Dataset Information

Metastasis-associated protein 1 induces VEGF-C and facilitates lymphangiogenesis in colorectal cancer.

ABSTRACT:

Aim

To study the correlation between high metastasis-associated protein 1 (MTA1) expression and lymphangiogenesis in colorectal cancer (CRC) and its role in production of vascular endothelial growth factor-C(VEGF-C).

Methods

Impact of high MTA1 and VEGF-C expression levels on disease progression and lymphovascular density (LVD, D2-40-immunolabeled) in 81 cases of human CRC was evaluated by immunohistochemistry. VEGF-C mRNA and protein expressions in human LoVo and HCT116 cell lines were detected by real-time polymerase chain reaction and Western blotting, respectively, with a stable expression vector or siRNA.

Results

The elevated MTA1 and VEGF-C expression levels were correlated with lymph node metastasis and Dukes stages (P < 0.05). Additionally, high MTA1 expression level was correlated with a large tumor size (P < 0.05). A significant correlation was found between MTA1 and VEGF-C protein expressions in tumor cells (r = 0.371, P < 0.05). Similar to the VEGF-C expression level, high MTA1 expression level was correlated with high LVD in CRC (P < 0.05). Furthermore, over-expression of MTA1 significantly enhanced the VEGF-C mRNA and protein expression levels, whereas siRNAs - knocked down MTA1 decreased the VEGF-C expression level.

Conclusion

MTA1, as a regulator of tumor-associated lymphangiogenesis, promotes lymphangiogenesis in CRC by mediating the VEGF-C expression.

SUBMITTER: Du B

PROVIDER: S-EPMC3063917 | biostudies-literature |

REPOSITORIES: biostudies-literature

ACCESS DATA

Similar Datasets

Project description:Background:Tumor necrosis factor alpha (TNF-?) enhances lymphangiogenesis in gallbladder carcinoma (GBC) via activation of nuclear factor (NF-?B)-dependent vascular endothelial growth factor-C (VEGF-C). Receptor-interacting protein 1 (RIP1) is a multifunctional protein in the TNF-? signaling pathway and is highly expressed in GBC. However, whether RIP1 participates in the signaling pathway of TNF-?-mediated VEGF-C expression that enhances lymphangiogenesis in GBC remains unclear. Methods:The RIP1 protein levels in the GBC-SD and NOZ cells upon stimulation with increasing concentrations of TNF-? as indicated was examined using Western blot. Lentiviral RIP1 shRNA and siI?B? were constructed and transduced respectively them into NOZ and GBC-SD cells, and then PcDNA3.1-RIP1 vectors was transduced into siRIP1 cell lines to reverse RIP1 expression. The protein expression of RIP1, inhibitor of NF-?B alpha (I?B?), p-I?B?, TAK1, NF-?B essential modulator were examined through immunoblotting or immunoprecipitation. Moreover, VEGF-C mRNA levels were measured by quantitative real-time polymerase chain reaction, VEGF-C protein levels were measured by immunoblotting and enzyme-linked immunosorbent assay, and VEGF-C promoter and NF-?B activities were quantified using a dual luciferase reporter assay. The association of NF-?B with the VEGF-C promoter was analysed by chromatin immunoprecipitation assay. A three-dimensional coculture method and orthotopic transplantation nude mice model were used to evaluate lymphatic tube-forming and metastasis ability in GBC cells. The expression of RIP1 protein, TNF-? protein and lymphatic vessels in human GBC tissues was examined by immunohistochemistry, and the dependence between RIP1 protein with TNF-? protein and lymphatic vessel density was analysed. Results:TNF-? dose- and time-dependently increased RIP1 protein expression in the GBC-SD and NOZ cells of GBC, and the strongest effect was observed with a concentration of 50 ng/ml. RIP1 is fundamental for TNF-?-mediated NF-?B activation in GBC cells and can regulate TNF-?-mediated VEGF-C expression at the protein and transcriptional levels through the NF-?B pathway. RIP1 can regulate TNF-?-mediated lymphatic tube formation and metastasis in GBC cells both in vitro and vivo. The average optical density of RIP1 was linearly related to that of TNF-? protein and the lymphatic vessel density in GBC tissues. Conclusion:We conclude that RIP1 regulates TNF-?-mediated lymphangiogenesis and lymph node metastasis in GBC by modulating the NF-?B-VEGF-C pathway.