Studies on the bone marrow microenvironment of hematological malignant patients

• Main Authors: Yung-Hsiu Lin, 林永修
• Other Authors: Wan-Fang Tzeng
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/73068828119513882857

碩士 === 輔仁大學 === 生命科學系碩士班 === 99 === Bone marrow is the main place for hematopoiesis. In bone marrow, stromal cells affect proliferation and differentiation of hematopoietic stem cells through secretion of cytokines or direct interaction. SDF-1α, secreted by stromal cells, interacts with CXCR4 on hematopoietic stem cells, and affects cell’s proliferation, mobilization and homing. SDF-1α promotes tumor growth, angiogenesis in tumor, and metastasis. SDF-1α has been widely studied in areas of hematopoiesis and tumor formation. Whether SDF-1α can affect the growth of bone marrow stromal cells is not understood. The stromal cells present low level of CXCR4 on the cell surface. Is it possible that SDF-1α enhances the growth of the stromal cells through autocrine signaling. NGAL (Neutrophil Gelatinase-Associated Lipocalin) affects cell's differentiation, proliferation and apoptosis. NGAL is secreted by skeletal muscle cells, epithelial cells and immune cells. The synthesis and secretion of NGAL in bone marrow was not clear. In this study, bone marrow mononuclear cells and stromal cells, bone marrow plasma and peripheral blood plasma from hematological malignant patients were used to understand the roles of NGAL in bone marrow. The results from MTT assay, β-galactosidase activity assay and the doubling time determination showed that there is no obvious difference in the third to the fifth generation of bone marrow stromal cells. After the fifth generation, some cells aging were faster, cells’ activity decreased obviously, β-galactosidase activity and cell doubling time increased. The addition of SDF-1α increased the growth of the senescent bone marrow stromal cells. During the time, the gene expression level of p53 decreased, and the levels of CXCR4 increased, the levels of bcl-2 and SDF-1α were not changed. The enhancement of SDF-1α was inhibited after addition of PI3K inhibitor, wortmannin. The result of ELISA showed SDF-1α in bone marrow plasma of patients were higher than that of normal. The secretion of SDF-1α in patient stromal cells was not always higher than that of normal cells. Although the level of NGAL in patients bone marrow plasma was higher than that of normal, the secretion of NGAL in patient stromal cells was not always higher than that of normal. The level of NGAL in bone marrow plasma was higher than that in peripheral blood plasma. The role of NGAL in bone marrow microenvironment needs further investigation.