calpain and Carcinogenesis

Calpain-1 and calpain-2 are prototypical classical isoforms of the calpain family of calcium-activated cysteine proteases. Their substrate proteins participate in a wide range of cellular processes, including transcription, survival, proliferation, apoptosis, migration, and invasion. Dysregulated calpain activity has been implicated in tumorigenesis, suggesting that calpains may be promising therapeutic targets.. This review covers clinical and basic research studies implicating calpain-1 and calpain-2 expression and activity in tumorigenesis and metastasis. We highlight isoform specific functions and provide an overview of substrates and cancer-related signalling pathways affected by calpain-mediated proteolytic cleavage. We also discuss efforts to develop clinically relevant calpain specific inhibitors and spotlight the challenges facing inhibitor development.. Rationale for targeting calpain-1 and calpain-2 in cancer is supported by pre-clinical and clinical studies demonstrating that calpain inhibition has the potential to attenuate carcinogenesis and block metastasis of aggressive tumors. The wide range of substrates and cleavage products, paired with inconsistencies in model systems, underscores the need for more complete understanding of physiological substrates and how calpain cleavage alters their functions in cellular processes. The development of isoform specific calpain inhibitors remains an important goal with therapeutic potential in cancer and other diseases.

Topics: Apoptosis; Calpain; Carcinogenesis; Humans; Neoplasms; Protein Isoforms

A systematic search was conducted against several online databases. Hazard ratios (HRs) or odds ratio (ORs) were used to investigate the relationship between Capn4 protein expression and prognosis as well as clinical parameters in cancer survivors.. Eleven studies involving 1775 patients were identified. Overall, the results showed that Capn4 protein was associated with poor prognosis of overall survival (OS) (HR=1.74; 95% CI:1.47-2.01;. Expression of Capn4 protein is associated with cancer survival and clinicopathologic characteristics in patients.

Topics: Calpain; Cancer Survivors; Carcinogenesis; Databases, Factual; Disease-Free Survival; Female; Humans; Male; Odds Ratio; Prognosis

Sarcomas are still unsolved therapeutic challenges. Cancer stem cells are believed to contribute to sarcoma development, but lack of specific markers prevents their characterization and targeting. Here, we show that calpain-6 expression is associated with cancer stem cell features. In mouse models of bone sarcoma, calpain-6-expressing cells have unique tumor-initiating and metastatic capacities. Calpain-6 levels are especially high in tumors that have been successfully propagated in mouse to establish patient-derived xenografts. We found that calpain-6 levels are increased by hypoxia in vitro and calpain-6 is detected within hypoxic areas in tumors. Furthermore, calpain-6 expression depends on the stem cell transcription network that involves Oct4, Nanog, and Sox2 and is activated by hypoxia. Calpain-6 knockdown blocks tumor development in mouse and induces depletion of the cancer stem cell population. Data from transcriptomic analyses reveal that calpain-6 expression in sarcomas inversely correlates with senescence markers. Calpain-6 knockdown suppresses hypoxia-dependent prevention of senescence entry and also promotion of autophagic flux. Together, our results demonstrate that calpain-6 identifies sarcoma cells with stem-like properties and is a mediator of hypoxia to prevent senescence, promote autophagy, and maintain the tumor-initiating cell population. These findings open what we believe is a novel therapeutic avenue for targeting sarcoma stem cells.

Topics: Animals; Autophagy; Biomarkers; Calpain; Carcinogenesis; Cell Line, Tumor; Cellular Senescence; Disease Models, Animal; Gene Expression Profiling; Gene Knockdown Techniques; Humans; Hypoxia; Male; Mice; Mice, Inbred BALB C; Microtubule-Associated Proteins; Nanog Homeobox Protein; Neoplasms; Neoplastic Stem Cells; Octamer Transcription Factor-3; Sarcoma; SOXB1 Transcription Factors; Xenograft Model Antitumor Assays

Accumulating evidence reports that Capn4 plays an important role in the development and progression of various malignant cancers. However, whether Capn4 is involved in prostate cancer remains unclear. Therefore, the aim of this study was to investigate the expression, biological function and regulatory mechanism of Capn4 in prostate cancer. Herein, we found that Capn4 was highly expressed in prostate cancer cell lines compared with normal prostate cells. Capn4 gene silencing markedly suppressed the growth, invasion and Wnt/β-catenin signaling of prostate cancer cells, whereas Capn4 overexpression showed an oncogenic effect. Moreover, silencing of β-catenin significantly blocked the oncogenic effect of Capn4 overexpression. Bioinformatics analysis predicted that Capn4 was a potential target gene of microRNA-520b (miR-520b), which has been reported as a tumor suppressive miRNA in various cancers. The dual-luciferase reporter assay confirmed that miR-520b directly bound to the 3'-untranslated region of Capn4. Real-time quantitative PCR and Western blot analysis showed that miR-520b negatively regulated Capn4 expression in prostate cancer cells in vitro. Furthermore, we found that miR-520b was significantly downregulated in prostate cancer cell lines and tissues. In addition, miR-520b expression was inversely correlated with Capn4 expression in prostate cancer clinical specimens. Overexpression of miR-520b mimicked the tumor suppressive effect of Capn4 siRNA, whereas inhibition of miR-520b had an oncogenic effect. Restoration of Capn4 significantly blocked the antitumor effect of miR-520b in prostate cancer cells. Overall, our findings demonstrate an oncogenic role of Capn4 in prostate cancer and show that its expression is epigenetically regulated by miR-520b. Our results reveal that suppression of Capn4 by miR-520b inhibits the growth and invasion of prostate cancer cells associated with downregulated Wnt/β-catenin signaling, indicating an important role of the miR-520b/Capn4/Wnt/β-catenin regulation axis in the molecular pathogenesis of prostate cancer. Our study suggests that miR-520b and Capn4 may represent potential and novel therapeutic targets for prostate cancer.

Topics: 3' Untranslated Regions; beta Catenin; Calpain; Carcinogenesis; Cell Line; Cell Line, Tumor; Cell Movement; Cell Proliferation; Down-Regulation; Gene Expression Regulation, Neoplastic; Humans; Male; MicroRNAs; Oncogenes; Prostate; Prostatic Neoplasms; RNA, Small Interfering; Wnt Signaling Pathway

Renal medullary carcinoma (RMC) is a rare and highly aggressive neoplasm that most often occurs in the setting of sickle cell trait or sickle cell disease (SCD). Most patients present with metastatic disease resistant to conventional chemotherapy, and therefore there is an urgent need for molecular insight to propose new therapies.. To determine the molecular alterations and oncogenic pathways that drive RMC development.. A series of five frozen samples of patients with RMC was investigated by means of gene expression profiling, array comparative genomic hybridization, and RNA and whole exome sequencing (WES).. RNA and DNA sequencing read data were analyzed to detect gene fusions and somatic mutations. Gene fusions mutations were validated by real-time polymerase chain reaction and fluorescence in situ hybridization. Gene expression profiling was analyzed by unsupervised hierarchical clustering and Gene Set Enrichment Analysis (Broad Institute, Cambridge, MA, USA).. We observed inactivation of the tumor suppressor gene SMARCB1 in all tumors. In all four cases developed in patients with SCD, we identified an original mechanism of interchromosomal balanced translocations that disrupt the SMARCB1 sequence and thus contribute to its inactivation. Gene expression profiling revealed that RMC shares common oncogenic pathways with pediatric malignant rhabdoid tumors, another tumor subtype characterized by SMARCB1 deficiency.. RMCs are characterized by an original mechanism of interchromosomal balanced translocations that disrupt the SMARCB1 sequence. WES reveals that RMCs show no other recurrent genetic alteration and an overall stable genome, underscoring the oncogenic potency of SMARCB1 inactivation.. Our comprehensive molecular study supports a pivotal role of the tumor suppressor gene SMARCB1 in the development of renal medullary carcinoma. The use of therapeutic strategies based on the biologic effects of its inactivation should now open new perspectives for this typically lethal malignancy.

Topics: Adolescent; Adult; Anemia, Sickle Cell; Calpain; Carcinogenesis; Carcinoma; Child; Comparative Genomic Hybridization; DNA-Binding Proteins; Exome Sequencing; Gene Expression Profiling; Gene Fusion; Humans; Kidney Neoplasms; Nuclear Proteins; Nuclear Receptor Subfamily 1, Group F, Member 1; RNA, Long Noncoding; Sequence Analysis, RNA; SMARCB1 Protein; Trans-Activators; Transcription Factors; Translocation, Genetic

Macrophages highly populate tumour microenvironment and are referred to as tumor-associated macrophages (TAMs). The inflammasome is a multiprotein complex responsible of IL-1 like cytokines release, which biology has been widely studied by using bone-marrow-derived macrophages to mimic a physiological and/or host defense condition. To understand the role of this complex in lung tumor-associated macrophages (TAMs), we isolated and cultured broncho-alveolar lavage (BAL)-derived cells of lung tumor-bearing mice. The stimulation of lung TAMs with LPS+ATP increased the release of IL-1β. The inhibition of NLRP3 by means of glybenclamide significantly reduced IL-1β release. Similarly, C3H-derived, caspase-1 ko and caspase-11 ko TAMs released significantly reduced levels of IL-1β. Moreover, the stimulation of lung TAMs with the sole LPS induced a significant release of IL-1α, which was significantly reduced after caspase-1 pharmacological inhibition, and in TAMs genetically lacking caspase-1 and caspase-11. The inhibition of calpain I/II by means of MDL28170 did not alter IL-1α release after LPS treatment of lung TAMs. To note, the inoculation of LPS-treated bone marrow-derived macrophages into carcinogen-exposed mice increased lung tumor formation. In contrast, the depletion of TAMs by means of clodronate liposomes reduced lung tumorigenesis, associated to lower in vivo release of IL-1α and IL-1β.In conclusion, our data imply lung tumor lesions are populated by macrophages which pro-tumor activity is regulated by the activation of the NLRP3 inflammasome that leads to the release of IL-1α and IL-1β in a caspase-11/caspase-1-dependent manner.

Topics: Adenosine Triphosphate; Animals; Bronchoalveolar Lavage Fluid; Calpain; Carcinogenesis; Caspase 1; Caspase Inhibitors; Caspases; Caspases, Initiator; Cells, Cultured; Clodronic Acid; Dipeptides; Female; Glyburide; Inflammasomes; Interleukin-1alpha; Interleukin-1beta; Lipopolysaccharides; Lung Neoplasms; Macrophages, Alveolar; Methylnitrosourea; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Mice, Knockout; Neoplasms, Experimental; NLR Family, Pyrin Domain-Containing 3 Protein; Signal Transduction; Specific Pathogen-Free Organisms; Toll-Like Receptor 4

SerpinB3 has been recently described as an early marker of liver carcinogenesis, but the potential mechanistic role of this serpin in tumor development is still poorly understood. Overexpression of Myc often correlates with more aggressive tumour forms, supporting its involvement in carcinogenesis. Yes-associated protein (Yap), the main effector of the Hippo pathway, is a central regulator of proliferation and it has been found up-regulated in hepatocellular carcinomas. The study has been designed to investigate and characterize the interplay and functional modulation of Myc by SerpinB3 in liver cancer. Results from this study indicate that Myc was up-regulated by SerpinB3 through calpain and Hippo-dependent molecular mechanisms in transgenic mice and hepatoma cells overexpressing human SerpinB3, and also in human hepatocellular carcinomas. Human recombinant SerpinB3 was capable to inhibit the activity of Calpain in vitro, likely reducing its ability to cleave Myc in its non oncogenic Myc-nick cytoplasmic form. SerpinB3 indirectly increased the transcription of Myc through the induction of Yap pathway. These findings provide for the first time evidence that SerpinB3 can improve the production of Myc through direct and indirect mechanisms that include the inhibition of generation of its cytoplasmic form and the activation of Yap pathway.

Topics: Adaptor Proteins, Signal Transducing; Animals; Antigens, Neoplasm; Calpain; Carcinogenesis; Carcinoma, Hepatocellular; Gene Expression Regulation, Neoplastic; Hep G2 Cells; Humans; Liver Neoplasms; Mice; Mice, Transgenic; Phosphoproteins; Proto-Oncogene Proteins c-myc; Serpins; Transcription Factors; Transcription, Genetic; YAP-Signaling Proteins

The colon cancer tissues from DMH treated rats exhibited higher membrane potential, fluidity and changed lipid order as examined by Merocyanine 540 and 1,6-diphenyl-1,3,5-hexatriene, respectively. A transition from gel to liquid crystalline state was observed by Laurdan fluorescence and also reduced fluorescence quenching of NBD-PE as contributed in the decreased membrane lipid phase separation. With piroxicam, a traditional NSAID and c-phycocyanin, a biliprotein from Spirulina platensis, these effects were normalized. An augmented intracellular Ca(+2) had contributed to the drug mediated apoptosis which is supported by an elevated calpain-9 expression. Histopathologically, a large pool of secreted acid/neutral mucopolysaccrides as well as the presence of blood vessels and dysplastic crypts signifies invasive mucinous adenocarcinoma while both the drugs reduced these neoplastic alterations. Wnt/β-catenin pathway was also found to be up-regulated which served as a crucial indicator for cancer cell growth. A concomitant down regulation of PPARγ was noted in DMH treatment which is associated with tumor progression. The expression of PPARα and δ, the other two isoforms of PPAR family was also modulated. We conclude that piroxicam and c-phycocyanin exert their anti-neoplastic effects via regulating membrane properties, raising calpain-9 and PPARγ expression while suppressing Wnt/β-catenin signaling in experimental colon carcinogenesis.

Topics: 2-Naphthylamine; Animals; Apoptosis; Calcium; Calpain; Carcinogenesis; Colonic Neoplasms; Fluorescence Polarization; Fura-2; Intracellular Space; Laurates; Ligands; Male; Membrane Fluidity; Membrane Potentials; Phase Transition; Phosphatidylethanolamines; Phycocyanin; Piroxicam; PPAR gamma; Pyrimidinones; Rats, Sprague-Dawley; Up-Regulation; Wnt Signaling Pathway