دانلود رایگان مقاله تعاملات بین پروتئین ماکروفاژ اریتروبلاست و مهاجرت سلولی

ABSTRACT

Erythroblast macrophage protein is a novel protein known to mediate attachment of erythroid cells to macrophages to form erythroblastic islands in bone marrow during erythropoiesis. Emp-null macrophages are small with round morphologies, and lack cytoplasmic projections which imply immature structure. The role of Emp in macrophage development and function is not fully elucidated. Macrophages perform varied functions (e.g. homeostasis, erythropoiesis), and are implicated in numerous pathophysiological conditions such as cellular malignancy. The objective of the current study is to investigate the interaction of Emp with cytoskeletal- and cell migration-associated proteins involved in macrophage functions. A short hairpin RNA lentiviral system was use to down-regulate the expression of Emp in macrophage cells. A cell migration assay revealed that the relocation of macrophages was significantly inhibited when Emp expression was decreased. To further analyze changes in gene expression related to cell motility, PCR array was performed by down-regulating Emp expression. The results indicated that expression of mitogen-activated protein kinase 1 and thymoma viral proto-oncogene 1 were significantly higher when Emp was down-regulated. The results implicate Emp in abnormal cell motility, thus, warrants to assess its role in cancer where tumor cell motility is required for invasion and metastasis.

3. Results and discussion

Western blot analysis demonstrated that all three lentivirus-mediated shRNA transduction resulted in efficient down regulation of Emp. Results were very distinct in Emp#4 and Emp#6 shRNA lentivirus transduced cells (Fig. 1). Next, the effect of Emp down-regulation on cell motility was investigated. Quantification of cellular movements every 5 min up to 40 h showed that cell migration of Emp#4, Emp#6, and Emp#7 shRNA-lentivirus transduced cells were severely suppressed as compared to the control. Moreover, as the number of cells increased, results were more evident with time (Fig. 2). These results imply that cellular migration was significantly impaired following down regulation of Emp. Therefore, Emp can be considered as a novel molecule in cell migration. It can be a structural component of the actin cytoskeletal-adhesome complex or may be a crucial piece of signaling pathway machinery.