The Effect Of Daintain/Allograft Inflammatory Factor-1 On Breast Cancer Cell Line MDA-MB-231 And MCF-7 Cells

Daintain was isolated and characterized from porcine intestine, and AIF-1(allograft inflammatory factor-1, AIF-1) was cloned from atherosclerotic allogenic heart grafts undergoing chronic transplant rejection, the protein of Daintain was identical with AIF-1, so named Daintain/AIF-1. it is an EF-hand-containing Ca2+ binding protein and scarfolld protein with 146 amino acid residues. Daintain/AIF-1 was reported to be distributed in multiple tissues, including liver, nervous system, spleen and lung. It was established that this protein was implicated in multiple pathophysiological processes such as immune response, regulation of blood glucose and cardiovascular diseases, autoimmune disease. In addition, this protein was associated with cell proliferation and cell migration.However, the characterization of Daintain/AIF-1 in breast tumor microenvironment has not been described. In order to explore the potential properties of Daintain/AIF-1, we mimic tumor microenvironment by incubating MDA-MB-231 and MCF-7 cells with macrophage-conditioned media or purified polypeptide Daintain/AIF-1, then assessed cell migration. On the other hand, we explored the directly effect of Daintain/AIF-1 on breast cancer cell, such as drug sensitivity and cell adhesion.The finished research work is listed below.(1) Construction of the stablly cell line which overexpress Daintain/AIF-1 in MDA-MB-231 and MCF-7 cells.(2) Construction of the stably macrophage cell line which siRNA knockdown Daintain/AIF-1 in U937 cells.(3) Conditioned medium of macrophages stimulated breast cancer cells and found that Daintain/AIF-1 can activate p38MAPK signaling pathway contributing to the upregulaion of TNF-a, consequently promote cell migration in breast cancer cells line MDA-MB-231 and MCF-7 cells.(4) Daintain/AIF-1 attenuate the drug sensitivity of breast cancer cells.(5) Daintain/AIF-1 upregulate 14-3-3ξprotein and reduce cell adhesion in MCF-7 cells.Our results indicated that Daintain/AIF-1 play a role in the progression of breast cancer.