calpain and Colorectal-Neoplasms

Myristoylation refers to the co-translational addition of a myristoyl group to an amino-terminal glycine residue of a protein by an ubiquitously distributed enzyme myristoyl-CoA:protein N-myristoyltransferase (NMT, EC 2.3.1.97). This review describes the basic enzymology, molecular cloning and regulation of NMT activity in various pathophysiological processes such as colon cancer and diabetes.

Topics: Acyl Coenzyme A; Acyltransferases; Adenocarcinoma; Animals; Calcium; Calpain; Colorectal Neoplasms; Enzyme Precursors; Gene Expression Regulation, Enzymologic; Humans; Kinetics; Molecular Sequence Data; Oncogene Protein pp60(v-src); Protein Processing, Post-Translational

Topics: ATP Binding Cassette Transporter 1; Calpain; Colorectal Neoplasms; Frameshift Mutation; Humans

Several studies have demonstrated that calpain‑1 is involved in a variety of pathophysiological processes, including tumorigenesis. However, the clinical relevance and role of calpain‑1 in colorectal cancer (CRC) are unclear. Filamin A (FLNA) is an actin‑binding protein that participates in cancer progression and can be cleaved by calpain‑1. In the present study, the protein expression levels of calpain‑1 and FLNA were detected by immunohistochemistry in 467 matched cancerous and paracancerous tissues from patients with CRC. The staining results and the clinicopathological characteristics of the patients were comprehensively analyzed. A high expression level of calpain‑1 was strongly associated with age, metastasis, Dukes stage and survival time but not with sex, histologic grade, tumour location or tumor size. By contrast, a low expression level of FLNA was significantly associated with tumor size, histological grade, metastasis, Dukes stage and survival time, but not with age, sex, or tumor location. Kaplan‑Meier survival analysis demonstrated that patients with calpain‑1 overexpression had a shorter mean overall survival (OS) than patients with lower levels of calpain‑1 expression. Unlike high levels of calpain‑1, high levels of FLNA were associated with longer OS than lower levels of FLNA expression. Furthermore, calpain‑1 expression was inversely correlated with FLNA expression. The relationship between calpain‑1 and FLNA was further confirmed using CRC cell lines in vitro. When calpain‑1 expression decreased in CRC cells, FLNA expression increased. Furthermore, calpain‑1 knockdown in CRC cells resulted in decreased proliferation, colony formation, migration and invasion. The present findings suggest that calpain‑1 overexpression predicted a poor outcome in patients with CRC and promoted tumor progression, possibly via FLNA downregulation.

Topics: Aged; Biomarkers, Tumor; Calpain; Cell Line, Tumor; Cell Movement; Cell Proliferation; Colorectal Neoplasms; Disease Progression; Female; Filamins; Gene Expression Regulation, Neoplastic; Humans; Kaplan-Meier Estimate; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Invasiveness

Accumulating evidence has suggested that calpain small subunit 1 (Capn4) plays an important role in the development and progression of malignant tumors. However, little is known about the role of Capn4 in colorectal cancer (CRC). In this study, we aimed to investigate the potential role of Capn4 in CRC and the regulation of Capn4 by microRNAs (miRNAs). Here, we found that Capn4 expression was highly up-regulated in CRC cell lines. Knockdown of Capn4 by siRNA significantly inhibited the proliferation and invasion of CRC cell lines. Furthermore, knockdown of Capn4 suppressed Wnt signaling in CRC cells. Interestingly, Capn4 was found to be a target gene of miR-1271, a tumor suppressive miRNA. The results showed that miR-1271 negatively regulated Capn4 expression in CRC cells. An inverse correlation between miR-1271 and Capn4 was also shown in CRC clinical tissues. Moreover, the overexpression of miR-1271 suppressed the proliferation, invasion and Wnt signaling of CRC cells. Importantly, we found that the restoration of Capn4 expression significantly reversed the antitumor effects of miR-1271 in CRC cells. Overall, these results suggest that miR-1271 inhibits the proliferation and invasion of CRC cells by down-regulating Capn4. Our study suggests that Capn4 and miR-1271 may serve as potential therapeutic targets for the treatment of CRC.

Topics: Base Sequence; Calpain; Cell Line, Tumor; Cell Proliferation; Colorectal Neoplasms; Down-Regulation; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Humans; MicroRNAs; Models, Biological; Neoplasm Invasiveness; Up-Regulation

Cancer-related cachexia involves increased protein breakdown through various proteolytic pathways, including the ubiquitin-proteasome pathway (UPP). We hypothesized that a calcium- and calpain-dependent pathway might play a crucial role during the proteolytic procedure, and that pathway interventions would ameliorate cancer cachexia in vivo. After being inoculated with CT26 adenocarcinoma cell culture subcutaneously, BALB/c mice developed cachexia in 12 days. They were then administered with different types of calpain inhibitors individually or in combination for 7 consecutive days. Eighteen healthy mice were also assessed as a control group. Changes in body weight, gastrocnemius muscle mass, tumor volume, food intake, survival time, and serum nutritional markers were monitored. Also measured were the levels of calpains, E3 ubiquitin ligases, and apoptosis-associated markers in gastrocnemius muscle. Our study showed that the intraperitoneal administration of calpain inhibitors significantly improved tumor-free body weight and gastrocnemius muscle mass in all treatment groups. Treatment with calpain inhibitors also ameliorated cachexia-associated negative effects in metabolic profiles and increased survival time in most of the tumor-bearing mice compared with the cachexia controls. Furthermore, calpain inhibitors reduced the calpain activity and the expression of MuRF-1 and atrogin-1 in all treatment groups, while increasing the level of cleaved caspase-3 and BAX and lowering the level of BCL-2 in some groups. These results justify further evaluation of calpain inhibitors both alone and in combination with other candidate agents as a potential new therapeutic strategy for treating cancer cachexia.

Topics: Adenocarcinoma; Animals; Apoptosis; Blotting, Western; Cachexia; Calpain; Colorectal Neoplasms; Disease Models, Animal; Glycoproteins; Humans; Male; Mice; Mice, Inbred BALB C; Proteolysis; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

CTEN/TNS4 is a member of the Tensin gene family. It localizes to focal adhesions and induces cell motility. The mechanisms regulating Cten expression are unclear although we have shown regulation by Kras in the colon and pancreas. In normal mammary cell lines, it is reportedly upregulated by epidermal growth factor receptor (EGFR) and STAT3 signalling and upregulation is accompanied by downregulation of Tensin 3 (Tensin switch). In this study, we investigated the roles of EGFR and STAT3 signalling in the regulation of Cten in colorectal cancer (CRC). In addition, we investigated calpain--a regulator of focal adhesion-associated proteins whose relevance to Cten has not been investigated. CRC cell lines were stimulated with epidermal growth factor (EGF). This resulted in an increase in Cten and Tensin 3 protein. Kras was knocked down and this resulted in downregulation of Cten and Tensin 3. We next investigated the role of STAT3 signalling. Activation and knockdown of STAT3 resulted in downregulation and upregulation, respectively, of Cten. Inhibition of calpain resulted in upregulation of both Cten and Tensin 3. As the regulators of Cten also seemed to regulate Tensin 3, we tested the interaction between Cten and Tensin 3. Cten was forcibly expressed or knocked down resulting, respectively, in upregulation and downregulation of Tensin 3. We conclude that in CRC, Cten is upregulated by EGFR and Kras but downregulated by STAT3. We show that calpain may be a negative regulator of Cten and that a Tensin switch does not occur and, if anything, Cten stabilizes Tensin 3.

Topics: Calpain; Colorectal Neoplasms; Cysteine Proteinase Inhibitors; Dipeptides; Dose-Response Relationship, Drug; Epidermal Growth Factor; ErbB Receptors; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; HCT116 Cells; Humans; Microfilament Proteins; Protein Stability; Proto-Oncogene Proteins p21(ras); Signal Transduction; STAT3 Transcription Factor; Tensins; Transfection

Insulin resistance (IR) is an established risk factor for colorectal cancer (CRC). Given that CRC and IR physiologically overlap and the calpain-10 gene (CAPN10) is a candidate for IR, we explored the association between CAPN10 and CRC risk.. Blood samples of 400 case-control pairs were genotyped, and the lifestyle and dietary habits of these pairs were recorded and collected. Unconditional logistic regression (LR) was used to assess the effects of CAPN10 SNP43 and SNP19, and environmental factors. Both generalized multifactor dimensionality reduction (GMDR) and the classification and regression tree (CART) were used to test gene-environment interactions for CRC risk.. The GA+AA genotype of SNP43 and the Del/Ins+Ins/Ins genotype of SNP19 were marginally related to CRC risk (GA+AA: OR = 1.35, 95% CI = 0.92-1.99; Del/Ins+Ins/ Ins: OR = 1.31, 95% CI = 0.84-2.04). Notably, a high-order interaction was consistently identified by GMDR and CART analyses. In GMDR, the four-factor interaction model of SNP43, SNP19, red meat consumption, and smoked meat consumption was the best model, with a maximum cross-validation consistency of 10/10 and testing balance accuracy of 0.61 (P < 0.01). In LR, subjects with high red and smoked meat consumption and two risk genotypes had a 6.17-fold CRC risk (95% CI = 2.44-15.6) relative to that of subjects with low red and smoked meat consumption and null risk genotypes. In CART, individuals with high smoked and red meat consumption, SNP19 Del/Ins+Ins/Ins, and SNP43 GA+AA had higher CRC risk (OR = 4.56, 95%CI = 1.94-10.75) than those with low smoked and red meat consumption.. Though the single loci of CAPN10 SNP43 and SNP19 are not enough to significantly increase the CRC susceptibility, the combination of SNP43, SNP19, red meat consumption, and smoked meat consumption is associated with elevated risk.

Topics: Adult; Aged; Aged, 80 and over; Calpain; Case-Control Studies; Colorectal Neoplasms; DNA; Feeding Behavior; Female; Gene-Environment Interaction; Genotype; Humans; Male; Meat; Middle Aged; Polymerase Chain Reaction; Polymorphism, Single Nucleotide; Prognosis; Risk Factors

Several studies have reported that plant-derived natural products have cancer chemopreventive and chemotherapeutic properties. The aim of the present study was to determine the antiproliferative and pro-apoptotic potential of Limoniastrum guyonianum aqueous gall extract (G extract) on human colorectal cancer BE cell line and, if so, to characterize the mechanism involved. The G extract-induced growth inhibitory effect was associated with an arrest of cell cycle progression in G2/M phase as shown by the cell phase distribution. In addition, G extract promoted in a concentration-dependent manner these cells towards apoptosis as indicated by the presence of cleaved poly(ADP-ribose) polymerase (PARP). In order to characterize the mechanism involved in the antiproliferative and pro-apoptotic signaling pathway activated by G extract, calpain activity and the expression of the cell cycle inhibitor p16(INK4A) were determined. The present findings indicated that G extract exhibited significant inhibitory activity against calpain and caused a marked and concentration-dependent upregulation of p16(INK4A). These effects could be ascribed to the presence of condensed tannins and polyphenols such as epicatechin and epigallocatechin gallate in G extract.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Calpain; Cell Cycle; Cell Proliferation; Colorectal Neoplasms; Cyclin-Dependent Kinase Inhibitor p16; Dipeptides; Humans; Plant Extracts; Plumbaginaceae; Poly(ADP-ribose) Polymerases

In this study, we have investigated the effects of luteolin on colorectal cancer cells. Our results demonstrate that luteolin is able to induce cytotoxicity and cell cycle perturbation in a dose-dependent manner. By triggering poly(ADP-ribose) polymerase (PARP) cleavage, this molecule is able to induce the apoptosis of BE colorectal cancer cells. We have also studied the potential involvement of calpains in the proapoptotic effects of luteolin. Our data show that luteolin exhibits moderate inhibitory activity against calpain. Thus, treatment of these cells with both luteolin and the calpain inhibitor MDL 28170 causes an increase in the luteolin-induced apoptosis as proved by the enhancement of 89- and 26-kDa PARP fragments. This effect is concomitant with the downregulation of the DNA methyltransferase 1 (DNMT1) expression and the epigenetic integrator ubiquitin-like containing PHD Finger 1 (UHRF1). As a result, luteolin induces an upregulation of a tumor suppressor gene: p16(INK4A). This study further proposes that calpain might be involved in the epigenetic code inheritance by regulating the epigenetic integrator UHRF1. We conclude from these results that targeting calpain, UHRF1, and DNMT1 using luteolin could be an interesting way to prevent and/or treat colorectal cancers.

Topics: Apoptosis; Calpain; CCAAT-Enhancer-Binding Proteins; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; Colorectal Neoplasms; Cyclin-Dependent Kinase Inhibitor p16; Dipeptides; DNA (Cytosine-5-)-Methyltransferase 1; DNA (Cytosine-5-)-Methyltransferases; Down-Regulation; Epigenesis, Genetic; Gene Expression Regulation, Neoplastic; Humans; Luteolin; Poly(ADP-ribose) Polymerases; Ubiquitin-Protein Ligases; Up-Regulation

Chymotrypsin-like activity of proteasomes and total calpain activity were studied in patients with stage T2-4N0-3M0 gastric cancer and stage T2-4N0-2M0-1 colorectal cancer. Activities of proteasomes and calpains in gastric and colorectal cancer tissues were higher than in the corresponding normal tissues. Changes in activities of proteasomes and calpains were mutually related. The appearance of lymphogenic metastases in gastric cancer was associated with the increase in calpain activity. The progress of colorectal cancer and development of lymphogenic and hematogenic metastases were associated with elevated chymotrypsin-like activity of proteasomes.

Topics: Calpain; Chymotrypsin; Colorectal Neoplasms; Fluorometry; Humans; Lymphatic Metastasis; Proteasome Endopeptidase Complex; Statistics, Nonparametric; Stomach Neoplasms

CPT-11 (irinotecan), the first-line chemotherapy for advanced stage colorectal cancer, remains inactive in about half of patients (primary chemoresistance) and almost all initial responders develop secondary resistance after several courses of treatment (8 months on average). Nude mice bearing HT-29 colon cancer xenografts were treated with CPT-11 and/or an NF-κB inhibitor for two courses. We confirm that NF-κB inhibition potentiated CPT-11 anti-tumoural effect after the first course of treatment. However, tumours grew again at the end of the second course of treatment, generating resistant tumours. We observed an increase in the basal NF-κB activation in resistant tumours and in two resistant sublines, either obtained from resistant HT-29 tumours (HT-29R cells) or generated in vitro (RSN cells). The decrease of NF-κB activation in HT-29R and RSN cells by stable transfections with the super-repressor form of IκBα augmented their sensitivity to CPT-11. Comparing gene expression profiles of HT-29 and HT-29R cells, we identified the S100A10/Annexin A2 complex and calpain 2 as over-expressed potential NF-κB inducers. SiRNA silencing of calpain 2 but not of S100A10 and/or annexin A2, resulted in a decrease in NF-κB activation, an increase in cellular levels of IκBα and a partial restoration of the CPT-11 sensitivity in both HT-29R and RSN cells, suggesting that calpain 2-dependent IκBα degradation mediates CPT-11 secondary resistance. Thus, targeted therapies directed against calpain 2 may represent a novel strategy to enhance the anti-cancer efficacy of CPT-11.

Topics: Animals; Annexin A2; Antineoplastic Agents; Apoptosis; Calpain; Camptothecin; Cell Proliferation; Colorectal Neoplasms; Drug Resistance, Neoplasm; Female; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; I-kappa B Proteins; Irinotecan; Mice; Mice, Inbred Strains; Mice, Nude; Neoplasm Transplantation; NF-kappa B; Pyrimidines; S100 Proteins; Transfection; Xenograft Model Antitumor Assays

Non-steroidal anti-inflammatory drugs (NSAIDs) have become promising agents for the chemoprevention of colorectal cancer presumably by inducing apoptosis. However, the role of NSAIDs in the regulation of NHE-1, intracellular calcium [Ca(2+)](i) homeostasis and Calpain 9 signaling pathway seems unclear. Male Sprague-Dawley rats were used as model for experimental colorectal cancer. Effects of pro-carcinogen (DMH) and NSAIDs were studied via morphological examination of tumors, histopathological changes, [Ca(2+)](i) homeostasis, production of reactive oxygen species (ROS), changes in mitochondrial membrane potential (MMP or ΔΨ(M)) and expression profile of target genes and proteins both at transcription and translation levels respectively. Size and number of tumors were found less in NSAIDs co-administered groups as compared to the DMH alone. Higher expression of NHE-1 was observed in DMH group whereas Calpain 9 was up-regulated in NSAIDs co-administered groups. [Ca(2+)](i) levels and reactive oxygen species were observed elevated in NSAIDs co-administered groups. ΔΨ(M) was found higher in DMH alone group along with the increased expression of the anti-apoptotic and mitochondrial membrane guard protein, Bcl-2. Expression levels of various pro-apoptotic proteins were observed higher in case of NSAIDs co-administered groups. Down-regulation of NHE-1, along with an increased [Ca(2+)](i) and induction of intrinsic pathway of apoptosis via activated Calpain 9 could be a putative mechanism pursued by Sulindac and Celecoxib for the chemoprevention of colorectal cancer.

Topics: 1,2-Dimethylhydrazine; Animals; Anti-Inflammatory Agents, Non-Steroidal; Apoptosis; Calcium; Calpain; Celecoxib; Colorectal Neoplasms; Down-Regulation; Gene Expression Regulation, Neoplastic; Male; Membrane Potential, Mitochondrial; Pyrazoles; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Sodium-Hydrogen Exchangers; Sulfonamides; Sulindac; Up-Regulation

Colorectal cancer (CRC) is one of the most prevalent types of cancer affecting both men and women in developed countries. Clinical and molecular evidence suggests that there are multiple biochemical pathways involved in its susceptibility, pathogenesis and prognosis. Several studies have reported a significant association between the incidence of CRC and type 2 diabetes mellitus (T2DM). However, genes associated with both conditions are rare.. We have analyzed the CAPN10 gene, a T2DM locus, using UCSNP-43, -44, -19, and -63 markers, looking for differences between 371 CRC patients and 605 unrelated controls of Spanish origin.. We found that UCSNP-44 allele C is swept out from cases (OR = 0.16, P = 0.005). Moreover, the frequency of 2111/2111 haplogenotype is also statistically lower than expected in CRC patients (P = 0.006).. Taken together, our results indicate a recessive model for the effect of CAPN10 variant UCSNP-44 influencing the risk of CRC and suggest a novel genetic link between T2DM and colon carcinoma.

Topics: Calpain; Case-Control Studies; Colorectal Neoplasms; Female; Gene Frequency; Genetic Markers; Genetic Predisposition to Disease; Haplotypes; Humans; Male

Calpains represent a well-conserved family of Ca2+ -dependent proteolytic enzymes. Recently, the importance of calpain in the metastatic process has received great attention. To investigate whether m-calpain contributes to the pathogenesis of colorectal cancer, we investigated the expression of m-calpain and its inhibitors, calpastatin and high-molecular-weight calmodulin-binding protein (HMWCaMBP), in human colorectal surgical specimens.. Fifty cases of colon carcinoma were evaluated for this study. Of 50 cases evaluated, we presented in this report six cases for m-calpain, calpastatin and HMWCaMBP protein expression by Western blot analyses was done in both normal and invasive tumor components of human samples. In addition, immunohistochemistry analysis was also carried out in all patients.. The activity and protein expression of m-calpain was significantly higher in colorectal adenocarcinoma than in normal colonic mucosa. This finding was corroborated by immunohistochemical studies that showed strong cytoplasmic staining in the colon tumors with m-calpain antibody. The decreased expression of these calpain inhibitors (calpastatin and HMWCaMBP) paralleled increased activity and expression of calpain in colorectal adenocarcinoma and the well-documented involvement of this Ca2+ -dependent protease in colon tumor.. Increased activity and moderate staining of m-calpain in polyps show the usage of this enzyme as a marker for the early detection of colorectal adenocarcinoma using immunologic approaches. These findings represent the first description of calpain overexpression in colorectal cancer. This has implications with regard to the design of chemotherapeutic drugs as well as in monitoring colorectal cancer in early stages of the metastatic process.

Topics: Adenocarcinoma; Biomarkers, Tumor; Calcium-Binding Proteins; Calmodulin-Binding Proteins; Calpain; Colorectal Neoplasms; Cysteine Proteinase Inhibitors; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry