Multistep Carcinogenesis Research Articles

Anti-VEGFR2 neutralising antibody slows the progression of multistep oral carcinogenesis.

Angiogenesis plays an important role in cancer growth and metastasis, and it is considered a therapeutic target to control tumour growth following anti-angiogenic therapy. However, it is still unclear when tissues initiate angiogenesis during malignant transformation from premalignant condition and whether this premalignant condition could be a therapeutic target of anti-angiogenic therapy. In this study, we aimed to analyse the onset of angiogenesis by evaluating morphological and functional alterations of microvessels during oral multistep carcinogenesis using a 4-nitroquinoline 1-oxide (4NQO)-induced oral carcinogenesis mouse model. In the study, we initially confirmed that with the use of 4NQO, oral lesions develop in a stepwise manner from normal mucosa through oral epithelial dysplasia (OED) to oral squamous cell carcinoma (OSCC). Evaluation of CD31-immunostained specimens revealed that microvessel density (MVD) increases in a stepwise manner from OEDs. Histological and functional analyses revealed the structural abnormalities and leakage of blood vessels had already taken place in OED. Then we evaluated the expression profiles of Hif1a and Vegfa along with hypoxic status and found that OED exhibited increased Vegfa expression under hypoxic conditions. Finally, we tested the possibility of OEDs as a target of anti-angiogenic therapy and found that anti-VEGFR2 neutralising antibody in OED slowed the disease progression from OED to OSCC. These data indicate that an angiogenic switch occurs at the premalignant stage and morphological, and functional alterations of microvessels already exist in OED. These findings also elucidate the tumour microenvironment, which gradually develops along with carcinogenic processes, and highlight usefulness of the 4NQO-induced carcinogenesis model in the study of epithelial and stromal components, which will support epithelial carcinogenesis. © 2024 The Author(s). The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Artemisinin pre-treatment fore cisplatin dosage enhances high grade urothelial carcinoma treatment in male albino mice via reverse gene expression modulation of FGFR3, HRAS, P53 and KDM6A

BackgroundUrinary bladder cancer, is the 10th most common global cancer, diagnosed in over 600,000 people causing 200,000 deaths annually. Artemisinin and its derivatives are safe compounds that have recently been proven to possess potent anti-tumor effects in vivo, through inhibition of cancer cell growth. The aim of this study is to assess the efficiency of artemisinin as a cancer treatment alone and as a pre-treatment fore cisplatin therapy for high grade urothelial carcinoma.MethodsSixty male albino mice were divided into six groups, and BBN was used to induce urinary bladder cancer. Blood samples were tested for renal functions and complete blood counts, kidney and urinary bladder tissues were harvested for histopathological examination. Total RNAs from urinary bladder tissues was collected, and gene expression of FGFR3, HRAS, P53, and KDM6A was quantified using qRT-PCR.ResultsCompared to the induced cancer group, the results revealed that FGFR3 expression levels were down-regulated in the induced cancer group treated by artemisinin only and the induced cancer group pre-treated with artemisinin prior to cisplatin by ~ 0.86-fold and 0.4-folds, respectively, aligning with HRAS down-regulation by ~ 9.54-fold and 9.05-fold, respectively. Whereas, P53 expression levels were up-regulated by ~ 0.68-fold and 0.84-fold, respectively, in parallel with KDM6A expression, which is up-regulated by ~ 0.95-folds and 5.27-folds, respectively. Also, serum creatinine and urea levels decreased significantly in the induced cancer group treated by artemisinin alone and the induced cancer group pre-treated with artemisinin prior to cisplatin, whereas the induced cancer group treated by cisplatin their levels increased significantly. Moreover, Hb, PLT, RBC, and WBC counts improved in both cancer groups treated by artemisinin alone and pre-treated with artemisinin prior to cisplatin. Histologically, in kidney tissues, artemisinin pre-treatment significantly reduced renal injury caused by cisplatin. While Artemisinin treatment for cancer in bladder tissues reverted invasive urothelial carcinoma to moderate urothelial dysplasia.ConclusionsThis study indicates that artemisinin demonstrated a significant effect in reversal of the multi-step carcinogenesis process of high grade urothelial carcinoma and could enhance the effect of cisplatin therapy using artemisinin pre-treatment.

Establishment and Characterization of Amitrole-Induced Mouse Thyroid Adenomatous Nodule-Derived Cell Lines.

Background: Thyroid cancer cell lines have been of great value for the study of thyroid cancer. However, the availability of benign thyroid adenoma cell lines is limited. Methods: Cell lines were established from thyroid adenomatous nodules that developed in mice treated with the goitrogen amitrole. Expression of epithelial, mesenchymal, and thyroid markers of these established cell lines was determined, and the effect of lentivirus-transduced overexpression of NKX2-1, a master regulator of thyroid development, on the thyroid marker expression was examined. Signal transduction and cell proliferation were evaluated after treatment with insulin-like growth factor-I (IGF-I) and the selective IGF-I receptor (IGF-IR) inhibitor NVP-ADW742. Xenograft studies were performed to examine tumorigenicity of the cells in mice. Whole-genome sequencing (WGS) was used to comprehensively determine the genetic mutations in the established two cell lines. Results: Five mouse thyroid adenomatous nodules-derived cell lines named CAT (cells from amitrole-treated thyroids) were established. Among these, two cell lines, CAT458/458s (CAT458s: a subline of CAT458) and CAT459, were found to be positive for epithelial markers and negative for a mesenchymal marker. NKX2-1-positive CAT459 cells showed higher messenger RNA (mRNA) expression of some thyroid differentiation markers than NKX2-1-negative CAT458s cells, and NKX2-1 overexpression increased and/or induced their expression. IGF-I signaling was transduced in thyrotropin receptor (Tshr)-negative CAT458s and 459 cells, and NVP-ADW742 suppressed their proliferation. No tumors developed in mice after subcutaneous injection of CAT458s or 459 cells. The WGS analysis revealed the presence of missense mutations in the tumor suppressor genes such as Polk (encoding DNA polymerase kappa) and Tgfb1 (encoding transforming growth factor beta 1), while no mutations were found in the prominent thyroid cancer-related genes Braf, Trp53 (encoding p53), and Tert (encoding telomerase reverse transcriptase). Conclusions: Two mouse thyroid adenomatous nodule-derived cell lines with different thyroid differentiation marker expression were established. NKX2-1 induced partial differentiation of these cell lines. They lacked tumorigenicity and prominent gene mutations involved in thyroid cancer development, while missense mutations were found in some tumor suppressors as revealed by WGS. The CAT458s and 459 provide a new tool to further clarify the process of thyroid multistep carcinogenesis and differentiation.

An overview of risk assessment and monitoring of malignant transformation in cirrhotic nodules

Cirrhotic liver nodules can progress to hepatocellular carcinoma (HCC) through a multi-step carcinogenesis model, with dysplastic nodules being particularly high risk. Currently, monitoring the progression of non-HCC cirrhotic nodules is primarily through dynamic observation, but there is a lack of sensitive, efficient, and convenient methods. Dynamic monitoring and risk evaluation of malignant transformation are essential for timely treatment and improved patient survival rates. Routine liver biopsies are impractical for monitoring, and imaging techniques like ultrasound, computed tomography, and magnetic resonance imaging are not suitable for all patients or for accurately assessing subcentimeter nodules. Identifying serum biomarkers with high sensitivity, specificity, and stability, and developing a multi-index evaluation model, may provide a more convenient and efficient approach to monitoring pathological changes in cirrhotic nodules.

The relationships between Risk Factors and the changes of Histopathological of Tissue Colon Cancer in Iraq Patients

The transformation of an ordinary cell into a cancerous one could be a dynamic process which often involves the approval of oncogenes as well as deactivation of tumor suppressor properties (alterations). Most changes are emptied by cellular genome dauntlessness and repair components. In any case, many genetic changes give cells an advancement advantage and unused capacities, driving to their neoplastic alter. As other cancers, like melanoma, sarcoma, and lymphoma, could affect the colon, colon cancer, also known as colon carcinoma, affects the epithelium of colon. The uncontrolled proliferation regarding colorectal epithelial cells might be a hallmark of colorectal cancer (CRC). The multistep carcinogenesis hypothesis states that pigment cells go through a sequence of atomic alterations before they become totally dangerous cells. The inactivation related to the Adenomatous polyposis coli (APC) signaling pathway is the most frequent hereditary cause of activation. The pathology of damage includes oncogene changes, tumor suppressor qualities, and probably many signaling pathways, which put cancers at risk of metastasis. In advanced nations, CRC has a high disease-specific mortality rate and impacts over a million individuals annually. Worldwide, CRC ranks 3 rd in terms of the incidence in both women and men. Africa and Central and South Asia have experienced the biggest drops in CRC, whereas New Zealand, Australia, North America, and Europe have experienced highest change rates.

Liver Lesions at Risk of Transformation into Hepatocellular Carcinoma in Cirrhotic Patients: Hepatobiliary Phase Hypointense Nodules without Arterial Phase Hyperenhancement.

Recent technical advances in liver imaging and surveillance for patients at high risk for developing hepatocellular carcinoma (HCC) have led to an increase in the detection of borderline hepatic nodules in the gray area of multistep carcinogenesis, particularly in those that are hypointense at the hepatobiliary phase (HBP) and do not show arterial phase hyperenhancement. Given their potential to transform and advance into hypervascular HCC, these nodules have progressively attracted the interest of the scientific community. To date, however, no shared guidelines have been established for the decision management of these borderline hepatic nodules. It is therefore extremely important to identify features that indicate the malignant potential of these nodules and the likelihood of vascularization. In fact, a more complete knowledge of their history and evolution would allow outlining shared guidelines for their clinical-surgical management, to implement early treatment programs and decide between a preventive curative treatment or a watchful follow-up. This review aims to summarize the current knowledge on hepatic borderline nodules, particularly focusing on those imaging features which are hypothetically correlated with their malignant evolution, and to discuss current guidelines and ongoing management in clinical practice.

MiR-150 and miR-155 expression predicts survival of cervical cancer patients: a translational approach to novel prognostic biomarkers

IntroductionHigh-risk human papillomavirus (HPV) is the aetiological agent of cervical cancer, which remains the fourth leading cause of cancer death in women worldwide. K14-HPV16 transgenic mice are a model for HPV-induced cancers, which undergo multistep squamous carcinogenesis at the skin, that is histologically and molecularly similar to carcinogenesis of the human cervix. Previous screens of differentially regulated microRNAs (miRs) using K14-HPV16 mice showed a role for miR-21, miR-155, miR-150, miR-146a, miR-125b and miR-223 during carcinogenesis.MethodsWe now aim to translate these observations into the clinical setting, using data provided by The Cancer Genome Atlas (TCGA) to explore whether those microRNAs can influence the survival of cervical cancer patients.ResultsResults showed that low miR-150, miR-155 and miR-146a expression levels in primary tumours were associated with poor overall survival. However, only miR-150 and miR-155 were found to be independent predictors, increasing the risk of death. When patients were stratified by clinical stage, low miR-150, miR-155, miR-146a and miR-125b were associated with poor survival for clinical stages I and II. Only low miR-150 expression increased the death risk.ConclusionWe conclude that miR-150 and miR-155 may be potentially applied as prognostic biomarkers in cervical cancer patients. However, further investigation is required to determine their applicability.

Molecular genetic bases and pathways of carcinogenesis in the pancreas

Pancreatic cancer is a highly aggressive disease with a high mortality rate in need of innovative diagnostic and therapeutic approaches. In recent years, due to the development of molecular genetic methods a large amount of data has appeared on the genetic and epigenetic alterations that occur in pancreatic cancer, and the genetic landscape of this disease has been determined. Additionally, the genetic factors underlying the multistep carcinogenesis in precursor lesions of the pancreas have been clarified. This review highlights the pathways of carcinogenesis in the pancreas, as well as the molecular genetic processes underlying it.

Stepwise activation of p63 and the MEK/ERK pathway induces the expression of ARL4C to promote oral squamous cell carcinoma cell proliferation.

Carcinogenesis is a multistep process wherein cells accumulate multiple genetic alterations and progress to a more malignant phenotype. It has been proposed that sequential accumulation of gene abnormalities in specific genes drives the transition from non-tumorous epithelia through a preneoplastic lesion/benign tumor to cancer. Histologically, oral squamous cell carcinoma (OSCC) progresses in multiple ordered steps that begin with mucosal epithelial cell hyperplasia, which is followed by dysplasia, carcinoma in situ and invasive carcinoma. It is therefore hypothesized that genetic alteration-mediated multistep carcinogenesis would be involved in the development of OSCC; however, the detailed molecular mechanisms are unknown. We clarified the comprehensive gene expression patterns and carried out an enrichment analysis using DNA microarray data from a pathological specimen of OSCC (including a non-tumor region, carcinoma in situ lesion and invasive carcinoma lesion). The expression of numerous genes and signal activation were altered in the development of OSCC. Among these, the p63 expression was increased and the MEK/ERK-MAPK pathway was activated in carcinoma in situ lesion and in invasive carcinoma lesion. Immunohistochemical analyses revealed that p63 was initially upregulated in carcinoma in situ and ERK was sequentially activated in invasive carcinoma lesions in OSCC specimens. ADP-ribosylation factor (ARF)-like 4c (ARL4C), the expression of which is reportedly induced by p63 and/or the MEK/ERK-MAPK pathway in OSCC cells, has been shown to promote tumorigenesis. Immunohistochemically, in OSCC specimens, ARL4C was more frequently detected in tumor lesions, especially in invasive carcinoma lesions, than in carcinoma in situ lesions. Additionally, ARL4C and phosphorylated ERK were frequently merged in invasive carcinoma lesions. Loss-of-function experiments using inhibitors and siRNAs revealed that p63 and MEK/ERK-MAPK cooperatively induce the expression of ARL4C and cell growth in OSCC cells. These results suggest that the stepwise activation of p63 and MEK/ERK-MAPK contributes to OSCC tumor cell growth through regulation of ARL4C expression.

Abstract 1219: Establishment and characterization of amitrole-induced mouse thyroid adenoma-derived cell lines

Abstract Some types of thyroid cancers are known to arise from thyroid adenomas, as a malignant transition from benign adenoma, as proposed in the multi-step carcinogenesis model. Thyroid cancer studies using human thyroid cancer cell lines have been intensely performed by many researchers. However, mouse thyroid adenoma cell lines have not been well established and characterized. In order to obtain further insights into thyroid carcinogenesis process, we established five cell lines from thyroid adenomas developed in mice treated with goitrogen amitrole. Among them, two cell lines, CAT458/458s (CAT: cells from amitrole treated thyroids; CAT458s: a sub-line from CAT458) and CAT459 were found to be positive for E-cadherin and Claudin-1, representative epithelial markers, and negative for Vimentin, a mesenchymal marker, suggesting that they originated from thyroid epithelial cells. We found higher expression of DUOX1, a member of NADPH oxidase involved in thyroid hormone synthesis, in both CAT458s and 459 cells than normal mouse thyroids, presumably due to an unknown mechanism caused by amitrole treatment. NKX2-1, a master regulator of thyroid development, is not expressed in CAT458s cells while NKX2-1 is abundantly expressed in CAT459 cells. Consistent with this finding, NKX2-1-positive CAT459 cells showed higher mRNA expression of thyroid differentiation markers including Tg, Duox1, and Duox2. NKX2-1 overexpression in NKX2-1-negative CAT458s cells using a lentiviral infection system induced mRNA expressions of these thyroid differentiation marker genes, suggesting that partial thyroid differentiation of the adenoma cells is induced by NKX2-1. TSH and IGF-I are known to work synergistically for thyroid growth. However, Tshr mRNA expression is not detected in either CAT458s or 459 cells, suggesting that TSH signal is not responsible for their growth. On the other hand, IGF-IR protein is expressed and IGF-I induces AKT and S6 phosphorylation in both cells. We further tried to assess somatic gene mutations involved in adenoma development by whole-genome sequencing (WGS). WGS analysis revealed that somatic missense mutations are not found in genes known to be involved in thyroid carcinogenesis and cancer progression, such as Braf, Trp53, Ret, Tert, Hras, Kras, and Nras. This finding suggests that prominent oncogenic mutations are not required for thyroid adenoma development that may later progress to carcinoma. Currently we are validating several missense mutations, including mutations in tumor suppressor genes Atm and Tgfb1, found in these cell lines. In conclusion, we established mouse thyroid adenoma cell lines with different differentiation and molecular profiles. CAT458s and 459 cells would provide a tool to further clarify the process of thyroid adenoma development and thyroid differentiation. Citation Format: Yo-Taro Shirai, Nobuo Hoshi, Huaitian Liu, Maxwell P. Lee, Shioko Kimura. Establishment and characterization of amitrole-induced mouse thyroid adenoma-derived cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1219.

Targeting epigenetic features in clear cell sarcomas based on patient-derived cell lines

BackgroundClear cell sarcomas (CCSs) are translocated aggressive malignancies, most commonly affecting young adults with a high incidence of metastases and a poor prognosis. Research into the disease is more feasible when adequate models are available. By establishing CCS cell lines from a primary and metastatic lesion and isolating healthy fibroblasts from the same patient, the in vivo process is accurately reflected and aspects of clinical multistep carcinogenesis recapitulated.MethodsIsolated tumor cells and normal healthy skin fibroblasts from the same patient were compared in terms of growth behavior and morphological characteristics using light and electron microscopy. Tumorigenicity potential was determined by soft agar colony formation assay and in vivo xenograft applications. While genetic differences between the two lineages were examined by copy number alternation profiles, nuclear magnetic resonance spectroscopy determined arginine methylation as epigenetic features. Potential anti-tumor effects of a protein arginine n-methyltransferase type I (PRMT1) inhibitor were elicited in 2D and 3D cell culture experiments using cell viability and apoptosis assays. Statistical significance was calculated by one-way ANOVA and unpaired t-test.ResultsThe two established CCS cell lines named MUG Lucifer prim and MUG Lucifer met showed differences in morphology, genetic and epigenetic data, reflecting the respective original tissue. The detailed cell line characterization especially in regards to the epigenetic domain allows investigation of new innovative therapies. Based on the epigenetic data, a PRMT1 inhibitor was used to demonstrate the targeted antitumor effect; normal tissue cells isolated and immortalized from the same patient were not affected with the IC50 used.ConclusionsMUG Lucifer prim, MUG Lucifer met and isolated and immortalized fibroblasts from the same patient represent an ideal in vitro model to explore the biology of CCS. Based on this cell culture model, novel therapies could be tested in the form of PRMT1 inhibitors, which drive tumor cells into apoptosis, but show no effect on fibroblasts, further supporting their potential as promising treatment options in the combat against CCS. The data substantiate the importance of tailored therapies in the advanced metastatic stage of CCS.

COX-2 Expression as a Prognostic Inflammatory Marker in Head and Neck Squamous Cell Carcinoma: A Cross-sectional Study

Introduction: Head and Neck Squamous Cell Carcinoma (HNSCC) ranks as the sixth leading cause of cancer worldwide. HNSCC is an aggressive neoplasm involving multistep carcinogenesis. As it is associated with an increased risk of recurrence, morbidity and mortality, many studies are being undertaken to understand its pathophysiology at the molecular level, identify newer prognostic factors and develop novel therapeutic agents. Aim: To assess the clinicopathological features of all cases of HNSCC, including age, sex, site and risk factors and to demonstrate the expression of the Cyclooxygenase-2 (COX-2) marker in various grades of the tumour in a subset of cases. Materials and Methods: A cross-sectional study was conducted on HNSCC at the Institute of Pathology, Rajiv Gandhi Government General Hospital, Chennai, Tamil Nadu, India for a period of one year from July 2014 to June 2015. A total of 50 cases (representing different grades) were randomly selected, 25 each from the oral cavity/oropharynx and upper respiratory tract. Immunohistochemistry (IHC) was performed using the COX-2 marker and the results were tabulated in an MS Excel sheet to assess the incidence among different age groups, genders, various risk factors, and sites. Statistical analysis was performed using Statistical Package for the Social Sciences (SPSS) 21.0 software, using the Chi-square test to assess the significant expression of COX-2 in varying grades of the tumour. A p-value <0.05 was considered significant. Results: The study results (total cases=50) showed an increased incidence of HNSCC among males (n=39; 78%) compared to females (n=11; 22%). Additionally, HNSCC most commonly affected patients in the 5th to 6th decade (n=24; 48%) of life. The occurrence was most commonly noted in the tongue (n=8; 16%) and hypopharynx (n=13; 26%) among all Squamous Cell Carcinoma (SCC) cases of the oral cavity and upper respiratory tract, respectively. Out of the 50 cases included to analyse COX-2 expression, 41 cases (82%) of HNSCC showed COX-2 expression, and its association with different grades was evident by a significant p-value of 0.027. Conclusion: This study demonstrated significant expression of COX-2 in all grades of HNSCC, supporting the role of inflammation in multistep carcinogenesis. Furthermore, molecular targeted therapy against COX-2 could be implemented as an adjunctive therapy with chemoradiation to hinder the progression and aggressive behaviour of the tumour.

Stem Cell Theory of Cancer: Origin of Metastasis and Sub-clonality.

Metastasis may be the secret weapon cancer uses to dominate and subjugate, to persist and prevail. However, it is no longer a secret when we realize that a stem cell has the same ways and means to fulfill its own omnipotence and accomplish its own omnipresence… and when we realize that a cancer cell has its own version of stem-ness origin and stem-like nature. In this perspective, we discuss whether stem-ness enables metastasis or mutations drive metastasis. We ponder about low-grade versus high-grade tumors and about primary versus metastatic tumors. We wonder about stochasticity and hierarchy in the genesis and evolution of cancer and of metastasis. We postulate that metastasis may hold the elusive code that makes or breaks a stem-cell versus a genetic theory of cancer. We speculate that the vaunted model of multistep carcinogenesis may be in error and needs some belated remodeling and a major overhaul. We propose that subsequent malignant neoplasms from germ cell tumors and donor-derived malignancies in organ transplants are quintessential experiments of nature and by man that may eventually empower us to elucidate a stem-cell origin of cancer and metastasis. Unfortunately, even the best experiments of cancer and of metastasis will be left unfinished, overlooked, or forgotten, when we do not formulate a proper cancer theory derived from pertinent and illuminating clinical observations. Ultimately, there should be no consternations when we realize that metastasis has a stem-cell rather than a genetic origin, and no reservations when we recognize that metastasis has been providing us some of the most enduring tests and endearing proofs to demonstrate that cancer is indeed a stem-cell rather than a genetic disease after all.

Detecting Posttranslational Modifications of Hsp90 Isoforms.

The molecular chaperone heat shock protein 90 (Hsp90) is essential in eukaryotes. Hsp90 chaperones proteins that are important determinants of multistep carcinogenesis. There are multiple Hsp90 isoforms including the cytosolic Hsp90α and Hsp90β as well as GRP94 located in the endoplasmic reticulum and TRAP1 in the mitochondria. The chaperone function of Hsp90 is linked to its ability to bind and hydrolyze ATP. Co-chaperones and posttranslational modifications (such as phosphorylation, SUMOylation, and ubiquitination) are important for Hsp90 stability and regulation of its ATPase activity. Both mammalian and yeast cells can be used to express and purify Hsp90 and TRAP1 and also detect post-translational modifications by immunoblotting.

Genomic profiling and network-level understanding uncover the potential genes and the pathways in hepatocellular carcinoma.

Data integration with phenotypes such as gene expression, pathways or function, and protein-protein interactions data has proven to be a highly promising technique for improving human complex diseases, particularly cancer patient outcome prediction. Hepatocellular carcinoma is one of the most prevalent cancers, and the most common cause is chronic HBV and HCV infection, which is linked to the majority of cases, and HBV and HCV play a role in multistep carcinogenesis progression. We examined the list of known hepatocellular carcinoma biomarkers with the publicly available expression profile dataset of hepatocellular carcinoma infected with HCV from day 1 to day 10 in this study. The study covers an overexpression pattern for the selected biomarkers in clinical hepatocellular carcinoma patients, a combined investigation of these biomarkers with the gathered temporal dataset, temporal expression profiling changes, and temporal pathway enrichment following HCV infection. Following a temporal analysis, it was discovered that the early stages of HCV infection tend to be more harmful in terms of expression shifting patterns, and that there is no significant change after that, followed by a set of genes that are consistently altered. PI3K, cAMP, TGF, TNF, Rap1, NF-kB, Apoptosis, Longevity regulating pathway, signaling pathways regulating pluripotency of stem cells, Cytokine-cytokine receptor interaction, p53 signaling, Wnt signaling, Toll-like receptor signaling, and Hippo signaling pathways are just a few of the most commonly enriched pathways. The majority of these pathways are well-known for their roles in the immune system, infection and inflammation, and human illnesses like cancer. We also find that ADCY8, MYC, PTK2, CTNNB1, TP53, RB1, PRKCA, TCF7L2, PAK1, ITPR2, CYP3A4, UGT1A6, GCK, and FGFR2/3 appear to be among the prominent genes based on the networks of genes and pathways based on the copy number alterations, mutations, and structural variants study.

Understanding the role of molecular and genetic alterations in proliferation and progression of oral squamous cell carcinoma : A review article

Oral carcinogenesis is known as multifactorial process which engaged plentiful genetic events that transform normal activity of tumor suppressor genes and oncogenes. It is observed that aggregation of genetic alterations is the ground for advancement of a normal cell to cancer cells, which is known as a multi-step carcinogenesis. Because of this event, growth factor production increases as well as increase in total of receptors on cell surface and increased intracellular signal messengers. The present review scrutinize the existing documentation in the literature related to the oral squamous cell carcinoma. English language articles were searched in various databases such as Pubmed, Scopus, Science direct and Google scholar. The keyword used for searching are “oral squamous cell carcinoma”, “Genetics and Oral squamous cell carcinoma”,“Molecular mechanism in oral squamous cell carcinoma”. The present review spotlights on understanding the molecular mechanism and the genetic factors which is responsible for alteration in the cell which leads to oral squamous cell carcinoma.

P1301: BIOLOGICAL SIGNIFICANCE OF UCK2 IN HTLV-1-INFECTED CELLS IN ATL LEUKEMOGENESIS AND ACQUIRED RESISTANCE TO AZACITIDINE

Background: Adult T-cell leukemia-lymphoma (ATL) is an aggressive hematological malignancy of CD4+ T-cells transformed by human T-cell lymphotropic virus-1 (HTLV-1). We have reported that regional DNA hypermethylation in HTLV-1-infected T-cells reflects the disease status of ATL and the anti-ATL effects of DNA demethylating agents, azacitidine (AZA) and decitabine (Blood 2020, 136: 871-884). We recently generated AZA-resistant (AZA-R) cells from ATL cell lines via long-term drug exposure and found down-regulation of uridine cytidine kinase2 (UCK2) correlated with lower susceptibility to AZA (Int J Cancer 2022, 150: 1184-1197). UCK1 and UCK2 are involved in mono-phosphorylation of uridine and cytidine in pyrimidine nucleotide biosynthesis. Both proteins also catalyze mono-phosphorylation of AZA. Overexpression of UCK2 has been observed in several types of solid tumor tissues. Aims: Here, we aimed to understand the role of UCK2 in HTLV-1-infected cells during multi-step carcinogenesis of ATL and the biological significance of inactivation of UCK2 in AZA-R cells. Methods: All studies using human samples were performed in accordance with the guidelines set out in the Declaration of Helsinki and were approved by the institutional review board at Saga University (2018-03-02). Peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood samples from healthy volunteers, HTLV-1 carriers and patients with ATL. ATL cell lines were cultured in RPMI-1640 medium containing 10% FBS. Results: We found that UCK2 gene expression was higher in HTLV-1-infected T-cells than in normal T-cells using the gene expression datasets GSE55851 and GSE33615. UCK2 protein expression was increased in HTLV-1-infected T-cells (CADM1+ cells) isolated from patients with ATL compared with uninfected T-cells (Fig. A). We also found UCK2 gene expression was upregulated via activation of T-cell receptor (TCR) signaling pathway in normal CD4+ T-cells (Fig. B). Somatic alterations and epigenetic modifications activating TCR signaling have been identified in ATL cells. Therefore, we think UCK2 protein is overexpressed through dysregulated TCR signaling in HTLV-1-infected cells during ATL leukemogenesis. We have successfully established AZA-R cell lines, whose UCK2 expression was lost or decreased. Although they proliferated normally in vitro, the knockdown of UCK2 dramatically suppressed cell growth of ATL cell lines (Fig. C). Since UCK2 is involved in pyrimidine nucleotide biosynthesis, we then performed metabolome analysis in AZA-R cells. Although uridine and cytidine were accumulated in AZA-R cells, possibly due to loss of UCK2, the amount of UTP and CTP was almost same with parental cells. On the other hands, dihydroorotic acid (DHO) was decreased and orotate was increased in AZA-R cells (Fig. D). DHODH catalyzes the oxidation of DHO to orotic acid and is a rate-limiting enzyme in de novo pyrimidine nucleotide synthesis. AZA-R cells were more susceptible to BAY2402234, a DHODH inhibitor (Fig. E), indicating the activation of pyrimidine de novo synthesis in AZA-R cells. Image:Summary/Conclusion: Taken together, we think UCK2 supports the vigorous cell proliferation not only in normal TCR-stimulated T-cells but also in HTLV-1-infected cells. Inactivation of UCK2 makes cells resistant to AZA but suppresses the cell growth possibly due to the pyrimidine nucleotide starvation. AZA-R cells with reduced UCK2 expression proliferate normally in vitro by the activation of pyrimidine de novo synthesis.

The Immunohistochemical Expression of SOX-10 in Urothelial Carcinoma and the Non Neoplastic Urothelium; and a Correlation with the Tumor Features.

Objective:Evaluation of SOX-10 expression in malignant urothelial cells, comparing it with the phenotypically non neoplastic urothelium, and correlating it with the various clinicopathological variables, with a focus on the invasive pattern. Methods:Eighty paraffin blocks of urothelial carcinoma were stained by H&E. Histopathological features were evaluated and then immunostained with SOX-10 to evaluate its expression. Results:The evaluation of SOX-10 expression in urothelial carcinoma, revealed a high grade of SOX-10 expression in the malignant urothelium (43\\80 cases; 53.7%), while the adjacent the non neoplastic urothelium expressed high SOX-10 in (12\\42 case; 28.6%). Correlation of SOX-10 score with the various variables revealed a statistically significant correlation with the gross shape (P value=0.002), the tumor grade ((P value=0.009), the muscle invasion by the tumor ((P value=0.004), the tumor T stage, (P-value <0.001), N stage (P value=0.003), associated Schistoma hematobium infestation (P-value =0.016), and the presence of vascular tumor emboli (P-value =0.009). It was statistically insignificant with the gender, the anatomical site, and the perineural tumor invasion. Correlating the mean of SOX-10 score with some tumor features revealed a statistically significant correlation with the muscle invasion by the tumor, Tumor grade, T stage, and non neoplastic urothelium; P-value <0.001 each and N stage P value=0.006. Conclusion:SOX-10 is overexpressed in urothelial carcinoma and it was also detected in a significant part of the surrounding non neoplastic urothelium, which may contribute to understanding its role in multistep urothelial carcinogenesis as transcription or tumor-promoting factor, thus it could be used in future trials for specific targeted therapy.

Pathological, molecular, and clinical characteristics of cholangiocarcinoma: A comprehensive review.

Cholangiocarcinomas are a heterogeneous group of highly aggressive cancers that may arise anywhere within the biliary tree. There is a wide geographical variation with regards to its incidence, and risk-factor associations which may include liver fluke infection, primary sclerosing cholangitis, and hepatolithiasis amongst others. These tumours are classified into intrahepatic, perihilar and distal based on their anatomical location. Morphologically, intrahepatic cholangiocarcinomas are further sub-classified into small and large duct variants. Perihilar and distal cholangiocarcinomas are usually mucin-producing tubular adenocarcinomas. Cholangiocarcinomas develop through a multistep carcinogenesis and are preceded by dysplastic and in situ lesions. While clinical characteristics and management of these tumours have been extensively elucidated in literature, their ultra-structure and tumour biology remain relatively unknown. This review focuses on the current knowledge of pathological characteristics, molecular alterations of cholangiocarcinoma, and its precursor lesions (including biliary intraepithelial neoplasia, intraductal papillary neoplasms of the bile duct, intraductal tubulopapillary neoplasms and mucinous cystic neoplasm).

Single-cell RNA Sequencing Uncovered the Involvement of an Endothelial Subset in Neutrophil Recruitment in Chemically Induced Rat Pulmonary Inflammation.

There is growing support for the notion that chronic inflammation contributes to lung tumorigenesis, but the molecular and cellular basis underlying the protumorigenic effects of inflammation remains to be explored. 3-Methylcholanthrene and diethylnitrosamine were intratracheally instilled into rats to induce multistep lung carcinogenesis, and the presence of pulmonary inflammation was observed in addition to precancerous lesions. By leveraging single-cell RNA sequencing, we sought to unravel the mechanism underlying the inflammatory process at a higher resolution. A total of 14 cell types were identified in chemically treated and control rats. Chemical intervention introduced heterogeneity in cell type composition and gene expression patterns. Nonimmune cells were found to be the most affected, and two subpopulations of endothelial cells with diverse roles were defined. Car4-high endothelial cells were mainly responsible for angiogenesis, whereas Car4-low endothelial cells were involved in neutrophil recruitment, and adhesion between Car4-low endothelial cells and neutrophils was verified in inflamed tissues. Our work unveiled the intricate process of pulmonary inflammation at the single-cell level and characterized a proinflammatory subpopulation of endothelial cells involved in neutrophil recruitment. The conditions provided by chronic inflammatory environment are prerequisites for neoplastic progression. Targeting the specific subsets or processes defined herein holds promise for the early prevention and therapeutic intervention of lung cancer through the manipulation of angiogenesis or the inflammatory response.