Tumor Biology - Volume 44, issue 1

Authors: Piper, Thomas B. | Nielsen, Hans J. | Christensen, Ib Jarle

Article Type: Research Article

Abstract: BACKGROUND: It was previously shown in three subpopulations that subjects not identified with colorectal cancer (CRC) at bowel endoscopy, but with increased serological cancer-associated protein biomarker levels had an increased risk of being diagnosed with subsequent malignant diseases. Objective: The aim of the present study was to perform a pooled analysis of subjects from the three subpopulations and subsequently validate the results in an independent study. The study population denoted the training set includes N = 4,076 subjects with symptoms attributable to CRC and the independent validation set N = 3,774 similar subjects. METHODS: Levels of CEA, CA19-9, TIMP-1 and YKL-40 were …determined in blood samples collected prior to diagnostic bowel endoscopy. Follow-up of subjects not diagnosed with CRC at endoscopy, was ten years and identified subjects diagnosed with primary intra- or extra-colonic malignant diseases. The primary analysis was time to a newly diagnosed malignant disease and was analyzed with death as a competing risk in the training set. Subjects with HNPCC or FAP were excluded. The cumulated incidence was estimated for each biomarker and in a multivariate model. The resulting model was then validated on the second study population. RESULTS: In the training set primary malignancies were identified in 515 (12.6%) of the 4,076 subjects, who had a colorectal endoscopy with non-malignant findings. In detail, 33 subjects were subsequently diagnosed with CRC and 482 subjects with various extra-colonic cancers. Multivariate additive analysis of the dichotomized biomarkers demonstrated that CEA (HR = 1.50, 95% CI:1.21–1.86, p  < 0.001), CA19-9 (HR = 1.41, 95% CI:1.10–1.81, p  = 0.007) and TIMP-1 (HR = 1.25 95% CI: 1.01–1.54, p  = 0.041) were significant predictors of subsequent malignancy. The cumulated incidence at 5 years landmark time was 17% for those subjects with elevated CEA, CA19-9 and TIMP-1 versus 6.7% for those with low levels of all. When the model was applied to the validation set the cumulated 5-year incidence was 10.5% for subjects with elevated CEA, CA19-9 and TIMP-1 and 5.6% for subjects with low levels of all biomarkers. Further analysis demonstrated a significant interaction between TIMP-1 and age in the training set. The age dependency of TIMP-1 indicated a greater risk of malignancy in younger subjects if the biomarker was elevated. This observation was validated in the second set. CONCLUSION: Elevated cancer-associated protein biomarker levels in subjects with non-malignant findings at large bowel endoscopy identifies subjects at increased risk of being diagnosed with subsequent primary malignancy. CEA, CA19-9 and TIMP-1 were significant predictors of malignant disease in this analysis. TIMP-1 was found dependent on age. The results were validated in an independent symptomatic population. Show more

Keywords: Colorectal cancer, colorectal neoplasia, malignancy, protein biomarkers, endoscopy

DOI: 10.3233/TUB-211501

Citation: Tumor Biology, vol. 44, no. 1, pp. 1-16, 2022

Authors: Atoum, Manar F. | Alzoughool, Foad E. | Al-Mazaydeh, Zainab A. | Rammaha, Majdoleen S. | Tahtamouni, Lubna H.

Article Type: Research Article

Abstract: BACKGROUND: 1,25-dihydroxyvitamin D3 (1,25(OH)2 D3 ) is an effective anticancer agent, and when combined with other agents it shows superior activities. Vitamin B12 has been shown to contribute to increasing the effectiveness of anticancer drugs when used in combination. Thus, the current study aimed at investigating the anticancer potential of the combination of 1,25(OH)2 D3 and vitamin B12. METHODS: MTT assay was used to determine the cytotoxic activity of combining 1,25(OH)2 D3 and vitamin B12 against six different cancer cell lines and one normal cell line. The surviving fraction after clonogenic assay was measured, and …the effects of 1,25(OH)2 D3 /B12 combination on the activity of different caspases, cell adhesion, actin cytoskeleton, cell morphology, and percentage of polarized cells were evaluated. RESULTS: Vitamin B12 did not cause cytotoxicity, however, it enhanced the cytotoxicity of 1,25(OH)2 D3 against cancer cells. The cytotoxic effects of 1,25(OH)2 D3 and its combination with vitamin B12 was not evident in the normal mammary MCF10A cell line indicating cancer cell-specificity. The cytotoxic effects of 1,25(OH)2 D3 /B12 combination occurred in a dose-dependent manner and was attributed to apoptosis induction which was mediated by caspase 4 and 8. Moreover, 1,25(OH)2 D3 /B12-treated cells showed enhanced inhibition of clonogenic tumor growth, reduced cell adhesion, reduced cell area, reduced percentage of cell polarization, and disorganized actin cytoskeleton resulting in reduced migratory phenotype when compared to cells treated with 1,25(OH)2 D3 alone. CONCLUSION: 1,25(OH)2 D3 and vitamin B12 exhibited synergistic anticancer effects against different cancer cell lines. The combination therapy of 1,25(OH)2 D3 and vitamin B12 may provide a potential adjunctive treatment option for some cancer types. Show more

Keywords: 1α,25-Dihydroxyvitamin D3, cobalamin, combined therapy, synergistic effects, ER stress

DOI: 10.3233/TUB-211536

Citation: Tumor Biology, vol. 44, no. 1, pp. 17-35, 2022

Authors: Kasurinen, Jussi | Hagström, Jaana | Kaprio, Tuomas | Beilmann-Lehtonen, Ines | Haglund, Caj | Böckelman, Camilla

Article Type: Research Article

Abstract: BACKGROUND: A large number of infiltrating CD3- and CD8-positive inflammatory cells indicates an improved survival in colorectal cancer (CRC), similar to many other cancers. OBJECTIVE: We investigated the prognostic value of different combinations of CD3- and CD8-positive immune cells in CRC patients. METHODS: The densities of CD3- and CD8-positive cells in intratumoral and stromal tissues were evaluated from 539 patients, for which we calculated a CD3 tumor–stroma index, a CD8 tumor–stroma index, and a CD3–CD8 tumor–stroma index. RESULTS: High CD3 and CD8 tumor–stroma indices associated with stage I to II disease (p  <  0.001 …for both). The CD3 tumor–stroma index associated with a colonic tumor location (p  = 0.006), while the CD8 tumor–stroma index associated with right-sided tumors (p  <  0.001) and histological grade 3 tumors (p  = 0.032). High intratumoral and stromal densities for CD3- and CD8-positive immune cells, the CD3 tumor–stroma index, the CD8 tumor–stroma index, and the CD3–CD8 tumor–stroma index all indicated a better DSS. CONCLUSIONS: The CD3 tumor–stroma index carries a strong prognostic value in CRC, and none of the CD3 and CD8 combinations we analyzed proved superior. Show more

Keywords: CD3, CD8, T-lymphocytes, colorectal cancer, immunohistochemistry

DOI: 10.3233/TUB-211571

Citation: Tumor Biology, vol. 44, no. 1, pp. 37-52, 2022

Authors: Algharib, Ali Samy | Shanab, Gamila Mohamed-Labib | Abdel-Ghaffar, Abdel-Rahman Badr | Ismail, Mohamed Ahmed | Mohamed, Rania Hassan

Article Type: Research Article

Abstract: BACKGROUND: Bithiophene derivatives show a promising anti-cancer potential. We previously showed that Bithienyl Fluorobenzamidine (BFB) has an anti-proliferative effect against several leukemia cell lines. Acute myeloid leukemia (AML) accounts for 18% of the total leukemia cases worldwide with heavier burden during the past 30 years. Therefore, the main aim remains the discovery of safe and effective medications. OBJECTIVE: The current research aims to investigate the anti-cancer efficacy of BFB and its effect on the apoptosis in the 7,12-Dimethylbenz[a]anthracene (DMBA) induced AML in mice. METHODS: AML was induced in mice by DMBA and then …treated by 2 different doses of BFB. After BFB treatment, the hematological and histological pattern changes was examined. Furthermore, the molecular effect of BFB on apoptosis, cell cycle markers and Protein kinase B (Akt) pathway was examined using qPCR, Western blotting and ELISA. RESULTS: BFB treatment ameliorates leukemia histological and hematological markers significantly, despite non-significant changes in normal mice. This improvement exhibits cell cycle arrest and apoptosis induction, represented by elevation of tp53 /p53, p21 /p21, Caspase3 and downregulation of ckk1 /Cdk1 in the bone marrow, as well as Akt pathway suppression. CONCLUSIONS: Our results establishes BFB as a promising therapeutic candidate against AML through cell cycle arrest, apoptosis induction and Akt pathway modulation. Show more

Keywords: Acute myeloid leukemia, bithienyl fluorobenzamidine, cell cycle, apoptosis, Akt pathway

DOI: 10.3233/TUB-211538

Citation: Tumor Biology, vol. 44, no. 1, pp. 53-67, 2022

Authors: Eurola, Annika | Ristimäki, Ari | Mustonen, Harri | Nurmi, Anna-Maria | Hagström, Jaana | Kallio, Pauliina | Alitalo, Kari | Haglund, Caj | Seppänen, Hanna

Article Type: Research Article

Abstract: BACKGROUND: Wnt/β-catenin signaling is a highly conserved signaling pathway that regulates the transcription factor PROX1. The role of β-catenin and PROX1 in pancreatic cancer is ambiguous, as some studies have associated their expression with tumor regression and some with tumor progression. OBJECTIVE: We have investigated their expression in surgically treated pancreatic cancer patients receiving neoadjuvant therapy (NAT), and patients treated upfront with surgery (US). We furthermore compared the expression of β-catenin and PROX1 between patients who had a good or poor response to NAT. METHODS: We evaluated β-catenin and PROX1 expression through immunohistochemistry in 88 neoadjuvant …and 144 upfront surgery patients by scoring the intensity of the immunopositivity as 0–3, corresponding to negative, weak, moderate, or strong. We developed a six-tier grading scheme for the neoadjuvant responses by analyzing the remaining tumor cells in surgical specimen histological sections. RESULTS: Strong β-catenin immunopositivity associated with improved survival in the patients with good NAT-response (≤10% residual tumor cells) (Hazard ratio [HR] 0.26 95%, confidence interval [CI] 0.07–0.88 p  = 0.030). Additionally, the combined moderate β-catenin and PROX1 expression associated with improved survival (HR 0.20 95% CI 0.05–0–76 p  = 0.018) among the good responders. Among the patients with a poor NAT-response (> 10% residual tumor cells), both strong β-catenin immunopositivity and strong combined β-catenin and PROX1 associated with shorter survival (HR 2.03 95% CI 1.16–3.55 p  = 0.013, and HR 3.1 95% CI 1.08–8.94 p  = 0.03, respectively). PROX1 alone was not associated with survival. CONCLUSIONS: Strong β-catenin immunopositivity and combined strong or moderate β-catenin and PROX1 immunopositivity associated with improved survival among the good NAT-responders and worse survival among the poor NAT-responders. Show more

Keywords: Pancreatic ductal adenocarcinoma, PDAC, preoperative chemotherapy, treatment response, chemoresistance, tumor microenvironment

DOI: 10.3233/TUB-211581

Citation: Tumor Biology, vol. 44, no. 1, pp. 69-84, 2022

Authors: Demarchi, Gianina | Valla, Sofía | Perrone, Sofía | Chimento, Agustina | Bonadeo, Nadia | Vitale, Daiana Luján | Spinelli, Fiorella Mercedes | Cervio, Andrés | Sevlever, Gustavo | Alaniz, Laura | Berner, Silvia | Cristina, Carolina

Article Type: Research Article

Abstract: INTRODUCTION: Prolactinomas are the most frequent pituitary tumor subtype. Despite most of them respond to medical treatment, a proportion are resistant and become a challenge in clinical management. Wnt/β-Catenin pathway has been implicated in several cancers including pituitary tumors and other sellar region malignancies. Interestingly, Wnt/β-Catenin inhibition augments the cytotoxicity of the chemotherapeutic agent Temozolomide (TMZ) in different cancers. TMZ is now being implemented as rescue therapy for aggressive pituitary adenoma treatment. However, the molecular mechanisms associated with TMZ action in pituitary tumors remain unclear. OBJECTIVES: Our aims in the present study were to evaluate differential β-Catenin expression …in human resistant prolactinomas and Wnt/β-Catenin signaling activation and involvement in Prolactin (PRL) secreting experimental models treated with TMZ. RESULTS: We first evaluated by immunohistochemistry β-Catenin localization in human resistant prolactinomas in which we demonstrated reduced membrane β-Catenin in prolactinoma cells compared to normal pituitaries, independently of the Ki-67 proliferation indexes. In turn, in vivo 15 mg/kg of orally administered TMZ markedly reduced PRL production and increased prolactinoma cell apoptosis in mice bearing xenografted prolactinomas. Intratumoral β-Catenin strongly correlated with Prl and Cyclin D1 , and importantly, TMZ downregulated both β-Catenin and Cyclin D1, supporting their significance in prolactinoma growth and as candidates of therapeutic targets. When tested in vitro , TMZ directly reduced MMQ cell viability, increased apoptosis and produced G2 /M cell cycle arrest. Remarkably, β-Catenin activation and VEGF secretion were inhibited by TMZ in vitro . CONCLUSIONS: We concluded that dopamine resistant prolactinomas undergo a β-Catenin relocalization in relation to normal pituitaries and that TMZ restrains experimental prolactinoma tumorigenicity by reducing PRL production and β-Catenin activation. Together, our findings contribute to the understanding of Wnt/β-Catenin implication in prolactinoma maintenance and TMZ therapy, opening the opportunity of new treatment strategies for aggressive and resistant pituitary tumors. Show more

Keywords: Prolactinomas, temozolomide, β-Catenin, pituitary tumors, Wnt

DOI: 10.3233/TUB-211500

Citation: Tumor Biology, vol. 44, no. 1, pp. 85-105, 2022

Authors: Niture, Suryakant | Tricoli, Lucas | Qi, Qi | Gadi, Sashi | Hayes, Kala | Kumar, Deepak

Article Type: Research Article

Abstract: OBJECTIVES: MicroRNAs (miRNAs) are the small non-coding regulatory RNA molecules involved in gene regulation via base-pairing with complementary sequences in mRNAs. The dysregulation of specific miRNAs, such as miR-99b-5p (miR-99b), is associated with prostate cancer (PCa) progression. However, the mechanistic role of miR-99b in PCa remains to be determined. In this study, we aimed to investigate the functional and clinical significance of miR-99b in PCa. STUDY DESIGN: The expression of miR-99b and its downstream targets mTOR/AR in the PCa samples were analyzed by RT/qPCR. The effects of miR-99b overexpression/inhibition on PCa cell survival/proliferation, spheroid formation, and cell migration …were examined by specific assays. Luciferase reporter assays were performed to determine the binding of miR-99b to 3′ untranslated region (UTR) of the mTOR gene. The effects of miR-99b on the expression of mTOR, AR, and PSA proteins, as well as on AKT/mTOR signaling, autophagy, and neuroendocrine differentiation markers were analyzed by western blotting. The expression of miR-99b, mTOR, AR, PSA in AR-negative PC3 and AR-positive LNCaP cells was analyzed by RT/qPCR. The effect of miR-99b on global gene expression in PC3 cells was analyzed by RNA-seq. RESULTS: The expression of miR-99b was downregulated in tumor samples from PCa patients, whereas the expression of mTOR and AR was upregulated. In PCa cell lines, overexpression of miR-99b inhibited cell proliferation and cell colony/spheroid formation; induced apoptosis, and increased sensitivity towards docetaxel (DTX). In contrast, inhibition of miR-99b by miR-99b inhibitor resulted in increased cell growth in PCa cells. Mechanistically, miR-99b inhibited the expression of the mammalian target of the rapamycin (mTOR) gene by binding to its 3′ UTR and induced autophagy. Furthermore, miR-99b inhibited androgen receptor (AR) activity in LNCaP cells and induced apoptosis. Activation of AR signaling by dihydrotestosterone (DHT) downregulated miR-99b expression and promoted cell PCa cell growth/survival, whereas inactivation of mTOR by rapamycin or AR by enzalutamide decreased miR-99b mediated PCa cell growth. CONCLUSION: Our data suggest that miR-99b functions as a tumor suppressor by targeting the mTOR/AR axis in PCa cells, implicating miR-99b as a novel biomarker and therapeutic target for PCa management. Show more

Keywords: miR-99b-5p, mTOR, AR, cell survival, autophagy, apoptosis, prostate cancer

DOI: 10.3233/TUB-211568

Citation: Tumor Biology, vol. 44, no. 1, pp. 107-127, 2022

Authors: Manukonda, Radhika | Attem, Jyothi | Yenuganti, Vengala Rao | Kaliki, Swathi | Vemuganti, Geeta K.

Article Type: Research Article

Abstract: Exosomes are a subgroup of membrane-bound extracellular vesicles secreted by all cell types and present virtually in all biological fluids. The composition of exosomes in the same cell type varies in healthy and disease conditions. Hence, exosomes research is a prime focus area for clinical research in cancer and numerous age-related metabolic syndromes. Functions of exosomes include crucial cell-to-cell communication that mediates complex cellular processes, such as antigen presentation, stem cell differentiation, and angiogenesis. However, very few studies reported the presence and role of exosomes in normal physiological and pathological conditions of specialized ocular tissues of the eye and ocular …cancers. The eye being a protected sense organ with unique connectivity with the rest of the body through the blood and natural passages, we believe that the role of exosomes in ocular tissues will significantly improve our understanding of ocular diseases and their interactions with the rest of the body. We present a review that highlights the existence and function of exosomes in various ocular tissues, their role in the progression of some of the neoplastic and non-neoplastic conditions of the eyes. Show more

Keywords: Exosomes, ocular tumors, extracellular vesicles, clinical applications

DOI: 10.3233/TUB-211543

Citation: Tumor Biology, vol. 44, no. 1, pp. 129-152, 2022

Authors: Jafarpour, Sima | Yazdi, Maryam | Nedaeinia, Reza | Ghobakhloo, Sepideh | Salehi, Rasoul

Article Type: Research Article

Abstract: INTRODUCTION: Controversy exists regarding the association of apolipoprotein B mRNA editing enzyme catalytic subunit 3B APOBEC3B , (A3B ) overexpression and poor prognosis, metastasis, and chemotherapy drug resistance in cancers. Here we conducted a systematic review and meta-analysis to determine its prognostic value and clinicopathological features in breast cancer and some other malignancies. MATERIALS AND METHODS: PubMed, Scopus, Cochrane Library, Web of Science, and EMBASE were searched up to Feb 2022 for the association of A3B with breast, ovarian, gastrointestinal and lung cancers. The pooled hazard ratios with 95% confidence interval (CI) were evaluated to assess disease-free …survival (DFS), overall survival (OS), and recurrence-free survival (RFS) in cancers under study. RESULTS: Over 3700 patients were included in this meta-survey. Elevated levels of A3B were significantly related to low OS (pooled HR = 1.30; 95% CI:1.09–1.55, P  < 0.01), poor DFS (pooled HR = 1.66; 95% CI:1.17–2.35, P  < 0.01) and poor RFS (HR = 1.51, 95% CI:1.11–2.04, P  = 0.01). Subgroup analysis revealed that high A3B expression was associated with poor OS in lung (HR = 1.85, 95% CI: 1.40–2.45), and breast cancers (HR = 1.38, 95% CI: 1.00–1.89). High expression of A3B did not display any significant association with clinicopathologic features. CONCLUSION: APOBEC3B overexpression is related to poor OS, DFS and RFS only in some cancer types and no generalized role could be predicted for all cancers. Show more

Keywords: APOBEC3B, A3B, overall survival, prognosis, biomarker

DOI: 10.3233/TUB-211577

Citation: Tumor Biology, vol. 44, no. 1, pp. 153-169, 2022

Authors: Manganaro, Lucia | Celli, Veronica | Viggiani, Valentina | Berardelli, Elena | Granato, Teresa | Tartaglione, Sara | Farina, Antonella | Catalano, Carlo | Angeloni, Antonio | Anastasi, Emanuela

Article Type: Research Article

Abstract: BACKGROUND: Hereditary ovarian cancers (HOC) represent about 23% of ovarian cancer (OC) cases: they are most frequently related to germline mutations in the BRCA genes. OBJECTIVE: We aimed to compare CA125/HE4 serum levels and Computed Tomography (CT) features at time of ovarian cancer (OC) diagnosis in two populations: BRCA mutant and BRCA wild-type (WT) OC, and to investigate the relationship between this laboratory and radiological biomarker and BRCA mutation status. METHODS: This retrospective study included 60 newly diagnosed OC patients with FIGO stage IIIC-IV disease, tested for BRCA1/2 germline mutation status of which preoperative CT scan …and serum tumor marker assay were available. RESULTS: The median level of CA125 (708 U/mL) was significantly higher (p  < 0.002) in BRCA1/2 mutated patients than in WT patients (176 U/mL), whereas the median level of HE4 (492 pmol/L) was significantly higher (p  < 0.002) in WT than in BRCA-mutated patients (252 pmol/L). BRCA mutation carriers showed a higher incidence of bilateral ovarian masses (p  = 0.0303) characterized by solid structures (p  < 0.00001), higher peritoneal tumor load, macronodular implants >2 cm (p  = 0.000099), increased frequency of lymphadenopathies (p  = 0.019), and metastasis (p  = 0.052) compared to patients with BRCA WT. CONCLUSIONS: Tumor markers and CT patterns may help in identifying BRCA mutation status in OC directing patients towards a personalized treatment. Show more

Keywords: Ovarian cancer, BRCA, computed tomography, HE4-CA125

DOI: 10.3233/TUB-211557

Citation: Tumor Biology, vol. 44, no. 1, pp. 171-185, 2022