Spermine oxidase targets thioredoxin reductase 1 to promote malignant progression of colon cancer

Objective: Epidermal growth factor (EGF) has many biological functions and plays an important role in the progression of various tumors including gastric cancer. An A-G single nucleotide polymorphism (SNP) at position 61 in the 5′-untranslated region (UTR) of the EGF gene has recently been reported to be associated with different levels of EGF production. We examined whether this polymorphism is correlated with the development and malignant phenotypes of gastric cancer. Methods: The study population included 200 gastric cancer patients and 230 healthy control subjects. The SNP in the 5′-UTR of the EGF gene was analyzed by polymerase chain reaction-restriction fragment length polymorphism. Results: The A allele was significantly less frequent in patients than in controls (p = 0.01). Individuals with the A/A or A/G genotype showed a significantly lower risk of gastric cancer than those with the G/G genotype [adjusted odds ratio (OR) = 0.56], whereas the same genotypes were associated with malignant progression of this cancer, e.g. deeper tumor invasion, increased lymph node metastasis and advanced clinical stage, and histological classification in gastric cancer patients (adjusted OR = 1.80, 1.98, 2.26 and 1.89, respectively). Conclusions: Our findings suggest that the A-G polymorphism of EGF is involved not only in the occurrence but also in the malignant progression of gastric cancer.

DCLK1 and its oncogenic functions: A promising therapeutic target for cancers

BackgroundThe tumor microenvironment (TME) is not only a key factor in the malignant progression of cancer but also plays an indispensable role in tumor immunotherapy. As an important regulatory factor in the TME, long non-coding RNAs (incRNA) are important for the development of bladder cancer. The purpose of this study was to explore the molecular mechanism of malignant progression of bladder cancer (BCa) from the perspective of immunology, establish a reliable signature, and evaluate its effect on prognosis, metastasis, and the effectiveness of immunotherapy.MethodsThe TME was assessed by single-sample gene set enrichment analysis (ssGSEA) in 373 patients with muscle invasive bladder cancer (MIBC) in The Cancer Genome Atlas (TCGA). Combining RNA sequence data from 49 BCa patients in our center, we established TME-related prognostic signatures (TMERPS) based on TME-related immune prognosis genes using weighted gene correlation network analysis, selection operator Cox analysis, minimum absolute shrinkage, and survival analysis. Real-Time Quantitative PCR was used for expression level analysis of related genes. Functional enrichment analysis and nomograms were used to explore the potential impact of TMERPS on the immune system, prognosis, and metastasis.ResultsThe ssGSEA proved to be an accurate assessment of immune levels in BCa samples. TMERPS was established based on six TME-associated prognostic lncRNAs and was shown to be closely associated with prognosis, metastasis, and immune levels, and to have a significant stratifying effect on the therapeutic efficacy of immune checkpoint inhibitors. Finally, three TMERPS-based nomograms were shown to be effective in predicting prognosis, lymph node metastasis, and distant metastasis in BCa patients.ConclusionsTMERPS can stratify BCa patients into different risk groups with different prognoses, immunotherapy sensitivity, and risk of metastasis. TMERPS-based nomograms can effectively predict prognosis and metastasis in BCa patients.

This study aims to reveal the potential effect of circNBPF10 on the malignant progression of lung cancer. The expression levels of circNBPF10 in lung cancer tissues and cell lines were detected via real-time quantitative PCR (RT-qPCR). The relationship between circNBPF10 expression and lung cancer metastasis was further analyzed. Effects on lung cancer cells after the knockout or overexpression of circNBPF10 were detected. Subsequently, the regulatory relationship of circNBPF10 with miR-224 was detected by using the dual-luciferase reporter gene. In addition, the role of pre-B-cell homeo box 3 (PBX3) in the progression of lung cancer affected by circNBPF10 was evaluated through a rescue experiment. circNBPF10 was highly expressed in lung cancer tissues and lung cancer cell lines. The expression level of circNBPF10 was significantly higher in patients with lung cancer and lymphatic metastasis or distant metastasis than in patients with nonmetastatic lung cancer. The downregulation of circNBPF10 reduced the proliferation, migration, and invasion of lung cancer cells. In lung cancer cells, circNBPF10 negatively regulated the expression of miR-224, whereas miR-224 directly targeted the expression of PBX3. The results of the rescue experiment confirmed that PBX3 was the key gene for the promoting effect of circNBPF10 on the malignant progression of lung cancer. circNBPF10 was highly expressed in lung cancer tissues and was associated with distant metastasis and poor prognosis in patients with lung cancer. circNBPF10 upregulated PBX3 by targeting miR-224 and promoted the malignant progression of lung cancer.

Obesity and diet‐induced overweight have been demonstrated as risk factors of colon cancer. Several studies suggested that loss of E‐cadherin adherent molecule has been associated with epithelial– mesenchymal transition (EMT) and the malignant progression of human colon cancer cells. However, the effects of high fat (HF) diet intake on the expression of E‐cadherin and in vivo malignant progression of human colon cancer have not been demonstrated well yet. Our results indicated that consumption of HF diet (45% of total energy intake) for 4 weeks would induce the tumor growth in human colon cancer HT‐29 cell bearing mice. Bioluminescence imaging analysis also confirmed that consumption of HF diet would stimulate the proliferation of human colon cancer. HF diet modulated the expression of E‐cadherin and β‐catenin in xenograft mouse model. Furthermore, consumption of HF diet enhanced the cellular proliferation through the modulation of proliferating cellular nuclear antigen (PCNA) and p21cip proteins. The possible molecular mechanisms of actions were, in part, through the activation of the MAPK/ERK signaling pathway and the suppression of E‐cadherin adhesion molecule. Take together; these results suggested that consumption of HF diet could modulate the growth and the malignant progression of human colon cancer cells in vivo.

BackgroundExtracellular matrix (ECM) is a mediator of tumor progression. However, whether the alterations of the intraperitoneal ECM prior to tumor establishment affects the malignant progression of ovarian cancer remains elusive.MethodsApolipoprotein (ApoE) knock-out mice was used to analyze the intraperitoneal ECM alterations by quantification of the major components of ECM. ID8 cells were implanted in vivo to generate allografts and human ovarian cancer cell lines were characterized in vitro to assess the effects of ECM alterations on the malignant progression of ovarian cancer. Adhesion assay, immunochemistry, cytokines profile, proliferation assay, transwell invasion assay and western blot were used to determine the malignant phenotype of ovarian cancer cells.ResultsApoE loss induced increased ECM deposition, which stimulated the adhesions of ovarian cancer cells. The adhesion-mediated focal adhesion kinase (FAK) signaling enhanced the invasive behaviors of ovarian cancer cells through activation of a ERK-MMP linkage. This ECM-induced signaling cascade was further confirmed in human ovarian cancer cell lines in vitro. Furthermore, reversal of the ECM accumulation with BAPN or abrogation of adhesion-induced ERK activation in ovarian cancer cells with MEK inhibitors (MEKi) was found to effectively delay ovarian cancer progression.ConclusionsThese findings identify the FAK-ERK activation in cell/matrix adhesion in the malignant progression of ovarian cancer and the efficiency of BAPN or MEKi for tumor suppression, providing an impetus for further studies to explore the possibility of new anticancer therapeutic combinations.

Recently, circular RNA was reported to be a significant participant in the development of tumorigenesis, including colorectal cancer. Therefore, we aimed to clarify the precise role of circ-keratin 6C (circ-KRT6C) in colorectal cancer progression. The relative expression levels of circ-KRT6C, microRNA-485-3p (miR-485-3p), and programmed cell death receptor ligand 1 (PDL1) were analyzed by real-time quantitative polymerase chain reaction and Western blot assays. The proliferation was assessed by cell count kit 8 and colony-forming assays. The apoptotic cells were determined by flow cytometry assay. The migration and invasion were analyzed by transwell assay. Colorectal cancer cells were cocultured with peripheral blood mononuclear cells or cytokine-induced killer cells to assess immune response. The interaction relationships among circ-KRT6C, miR-485-3p, and PDL1 were examined by dual-luciferase reporter assay. The effects of circ-KRT6C inhibition in vivo were analyzed by an animal experiment. circ-KRT6C was overexpressed in colorectal cancer tissues and cells, and its level was associated with overall survival time of patients with colorectal cancer. The suppression of circ-KRT6C suppressed growth, migration, invasion, and immune escape while stimulating apoptosis in colorectal cancer cells, which was abolished by shortage of miR-485-3p. In addition, overexpression of miR-485-3p repressed malignant progression and immune evasion of colorectal cancer by targeting PDL1, implying that PDL1 was a functional target of miR-485-3p. A xenograft experiment also suggested that circ-KRT6C inhibition could repress tumor growth in vivo. circ-KRT6C could increase PDL1 expression by functioning as an miR-485-3p sponge, which promoted malignant progression and immune evasion of colorectal cancer cells. SIGNIFICANCE STATEMENT: circ-keratin 6c could increase programmed cell death receptor ligand 1 expression by functioning as a microRNA-16-5p sponge, which promoted malignant progression and immune evasion of colorectal cancer.

Isolation and characterization of the major form of human MUC18 cDNA gene and correlation of MUC18 over-expression in prostate cancer cell lines and tissues with malignant progression

Solar ultraviolet (UV) radiation can cause severe damage to the skin and is the primary cause of most skin cancer. UV radiation causes DNA damage leading to mutations and also activates the Erbb2/HER2 receptor through indirect mechanisms involving reactive oxygen species. We hypothesized that Erbb2 activation accelerates the malignant progression of UV-induced skin cancer. Following the induction of benign squamous papillomas by UV exposure of v-ras(Ha) transgenic Tg.AC mice, mice were treated topically with the Erbb2 inhibitor AG825 and tumor progression monitored. AG825 treatment reduced tumor volume, increased tumor regression, and delayed the development of malignant squamous cell carcinoma (SCC). Progression to malignancy was associated with increased Erbb2 and ADAM12 (A Disintegin And Metalloproteinase 12) transcripts and protein, while inhibition of Erbb2 blocked the increase in ADAM12 message upon malignant progression. Similarly, human SCC and SCC cell lines had increased ADAM12 protein and transcripts when compared to normal controls. To determine whether Erbb2 up-regulation of ADAM12 contributed to malignant progression of skin cancer, Erbb2 expression was modulated in cultured SCC cells using forced over-expression or siRNA targeting, demonstrating up-regulation of ADAM12 by Erbb2. Furthermore, ADAM12 transfection or siRNA targeting revealed that ADAM12 increased both the migration and invasion of cutaneous SCC cells. Collectively, these results suggest Erbb2 up-regulation of ADAM12 as a novel mechanism contributing to the malignant progression of UV-induced skin cancer. Inhibition of Erbb2/HER2 reduced tumor burden, increased tumor regression, and delayed the progression of benign skin tumors to malignant SCC in UV-exposed mice. Inhibition of Erbb2 suppressed the increase in metalloproteinase ADAM12 expression in skin tumors, which in turn increased migration and tumor cell invasiveness.

As a member of the DUBs (deubiquitinating enzymes) family, USPs (ubiquitin-specific peptidases) play a crucial regulatory role in the occurrence and progression of various malignancies. This review summarizes the current research on USPs in colorectal cancer, with a focus on the involvement of different USPs as either oncogenes or tumor suppressor genes in regulating the malignant progression of colorectal cancer. We discuss in detail the regulatory role of USPs in the process of drug resistance in tumors, the signaling pathways regulated by USPs, the correlation between different USP expression levels and early diagnosis and prognostic evaluation, as well as the latest research developments in USP inhibitors. Mechanistically, USPs regulate key proteins in several signaling pathways, including Wnt, P53, and PI3K/Akt/mTOR, highlighting their significant role in the malignant progression of colorectal cancer. Finally, we further discuss the functional and mechanistic differences among various USPs, the challenges faced in developing USP inhibitors, and prospects for future research directions.

METCAM/MUC18, an integral membrane cell adhesion molecule (CAM) in the Ig-like gene superfamily, is capable of performing typical functions of CAMs, such as cell-cell and cell-extracellular interactions, crosstalk with intracellular signaling pathways, and modulating social behaviors. METCAM/MUC18 is not expressed in >90% of the epithelial cells of normal prostate, or in 100% of benign prostatic hyperplasia (BPH), but is expressed in >80% of prostatic intracellular neoplasia (PIN), high grade prostate cancers, and metastatic lesions. Its expression is also correlated with the malignant progression of mouse prostate adenocarcinoma in a transgenic model, TRAMP. Overexpression of human METCAM/MUC18 increases epithelial-to-mesenchymal transition (in vitro motility and in vitro invasiveness) of prostate cancer cells and in vivo tumorigenesis and metastasis to multiple organs after orthotopic injection of human prostatic cancer LNCaP cells in male nude mice. From our preliminary studies, it appears to regulate these processes via increasing proliferation, up-regulating the AKT-signaling pathway, increasing aerobic glycolysis, and augmenting angiogenesis of prostate cancer cells, but has no effect on apoptosis. Furthermore, soluble METCAM/MUC18 could block angiogenesis of LNCaP tumors and specific shRNAs in a lentivirus vector block tumorigenesis of DU145 cells in an athymic nude mouse model. Taken together, METCAM/MUC18 may be a useful novel biomarker for early diagnosis of the malignant potential of prostate cancer, but also a metastatic progression gene to drive the malignant progression of prostate cancer in a pre-clinic mouse model. METCAM/ MUC18-specific siRNAs and its derived oligo-peptides may be useful as therapeutic agents to block the malignant progression of the cancer.

The Role of Type IV Collagen and Basement Membranes in Cancer Progression and Metastasis

Long noncoding RNAs (lncRNAs) regulate cancer progression and drug resistance. However, the role of lncRNA FGD5-AS1 in regulating colon cancer (CC) progression is still largely unknown. Hence, this study investigated the role of lncRNA FGD5-AS1 in regulating colon cancer (CC) progression and found that lncRNA FGD5-AS1 regulated miR-497-5p/PD-L1 axis to promote cancer progression in CC cells in vitro and in vivo. Specifically, we found that lncRNA FGD5-AS1 and PD-L1 tended to be high-expressed, while miR-497-5p was low-expressed in CC tissues and cell lines compared to the normal adjacent tissues and cells. Next, we found that lncRNA FGD5-AS1 positively regulated PD-L1 in CC cells by sponging miR-497-5p. Finally, our gain- and loss-of-function experiments evidenced that the lncRNA FGD5-AS1/miR-497-5p/PD-L1 axis regulates CC progression. Functionally, the data suggested that lncRNA FGD5-AS1 positively regulated while miR-497-5p negatively modulated malignant phenotypes, including cell proliferation, viability, invasion, migration, epithelial-mesenchymal transition (EMT), and tumorigenesis in CC cells. Interestingly, the inhibiting effects of lncRNA FGD5-AS1 ablation on CC development were abrogated by both silencing miR-497-5p and upregulating PD-L1. This study found that lncRNA FGD5-AS1 sponged miR-497-5p to upregulate PD-L1, resulting in CC progression, and provided novel agents for CC diagnosis and prognosis.

Introduction: Fusion between tumor cells with leukocytes contributes to tumor heterogeneity, resulting in tumorigenesis, metastasis, and treatment resistance. Recent studies demonstrate that an inflammatory microenvironment enhances tumor-macrophage fusion. We hypothesize that tumor-macrophage fusion hybrids contribute to radiation induced pancreatic cancer progression. Materials and Methods: Tumor tissues of female patients receiving previous bone marrow transplantation (BMT) from male donors due to leukemia/lymphoma were collected for Y chromosome and immunofluorescence study. Murine subcutaneous tumor models of cre-loxP or dual fluorescence system were established to demonstrate fusion between Pan02, a murine pancreatic adenocarcinoma cell line, and stromal cells of ROSA (Gt(ROSA)26Sor tm4(ACTB-tdTomato,-EGFP)Luo/Jnarl) mice with or without local radiation exposure. KC (pdx-1-cre-KrasLSL-G12D) mice, were given whole body irradiation followed by BMT from ROSA mice to measure fusion between pancreatic cancer and hematopoietic cells. Liposomal chlodronate, a macrophage depleting agent, and immunofluorescence staining of fusion hybrids in ROSA mice tumors were used to demonstrate macrophage as a fusion partner. Fusion hybrid clones of murine/human pancreatic adenocarcinoma cells with macrophages were selected from co-culture and sorting followed by studies including ploidy, clonogenicity, migration and radio-sensitivity. Affymetrix microarray analysis was used to find differentially expressed genes of fusion hybrids. Results: Fusion phenomenon was demonstrated by showing colocalization of Y chromosomes with epithelial markers in cancer tissues from female patients receiving prior BMT from male donors. Fusion hybrids were found in pancreas tumors of KC mice after BMT from ROSA mice. Fusion hybrids within tumors increased by time, three folds within 6 weeks; and 2.5 folds after 20Gy radiation exposure. Liposomal clodronate reduced fusion hybrids and tumor growth. Immunofluorescence stain of tumor tissues demonstrated CD11b(+) macrophage to be the major fusion partner. Purified Fusion hybrids showed enhanced migration, proliferation, aneuploidy and radio-resistance. Model based gene analysis of microarray study disclosed signal regulatory protein (SIRP) signal pathway was significantly upregulated in fusion hybrids. Upregulated SIRP signal molecules, including DAP12, PyK2, were validated by immunoblots. Silencing DAP12 in myeloid cells from human peripheral blood showed decreased fusion rate with Panc-1 cells. DAP12mRNA depletion in Panc-1 fusion hybrids disclosed enhanced radio-sensitivity. Defactinib (VS6063), a PyK2 inhibitor, reduced fusion phenomenon and suppressed pancreatic tumor growth of mice after local radiation. Conclusions: Fusion between pancreatic cancer cells and macrophages contribute to radiation enhanced malignant progression of pancreatic cancer. SIRP signal pathway is a potential target to disrupt fusion phenomenon and ameliorate pancreatic cancer progression after radiation. Citation Format: Hui Ju Ch'ang, Yi-Chih Tsai, Ting Wei Li, Rui-Jun Liu, Tze-Sing Huang, Sse-Pei Wan, Su-Liang Chen, Shih-Sheng Jiang. Fusion between macrophages and cancer cells up-regulates signal regulatory protein pathway and contributes to malignant progression of pancreatic cancer after radiation [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr A030.

Bladder cancer ranks as the second most prevalent urology malignancy globally. Invasive metastasis is a significant contributor to mortality among patients with bladder cancer, yet the underlying mechanisms remain elusive. Deubiquitinases are pivotal in carcinogenesis, with USP5 implicated in the malignant progression of hepatocellular carcinoma, colorectal cancer and non-small cell lung cancer. The present study assessed the role and mechanism of ubiquitin-specific proteinase 5 (USP5) in the malignant progression of bladder cancer. The association between USP5 expression and bladder cancer prognosis and stage was analyzed using The Cancer Genome Atlas database. Moreover, to elucidate the role of USP5 in bladder cancer, USP5 overexpression and knockdown cell lines were established using T24 cells. Cell viability, proliferation and migration were assessed using Cell Counting Kit-8, Transwell and scratch assays, respectively. Cyclohexanamide was used to evaluate the effect of USP5 expression on Snail family zinc finger 2 (SLUG) stability. Immunoprecipitation and immunofluorescence co-localization were utilized to probe the interaction between USP5 and SLUG. Changes in mRNA and protein levels were assessed using reverse transcription-quantitative PCR and western blotting, respectively. The results revealed that patients with bladder cancer with high USP5 expression had significantly shorter survival (P<0.05) and a higher clinicopathologic stage (P<0.05) than those with low USP5 expression. T24 cells overexpressing USP5 demonstrated significantly increased proliferation (P<0.05), invasion (P<0.05) and expression of epithelial-mesenchymal transition markers (P<0.05); whereas T24 cells with knocked-down USP5 expression revealed significantly reduced proliferation (P<0.05), invasion (P<0.05) and epithelial-mesenchymal transition markers (P<0.05). Immunoprecipitation experiments demonstrated the binding of USP5 to SLUG in bladder cancer cells, with further analysis revealing that USP5 upregulated protein levels of SLUG by inhibiting its ubiquitination. Furthermore, the treatment of bladder cancer cells with Degrasyn, a USP5 inhibitor, was associated with a significant inhibition of the proliferation (P<0.05) and invasion (P<0.05) of T24 cells. In conclusion, the findings of the present study underscore the role of USP5 in promoting the malignant progression of bladder cancer through the stabilization of SLUG. Targeting USP5 holds promise as a strategy for inhibiting bladder cancer progression.