Relationships Between β1, 6 Branching Of Cell Surfaces And Mouse Mammary Tumor Cells Behaviour Of Matastasis And Its Preliminary Research On Migration Mechanisms

In tumour cells, alterations in cellular glycosylation play a key role in their metastatic behaviour. Aberrant glycosylation can affect the cell regular function for example adhesion and migration and cell recognition.In the past decade advances in genomics, proteomics and mass spectrometry have enabled the association of specific glycan structures with disease states. The overexpression ofβ1,6 Branching on N-glycans are found in colon cancer and hepatoma tissues, which correlate with poor prognosis. This paper aims to explore the cell behavior differences between two mouse breast cell lines which has distinct level ofβ1,6 Branching and its underlying mechanisms.1.β1,6 Branching and cell behavior related with metastasis in 4TO7 cellThe RNA interference technology was employed to construct two cell lines 4TO7-siRNA3 and 4TO7-mock which have distinct levels of GnTV expression in the paper. We observed their cell behavior differences to identify whether the GnT-V and the N-glycansβ1,6 Branching of cell surface plays a causal role in 4TO7 cell adhesion, migration and invasion. First we designed three RNA interference plasmids and then transfected 4TO7 cell, finally picked up the monoclone. After testing the expression of GnT-V on RNA, protein and glycan level we chose the 4TO7-siRNA3 as the cell model. We have done the cell behavior experiments and found that the expression of GnT-V 4TO7-siRNA3 significantly decreased cell migration and invasion and enhanced the adhesion on FN compared with the negative control cells. Finally, the DNaseⅠand phalloidin staining indicated that the actin polymerization and depolymerization (actin dynamics) was inhibited in the 4TO7-siRNA3 cell model.2. Research on initial mechanisms of enhanced migration ofβ1,6 Branching high expression 4TO7 cellsThe initial mechanisms of variations of 4TO7-siRNA3 cell line was explored in the relationships between EGFR andβ1,6 branching. How the changes of glycans affect the EGFR and its downstream signaling was investigated. With the stimulation of EGF, the migration of 4TO7-mock cell has two stages. The rate of migration increased sharply after 8h. Then the expression of EGFR, the phosphorylation of EGFR and the several related signaling moleculars were investigated. The result is that the increasedβ1,6 Branching on the EGFR can enchance the phosphorylation of PKC-δand Erk and inhibit the activation of RhoA which is an important factor of cell migration in the second stage...