric cancer and colorectal cancer were obtained from the Gene Expression Omnibus (GEO) database. In total, 207 differentially expressed genes (DEGs) were screened. KEGG and GO function enrichment analyses were conducted, and a protein-protein interaction (PPI) network was generated and gene modules analysis was performed using STRING and Cytoscape. RESULTS: …Five hub genes were screened, and the associated biological processes indicated that these genes were mainly enriched in cellular processes, protein binding and metabolic processes. Clinical survival analysis showed that COL10A1 and KIF14 may be significantly associated with the tumorigenesis or pathology grade of ATCs. In addition, relative human ATC cell lines along with blood samples and tumour tissues of ATC patients were obtained. The data proved that high expression of COL10A1 and KIF14 was associated with tumorigenesis and could be detected in blood. CONCLUSION: In conclusion, the identification of hub genes in the present study helped us to elucidate the molecular mechanisms of tumorigenesis and identify potential diagnostic indicators and targeted treatment for ATCs. Show more

Keywords: Alimentary tract cancers, differentially expressed genes, microarray datasets, enrichment analysis, survival analysis

DOI: 10.3233/CBM-201580

Citation: Cancer Biomarkers, vol. 29, no. 2, pp. 221-233, 2020

Authors: Kim, Sun Il | Koo, Ja Seung

Article Type: Research Article

Abstract: BACKGROUND: Phyllodes tumor (PT) is a rare tumor showing various malignant potential. The histological grade of PT is related to clinical outcome, but its relationship between gaining of malignant potential and underlying mechanism including cancer stem cell factor was not understood yet. OBJECTIVE: The main purpose of this study was to determine the expression pattern of cancer stem cell marker in PT and to understand its clinical and pathological implications. METHODS: CD44, CD166, ALDH1, and Ki-67 immunohistochemistry were performed on a tissue microarray from 185 cases of PT specimens (138 benign, 32 borderline, …15 malignant). The immunohistochemistry result and clinicopathological parameter of each cases were compared to analyze the implications of cancer stem cell markers on PT. RESULTS: Borderline/malignant PT showed higher CD44 expression of the stromal component than benign PT (p < 0.001). In lower histologic grade PT, CD166 showed increased expression in the epithelial component (p = 0.019), but decreased in the stromal component (p < 0.001). Stromal overgrowth was rarely observed as the number of positive cancer stem cell markers increased in the epithelial component (p < 0.001). In the stromal component, the number of positive cancer stem cell markers was related to higher histologic grade (p < 0.001), and increased stromal cellularity (p < 0.001), stromal atypia (p = 0.003), and stromal mitosis (p = 0.002). In benign PT, CD44 negativity (p = 0.013) and a decreased number of positive cancer stem cell markers (p = 0.012) in the epithelial component were related to poor prognosis. CONCLUSIONS: The cancer stem cell markers, CD44 and CD166, are expressed in both the epithelial and stromal components of phyllodes tumor. Besides, ALDH1 is only expressed in stromal component. In the stromal component, expression of cancer stem cell markers increases with higher PT histologic grade. In the epithelial component, the absence of cancer stem cell marker expression is related to poor clinical prognosis. Show more

Keywords: Breast, cancer stem cell, phyllodes tumor

DOI: 10.3233/CBM-191276

Citation: Cancer Biomarkers, vol. 29, no. 2, pp. 235-243, 2020

Authors: Mao, Zhijun | Wang, Zhen | Zhang, Shiping | Pu, Yansong | Wang, Jianhua | Zhang, Tao | Long, Yanbin | Liu, Yi | Ma, Yu | Zhu, Jing

Article Type: Research Article

Abstract: BACKGROUND: Low-density lipoprotein receptor-related protein 4 (LRP4) has been reported to be implicated in multiple types of cancers. However, the significance of LRP4 in gastric cancer (GC) remains poorly elucidated. Therefore, it’s urgent to investigate the importance and underlying mechanisms of LRP4 in GC. OBJECTIVE: To investigate the clinical roles of LRP4 in GC. METHODS: The LRP4 mRNA and miR-140-5p was measured by qRT-PCR. The protein expression was determined Western blot. Kaplan-Meier survival curves and Cox proportional hazard regression models were performed to evaluate prognosis. RESULTS: We demonstrated that LRP4 …mRNA and protein was up-regulated in GC tissues for the first time. Its high expression was significantly correlated with malignant clinical features including TNM stage and lymph-node metastasis and poor prognosis for GC patients. LRP4 promotes migration, invasion and epithelial-mesenchymal transition (EMT) progress of GC cells. Mechanically, LRP4 regulated PI3K/AKT in GC cells. AKT inhibitors reversed the effects of LRP4. Finally, LRP4 was regulated by miR-140-5p in GC. CONCLUSIONS: Our findings showed that LRP4 has an important function in GC progression and promotes GC migration, invasion and EMT by regulating PI3K/AKT under regulation of miR-140-5p, providing a potential therapeutic target for GC. Show more

Keywords: Gastric cancer, LRP4, miR-140-5p, EMT, PI3K/AKT

DOI: 10.3233/CBM-190571

Citation: Cancer Biomarkers, vol. 29, no. 2, pp. 245-253, 2020

Authors: Zhou, Da-Lei | Liu, Qing | Xu, Bo-Heng | Li, Yue | Su, Xuan | Ye, Zu-Lu | Zhang, Xiao | Peng, Jun-Ling | Deng, Ling | Tang, Tao | Shao, Qiong | Ma, Jiang-Jun | Yang, Xin-Hua | He, Cai-Yun

Article Type: Research Article

Abstract: The long non-coding RNA (lncRNA) GAS8-AS1 is the second-most frequently altered gene, following the BRAF gene, in papillary thyroid carcinoma (PTC). We aimed to study the specificity and significance of genetic alterations in GAS8-AS1 in PTC. In this study, we reported the prevalence of genetic alterations of GAS8-AS1 in tissues of 48 nodular goiter, 573 papillary thyroid cancer, 95 colorectal cancer, 101 non-small cell lung cancer, 92 glioma, and 69 gastrointestinal stromal tumor patients, and in peripheral white blood cells of 286 healthy volunteers. We observed that the genomic sequence of GAS8-AS1 had a high …frequency of genetic alterations in addition to the previously reported c.713A> G/714T> C substitution. Substitution of c.713A> G was completely linked with four other loci at c.714T> C, c.728A> G, c.737G> A, and c.752G> A. Two novel substitutions at c.749G> A and c.826A> G were also found. Interestingly, evidence from different samples indicated that these variations were not unique variants for PTC; they were also found in other malignant tissues and white blood cells of healthy volunteers. The c.713A> G substitution was associated with the T stage of PTC, while c.749G> A was more likely to occur in younger patients with PTC. PTC patients carrying heterozygous variants at the c.749 and c.826 loci had a higher risk of developing multiple lesions. These associations were also observed in patients with PTC and concomitant benign thyroid disease. Notably, the rare homozygous GG at the c.826 site conferred a higher risk of developing T2 PTC without benign thyroid disease, and a lower risk of developing T2 PTC with benign thyroid disease. Alterations of c.749G> A and c.826A> G had higher levels of serum TSH (thyroid stimulating hormone) in PTC subjects. Our study provides evidence that the detection of GAS8-AS1 genetic alterations would be useful in diagnostic screening and prognostic assessment of PTC. Show more

Keywords: Papillary thyroid carcinoma, thyroid function, genetic variation, lncRNA

DOI: 10.3233/CBM-191071

Citation: Cancer Biomarkers, vol. 29, no. 2, pp. 255-264, 2020

Authors: Yang, Sen | Wang, Yingshu | Ren, Jun | Zhou, Xueqin | Cai, Kaizhi | Guo, Lijuan | Wu, Shichao

Article Type: Research Article

Abstract: BACKGROUND: Patients with oral squamous carcinoma (OSCC) present difficulty in precise diagnosis and poor prognosis. OBJECTIVE: We aimed to identify the diagnostic and prognostic indicators in OSCC and provide basis for molecular mechanism investigation of OSCC. METHODS: We collected sequencing data and clinical data from TCGA database and screened the differentially expressed mRNAs (DEmRNAs) and lncRNAs (DElncRNAs) in OSCC. Machine learning and modeling were performed to identify the optimal diagnostic markers. In order to determine lncRNAs with prognostic value, survival analysis was performed through combing the expression profiles with the clinical data. Finally, …co-expressed DEmRNAs of lncRNAs were identified by interacted network construction and functional annotated by GO and KEGG analysis. RESULTS: A total of 1114 (345 up- and 769 down-regulated) DEmRNAs and 156 (86 up- and 70 down-regulated) DElncRNAs were obtained in OSCC. Following the machine learning and modeling, 15 lncRNAs were identified to be the optimal diagnostic indicators of OSCC. Among them, FOXD2.AS1 was significantly associated with survival rate of patients with OSCC. In addition, Focal adhesion and ECM-receptor interaction pathways were found to be involved in OSCC. CONCLUSIONS : FOXD2.AS1 might be a prognostic marker for OSCC and our study may provide more information to the further study in OSCC. Show more

Keywords: Oral squamous carcinoma, lncRNA, diagnosis, prognosis, biomarker, machine learning

DOI: 10.3233/CBM-191215

Citation: Cancer Biomarkers, vol. 29, no. 2, pp. 265-275, 2020