Dataset Information

The potential roles of a laminin receptor in adhesion and apoptosis of cells of the marine bivalve Meretrix meretrix.

ABSTRACT:

Background

The laminin receptors (LRs) play important roles in cell adhesion to the extracellular matrix, certain cell-cell adhesions, and the activation of many intracellular signaling pathways. Studies of LRs have primarily focused on mammals, while few studies of LRs in marine invertebrates have been reported. The functions of LRs in marine bivalve species are still unclear.

Methodology/principal findings

In this study, we cloned and sequenced an LR gene, MmeLR, from the clam Meretrix meretrix. The MmeLR mRNA and protein detected by realtime PCR and western blots were primarily distributed in muscle tissues. Far-western analysis showed a specific interaction between recombinant MmeLR and the LR ligand laminin. The results of the binding assay suggested a role of LR in cell adhesion and apoptosis in cultured primary cells of mantle tissues from M. meretrix. The Bcl-2 mRNA expression level in primary cells cultured in matrigel (mainly laminin) coated plates was significantly higher than in cells cultured in non-coated plates at 48 h of culture, while the p53 mRNA expression pattern was inversely related to that of bcl-2, suggesting that MmeLR is involved in p53-dependent apoptosis, and the binding between MmeLR and laminin inhibits apoptosis during primary cell culture.

Conclusions

Our results suggest that MmeLR may be involved in cell adhesion and apoptosis. This study may increase the understanding of the role of laminin receptor in cell adhesion and apoptosis and help to improve the culture of primary cells of marine invertebrates.

SUBMITTER: You Y

PROVIDER: S-EPMC3467220 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Publications

The potential roles of a laminin receptor in adhesion and apoptosis of cells of the marine bivalve Meretrix meretrix.

You Yanan Y Huan Pin P Wang Xiaomei X Liu Baozhong B

PloS one 20121009 10

<h4>Background</h4>The laminin receptors (LRs) play important roles in cell adhesion to the extracellular matrix, certain cell-cell adhesions, and the activation of many intracellular signaling pathways. Studies of LRs have primarily focused on mammals, while few studies of LRs in marine invertebrates have been reported. The functions of LRs in marine bivalve species are still unclear.<h4>Methodology/principal findings</h4>In this study, we cloned and sequenced an LR gene, MmeLR, from the clam M ...[more]

PMID: 23056594

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Project description:Hepatocellular carcinoma (HCC) is the most common primary cancer of the liver. Hence, new anti-liver cancer treatment strategies need to be urgently developed. Coptisine is a natural alkaloid extracted from rhizoma coptidis which exhibits anticancer activity in various preclinical models, including liver cancer. However, the molecular mechanisms underlying the anti-liver cancer effects of coptisine remains unclear. We used flow cytometry to assess the binding of coptisine to 67LR expressed on the surface of SMMC7721, HepG2, LO2 and H9 cells. Then SMMC7721, HepG2 and BEL7402 cells, belonging to the HCC cell lines, were treated with coptisine. The cell viability was detected using a cell counting kit-8 assay. Apoptosis was evaluated using flow cytometry and transferase-mediated dUTP nick-end labeling (TUNEL) assay. Apoptotic-related proteins and tumor death receptor 67-kDa laminin receptor (67LR) were detected using Western blot analysis. The cyclic guanosine 3',5'-monophosphate (cGMP) concentration was determined using enzyme-linked immunosorbent assay. sh67LR lentivirus, anti67LR antibody, and cGMP inhibitor NS2028 were used to determine how a 67LR/cGMP signaling pathway regulated coptisine-induced apoptosis. Tumor growth inhibited by coptisine was confirmed in a SMMC7721 cell xenograft mouse model. Coptisine selectively exhibited cell viability in human hepatoma cells but not in normal human hepatocyte cell line LO2 cells. Coptisine promoted SMMC7721 and HepG2 cell apoptosis by increasing 67LR activity. Both 67LR antibody and sh67LR treatment blocked coptisine-induced apoptosis and inhibition of cell viability. Coptisine upregulated the expression of cGMP. Moreover, cGMP inhibitor NS2028 significantly decreased coptisine-induced apoptosis and inhibition of cell viability. In vivo experiments confirmed that coptisine could significantly suppress the tumor growth and induce apoptosis in SMMC7721 xenografts through a 67LR/cGMP pathway. Coptisine-mediated 67LR activation may be a new therapeutic strategy for treating hepatic malignancy.

Project description:Simple Summary Numerous studies over the past few decades have revealed that the interactions of gastric cancer cells with laminins through integrins play important roles in tumor cell proliferation, infiltration, and metastasis. However, the association between gastric cancer cells and the laminin E8 fragment, which is the smallest integrin-binding component, has not been investigated. In this study, we revealed that the laminin 511-E8 fragment had a greater impact on the adhesion, morphology, and proliferation of gastric cancer cells than full-length laminin 511. Thus, the laminin 511-E8 fragment is considered to be suitable for investigating the interaction between gastric cancer cells and extracellular matrices in tumor invasion and metastasis. Further, the involvement of Cdc42 in the laminin 511-E8 fragment-induced enhanced adhesion of gastric cancer cells was suggested. Abstract Background: The interaction between cancer cells and laminin (Ln) is a key event in tumor invasion and metastasis. Previously, we determined the effect of full-length Ln511 on gastric cancer cells. However, the interactions between the Ln511-E8 fragment, a truncated protein of Ln511, and gastric cancer cells have not been investigated. Methods: We investigated the adhesion properties of gastric cancer cells to full-length Ln511 and Ln511-E8 fragments. Results: The proliferation of four gastric cancer cell lines (SH-10-TC, MKN74, SC-6-JCK, and MKN45) was highest on the Ln511-E8 fragment. Further, a larger cytoplasm was observed in SH-10-TC and MKN74 cells cultured on full-length Ln511 or Ln511-E8 fragments. The percentage of adhesive cells was highest on the Ln511-E8 fragment in all four cell lines. Moreover, adhesion of the gastric cancer cells to Ln511-E8 fragment-coated plates was reduced by the Cdc42 GTPase inhibitor in a dose-dependent manner, suggesting the involvement of Cdc42 in the Ln511-E8 fragment-induced enhanced adhesion of gastric cancer cells. Conclusions: The Ln511-E8 fragment had a greater impact on the adhesion, morphology, and proliferation of gastric cancer cells than full-length laminin. Thus, the Ln511-E8 fragment is suitable for investigating the interaction between gastric cancer cells and extracellular matrices in tumor invasion and metastasis.

Dataset Information

The potential roles of a laminin receptor in adhesion and apoptosis of cells of the marine bivalve Meretrix meretrix.

Background

Methodology/principal findings

Conclusions

Publications

The potential roles of a laminin receptor in adhesion and apoptosis of cells of the marine bivalve Meretrix meretrix.

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