Cancer Biomarkers - Volume 32, issue 3

Authors: Li, Min | Zhu, Yilong | Bai, Bing | Fang, Jinbo | Yao, Wei | Li, Yiquan | Li, Shanzhi | Li, Xiao | Jin, Ningyi | Jiang, Rihua

Article Type: Research Article

Abstract: BACKGROUND: To explore the suppressive effect of Apoptin-loaded oncolytic adenovirus (Ad-VT) on luciferase-labeled human melanoma cells in vitro and in vivo . METHODS: The stable luciferase-expressing human melanoma cells A375-luc or M14-luc were obtained by transfecting the plasmid pGL4.51 and selection with G418, followed by luciferase activity, genetic stability and bioluminescence intensity assays. In vitro , the inhibitory effects of Ad-VT on A375-luc or M14-luc were evaluated using the MTS cell proliferation, FITC-Annexin V apoptosis detection, transwell migration, Matrigel invasion and scratch assays. The inhibition pathway in Ad-VT-infected A375-luc and M14-luc cells were analyzed by …JC-1 staining and Western-blot detection of mitochondrial apoptosis-related proteins. In vivo , the suppressive effects of Ad-VT on A375-luc or M14-luc were assessed by living imaging technology, tumor volume, bioluminescence intensity, survival curves and immunohistochemical analysis of the tumors from the xenograft tumor model BALB/c nude mice. RESULTS: The growth and migration of A375-luc and M14-luc were significantly inhibited by Ad-VT in vitro . The evaluations of A375-luc and M14-luc tumor models in BALB/c nude mice were successfully performed using living imaging technology. Ad-VT had an anti-tumor effect by reducing tumor growth and increasing survival in vivo . Ad-VT significantly changed the mitochondrial membrane potential by triggering the the mitochondrial release of apoptosis-related proteins, AIF (apoptosis inducing factor), ARTS (Apoptosis-Related Proteins), and Cyto-c (cytochrome c) from the mitochondria. CONCLUSION: Ad-VT reduced the mitochondrial membrane potential in A375-luc or M14-luc cells and induced the mitochondrial release of AIF, ARTS and Cyto-C. Ad-VT induced apoptosis in A375-luc or M14-luc cells via the mitochondrial apoptotic pathway. Show more

Keywords: Luciferase labeled A375 or M14, living imaging technology, oncolytic adenovirus, tumor suppression, BALB/C nude mice model

DOI: 10.3233/CBM-203150

Citation: Cancer Biomarkers, vol. 32, no. 3, pp. 251-262, 2021

Authors: Aref, Salah | Ebrahim, Lamiaa | El-Ashwah, Shaimaa | El Agdar, Mohamed | Ayed, Mohamed

Article Type: Research Article

Abstract: BACKGROUND: Acute myeloid leukemia changes the bone marrow (BM) niche to support leukemia cells by modulating the stromal microenvironment. The aim is to assess Activin-A as a biomarker in acute myeloid leukemia (AML). METHODS: The level of Activin-A and CXCL-12 protein concentration levels in the plasma of bone marrow aspirate samples of eighty AML patients at diagnosis, after induction and at relapse were determined by ELISA. RESULTS: We found that Activin-A concentration levels was significantly up regulated in AML cases at diagnosis, and down regulated at complete remission and rise again at relapse …(P < 0.001). In contrast; the CXCL-12 gene expression was significantly down regulated in AML cases at diagnosis; relapse, and up regulated after complete remission (P < 0.001). Multivariate analysis showed that high Activin-A levels at diagnosis is significant predictor of induction of remission response OR 1.006 (CI: 1.002–1.010) (P = 0.003); AML relapse OR 1.002 (CI: 1.0–1.004) (P = 0.043) as well as patients’ outcome OR 1.33 (CI: 1.004–1.062) (P = 0.024). CONCLUSION: Activin-A level at diagnosis is a new simple easily assessed biomarker that could predict AML patient’s response to therapy as well as patient’s outcome. Show more

Keywords: Activin-A, CXCL-12, AML

DOI: 10.3233/CBM-203171

Citation: Cancer Biomarkers, vol. 32, no. 3, pp. 263-270, 2021

Authors: Li, Yuxin | Zhuang, Xiaohong | Zhuang, Li | Liu, Hongjian

Article Type: Research Article

Abstract: This paper aimed at investigating AS1 expression in prostate cancer (PCa) and its effects on the proliferation and invasion of prostate cancer cells (PCCs). The prostate tissues and the matched adjacent normal prostate tissues excised and preserved during radical prostatectomy in our hospital were collected. The LncRNA NCK1-AS1 expression was detected. PCa patients were followed up for three years to analyze their prognosis. The correlation of LncRNA NCK1-AS1 expression with clinicopathological features was analyzed. Human normal prostate cells and human PCCs were selected, in which LncRNA NCK1-AS1 expression was tested to screen and then transfect the cells. Cell proliferation, invasion …and migration were detected. Cell cycles and apoptosis were analyzed. Compared with the adjacent normal tissues, LncRNA NCK1-AS1 was highly expressed in the prostate cancer tissues. Its expression was remarkably different in those with different stages of TNM and with lymphatic metastasis or not. The prognosis of patients with high LncRNA NCK1-AS1 expression was remarkably poorer than that of those with low expression. Compared with the human normal prostate cells, LncRNA NCK1-AS1 expression in the human PCCs remarkably rose, with the greatest difference in 22Rv1 cells. Compared with the Blank group, cell proliferation and the number of plate cloned cells remarkably reduced in the sh-NCK1-AS1 group. Additionally, in this group, the number of invasive and migratory cells remarkably reduced; the expression of invasion-related protein E-cadherin remarkably rose but that of MMP-2 remarkably reduced; cell cycles were arrested and the expression of cycle-related proteins (CDK4, CDK6, cyclin D1) remarkably reduced; the apoptotic rate and the expression of apoptosis-related protein Bax remarkably rose. LncRNA NCK1-AS1 is highly expressed in PCa, so its down-regulation can inhibit PCCs from proliferating and reduce the number of invasive cells. Show more

Keywords: LncRNA NCK1-AS1, prostate cancer, proliferation, invasion, migration, cell cycle

DOI: 10.3233/CBM-203021

Citation: Cancer Biomarkers, vol. 32, no. 3, pp. 271-279, 2021

Authors: Homkham, Nontiya | Muangwong, Pooriwat | Pisprasert, Veeradej | Traisathit, Patrinee | Jiratrachu, Rungarun | Chottaweesak, Pattawee | Chitapanarux, Imjai

Article Type: Research Article

Abstract: BACKGROUND: Immune-enhancing nutrition (IMN) strengthens the systematic inflammatory response and the immune system. Neutrophil to lymphocyte ratio (NLR) and absolute lymphocyte count (ALC) are affected during cancer therapies. OBJECTIVE: We carried out an analysis of the dynamic changes in NLR and ALC over time in cancer patients with or without IMN supplementation. METHODS: 88 cancer patients receiving concurrent chemoradiotherapy (CCRT) were randomized into regular diet group, and regular diet and IMN group.Generalized estimation equation models were used to assess associations between patient’s characteristics, IMN, and dynamic changes in NLR and ALC over time. …RESULTS: NLR and ALC at pre-CCRT were significantly associated with dynamic changes in NLR (adjusted β = 1.08, 95% confidence interval [CI]: 0.64–1.52) and ALC (adjusted β = 0.41, 95% CI: 0.36–0.46). The magnitudes of the NLR and ALC changes through CCRT were lower in patients receiving IMN, although the differences were not statistically significant except ALC at the end of CCRT in head and neck cancer patients (P = 0.023). CONCLUSION: Dynamic negative changes in both markers were demonstrated throughout CCRT. There were non-significant trend in promising changes in both NLR and ALC values in the whole group in IMN supplementation. Show more

Keywords: Concurrent chemoradiotherapy, immune-enhancing nutrition, neutrophil to lymphocyte ratio, absolute lymphocyte count, cancer

DOI: 10.3233/CBM-210086

Citation: Cancer Biomarkers, vol. 32, no. 3, pp. 281-291, 2021

Authors: Guan, Lirong | Zhang, Lingli | Wang, Tengqi | Jia, Lizhou | Zhang, Ning | Yan, Huishan | Zhao, Kun

Article Type: Research Article

Abstract: BACKGROUND: Nuclear pore membrane protein 121 (POM121) is a novel biomarker involved in tumorigenesis and metastasis. However, little is known about the role of POM121 in non-small-cell lung cancer (NSCLC). OBJECTIVE: The aim of this study was to detect the expression of POM121 in NSCLC and its relationship with clinicopathologic feature and cell biological behavior, and explore the underlying mechanisms. METHODS: The expression of POM121 in NSCLC tissues and para-carcinoma tissues was compared by quantitative real-time PCR and immunohistochemistry analysis. The relationship between POM121 protein and clinicopathological characteristics in NSCLC was investigated. Roles …of POM121 in NSCLC cells were investigated by CCK-8 assay, clone formation assay, transwell migration and invasion assay, and in vivo experiments. Variations of signaling pathways were determined by qRT-PCR and Western blot. RESULTS: The POM121 expression in NSCLC tissues was significantly higher than that in para-carcinoma tissues, both at the mRNA and protein level. The POM121 expression was related to sex, advanced differentiation, tumor diameter, lymph node metastases, distant metastases, American Joint Committee on Cancer (AJCC) stage, venous invasion, and perineural invasion in NSCLC. Kaplan-Meier analysis indicated that NSCLC patients with high POM121 expression had poor overall survival. Downregulation of POM121 inhibited cell proliferation, clone formation, migration and invasion. TGF-β /SMAD and PI3K/AKT pathways were involved in POM121-induced functional changes in NSCLC cells. CONCLUSION: POM121 plays an oncogenic role in NSCLC through TGF-β /SMAD and PI3K/AKT pathways. POM121 expression is a potential independent prognostic factor for NSCLC. Show more

Keywords: POM121, non-small-cell lung cancer, prognosis, TGF-β/SMAD, PI3K/AKT

DOI: 10.3233/CBM-210001

Citation: Cancer Biomarkers, vol. 32, no. 3, pp. 293-302, 2021

Authors: Tu, Simei | Zhang, Hao | Yang, Xiaocheng | Wen, Wen | Song, Kangjing | Yu, Xinyi | Qu, Xinjian

Article Type: Research Article

Abstract: BACKGROUND: Since the molecular mechanisms of cervical cancer (CC) have not been completely discovered, it is of great significance to identify the hub genes and pathways of this disease to reveal the molecular mechanisms of cervical cancer. OBJECTIVE: The study aimed to identify the biological functions and prognostic value of hub genes in cervical cancer. METHODS: The gene expression data of CC patients were downloaded from the Gene Expression Omnibus (GEO) database and The Cancer Genome Atlas (TCGA) database. The core genes were screened out by differential gene expression analysis and weighted gene …co-expression network analysis (WGCNA). R software, the STRING online tool and Cytoscape software were used to screen out the hub genes. The GEPIA public database was used to further verify the expression levels of the hub genes in normal tissues and tumour tissues and determine the disease-free survival (DFS) rates of the hub genes. The protein expression of the survival-related hub genes was identified with the Human Protein Atlas (HPA) database. RESULTS: A total of 64 core genes were screened, and 10 genes, including RFC5 , POLE3 , RAD51 , RMI1 , PALB2 , HDAC1 , MCM4 , ESR1 , FOS and E2F1 , were identified as hub genes. Compared with that in normal tissues, RFC5 , POLE3 , RAD51 ,RMI1 , PALB2 , MCM4 and E2F1 were all significantly upregulated in cervical cancer, ESR1 was significantly downregulated in cervical cancer, and RFC5 expression in CC patients was significantly related to OS. In the DFS analysis, no significant difference was observed in the expression level of RFC5 in cervical cancer patients. Finally, RFC5 protein levels verified by the HPA database were consistently upregulated with mRNA levels in CC samples. CONCLUSIONS: RFC5 may play important roles in the occurrence and prognosis of CC. It could be further explored and validated as a potential predictor and therapeutic target for CC. Show more

Keywords: Cervical cancer, gene differential expression analysis, weighted gene co-expression network analysis, hub gene, biomarkers

DOI: 10.3233/CBM-203262

Citation: Cancer Biomarkers, vol. 32, no. 3, pp. 303-315, 2021

Authors: Kabzinski, Jacek | Maczynska, Monika | Kaczmarczyk, Dariusz | Majsterek, Ireneusz

Article Type: Research Article

Abstract: BACKGROUND: Reduced efficiency of DNA repair systems has long been a suspected factor in increasing the risk of cancer. OBJECTIVE: In this work we investigate influence of three selected polymorphisms of DNA repair gene XRCC1 and level of oxidative damage (measured as level of 8-oxo-guanine) on modulation of the risk of HNSCC. METHODS: In group of 359 patients with HNSCC (diagnosed with OSCC) the occurrence of polymorphic variants in Arg399Gln, Arg280His and Arg194Trp of XRCC1 were studied with TaqMan technique. In addition we determined level of 8-oxo-guanine with ELISA. …RESULTS: Arg399Gln polymorphism and Arg194Trp polymorphism of XRCC1 gene increases the risk of HNSCC. The coexistence of Arg399Gln and Arg194Trp simultaneously enhances this effect. At the same time, their coexistence with His280His raises the risk to a level higher than in the absence of such coexistence, although the His280His itself is not associated with an increased risk of HNSCC. Patients have higher levels of 8-oxo-guanine than control group, and His280His is polymorphism with highest mean value of 8-oxoG level among studied. CONCLUSION: Patients with HNSCC not only have an increased level of 8-oxoguanine and the Arg399Gln and Arg/Trp of XRCC1 modulate risk of cancer, but there is also a relationship between these two phenomena, and it can be explained using intragenic combinations revealing that a high level of 8-oxoG could be a potential mechanism behind the modulation of HNSCC risk by the polymorphisms studied. Show more

Keywords: HNSCC, OSCC, DNA repair, oxidative stress, XRCC1

DOI: 10.3233/CBM-203163

Citation: Cancer Biomarkers, vol. 32, no. 3, pp. 317-326, 2021

Authors: Metwally, Ayman Mohamed | Li, Hanchen | Houghton, JeanMarie

Article Type: Research Article

Abstract: BACKGROUND: Recent studies demonstrated the involvement of mesenchymal stem/stromal cells (MSCs) in carcinogenesis, but the molecular mechanism behind this transformation is still obscured. OBJECTIVE: To screen both the expression levels of polycomb and trithorax epigenetic regulators and TrP53 mutations in early and late MSC culture passages in an attempt to decipher the mechanism of spontaneous transformation. METHODS: The study was conducted on early and late passages of MSC culture model from C57BL/6J mice. The expression profile of 84 epigenetic regulators was examined using RT2 profiler PCR array. TrP53 mutations in the DNA binding …domain was screened. Codons, amino acids positions and the corresponding human variants were detected in P53 sequences. RESULTS: Sixty-two epigenetic regulators were dysregulated. Abnormalities were detected starting the third passage. Nine regulators were dysregulated in all passages. (C> G) substitution P53 mutation was detected in passage 3 resulting in Ser152Arg substitution. Passages 6, 9, 12 and the last passage showed T> C substitution resulting in Cys235Arg substitution. The last passage had T deletion and A insertion resulting in frame shift mutations changing the p.Phe286Ser and p.Asn103Lys respectively. CONCLUSION: In vitro expanded MSCs undergo transformation through alteration of epigenetic regulators which results in genomic instability and frequent P53 mutations. Show more

Keywords: MSCs, epigenetic regulators, P53, mutation, transformation, C57BL/6J mice

DOI: 10.3233/CBM-203121

Citation: Cancer Biomarkers, vol. 32, no. 3, pp. 327-337, 2021

Authors: Gan, Feng-Jiao | Li, Yi | Xu, Meng-Xi | Zhou, Tie | Wu, Shun | Hu, Kang | Li, Yan | Sun, Su-Hong | Luo, Qing

Article Type: Research Article

Abstract: BACKGROUND: Neoadjuvant chemotherapy (NAC) is an important treatment for locally advanced breast cancer (LABC). However, there are no effective biomarkers to predict the efficacy. Therefore, there is an urgent need for new biomarkers to predict the response of LABC to NAC. LncRNA BCAR4 has been detected in a variety of malignant tumor tissues and used as a new biomarker for diagnosis and prognosis. However, LncRNA BCAR4 predicts the response of LABC to NAC is unclear. OBJECTIVE: Explore the predictive effect of LncRNA BCAR4 on the efficacy of NAC for LABC in three different evaluation systems. …METHODS: First, the TCGA database was used to analyze the expression of LncRNA BCAR4 in 33 kinds of malignant tumors, and further explore its expression in breast cancer and its impact on the survival and prognosis of breast cancer. Furthermore, quantitative methods were used to measure the expression level of LncRNA BCAR4 in cancer tissues of 48 LABC patients, and the correlation between LncRNA BCAR4 and clinicopathological status and response to NAC under the evaluation system of 3, RECIST1.1, Miller-Payne (MP) score and whether it reaches pCR,was analyzed. RESULTS: TCGA data analysis found that LncRNA is highly expressed in a variety of malignant tumor tissues, including breast cancer. And relatively low expression, the shorter the overall survival time of high expression patients. The high expression of LncRNA BCAR4 is related to the size of the tumor, and there are differences in expression between stage I and other stages, but there is no obvious correlation with the positive lymph node and hormone receptor status. Among the three evaluation systems, only in the RECIST 1.1 evaluation system LncRNA BCAR4 has a predictive effect on NAC for LABC. The expression of LncRNA BCAR4 has no significant correlation with clinical stage, Ki-67% and hormone receptor status, and has no significant correlation with whether patients with locally advanced breast cancer obtain pCR during neoadjuvant chemotherapy. CONCLUSION: LncRNA BCAR4 is highly expressed in LABC tissues and may be an effective marker for predicting the efficacy of NAC for LABC. Show more

Keywords: Locally advanced breast cancer, neoadjuvant chemotherapy, LncRNA BCAR4, pathological complete response, clinical complete response

DOI: 10.3233/CBM-210048

Citation: Cancer Biomarkers, vol. 32, no. 3, pp. 339-351, 2021

Authors: Temesfői, Viktória | Molnár, Kinga | Kaltenecker, Péter | Réger, Barbara | Szomor, Árpád | Horváth-Szalai, Zoltán | Alizadeh, Hussain | Kajtár, Béla | Kőszegi, Tamás | Miseta, Attila | Nagy, Tamás | Faust, Zsuzsanna

Article Type: Research Article

Abstract: BACKGROUND: Recent studies proved that metabolic changes in malignant disorders have an impact on protein glycosylation, however, only a few attempts have been made so far to use O-GlcNAc analysis as a prognostic tool. Glucose metabolism is reported to be altered in hematological malignancies thus, we hypothesized that monitoring intracellular O-GlcNAc levels in Rai stage 0-I (Binet A) CLL patients could give deeper insights regarding subtle metabolic changes of progression which are not completely detected by the routine follow-up procedures. OBJECTIVE: In this proof of concept study we established a flow cytometric detection method for the …assessment of O-GlcNAcylation as a possible prognostic marker in CLL malignancy which was supported by fluorescence microscopy. METHODS: Healthy volunteers and CLL patients were recruited for this study. Lymphocytes were isolated, fixed and permeabilised by various methods to find the optimal experimental condition for O-GlcNAc detection by flow cytometry. O-GlcNAc levels were measured and compared to lymphocyte count and various blood parameters including plasma glucose level. RESULTS: The protocol we developed includes red blood cell lysis, formalin fixation, 0.1% Tween 20 permeabilisation and employs standardized cell number per sample and unstained controls. We have found significant correlation between O-GlcNAc levels and WBC (R 2 = 0.8535, p < 0.0029) and lymphocyte count (R 2 = 0.9225, p < 0.0006) in CLL patients. Interestingly, there was no such correlation in healthy individuals (R 2 = 0.05664 for O-GlcNAc vs WBC and R 2 = 0.04379 for O-GlcNAc vs lymphocytes). CONCLUSION: Analyzing O-GlcNAc changes in malignant disorders, specifically in malignant hematologic diseases such as CLL, could be a useful tool to monitor the progression of the disease. Show more

Keywords: Chronic lymphocytic leukemia, O-GlcNAcylation, RL2, immunometabolism, flow cytometry

DOI: 10.3233/CBM-203049

Citation: Cancer Biomarkers, vol. 32, no. 3, pp. 353-362, 2021