家蝇幼虫抗菌肽对肿瘤细胞k562作用机制的初步分析-preliminary analysis of the mechanism of antibacterial peptides from housefly larvae on tumor cell k562.docxVIP

V VI 英文摘要 Preliminary experimental anticancer mechanism of Domestica larva antimicrobial peptides to tumor cells K562 Abstract Objectives: To isolate and purify the antimicrobial peptides from the Musca domestica larvae with inhibition activity to K562 cells in chronic myelocytic leukemia, and to observe whether damage the normal-human cells 293T (human renal epithelial cells). To observation the influence of the antimicrobial peptides purified to the K562 cell viability and the growth curve and to observation the influence of the antimicrobial peptides purified to the permeability of K562 cell membrane, in order to evaluate the optimal concentration of active play and time, and to evaluate the degree of the K562 cell membrane damage by the the antimicrobial peptides purified, and if the mechanism maybe directly killing; To observation the influence of the antimicrobial peptides purified to the nucleus and mitochondrial membrane potential and apoptotic proteins caspase (caspase-3) of K562 cells, and to explore the mechanism of antimicrobial peptides inhibiting K562 whether maybe to induce apoptosising. Methods Antimicrobial peptides of Musca domestica larvae with inhibition activity to K562 cells were sieved, through some processes that induced by acupuncture with E.coli induction, isolated and purified by trituration, centrifuga1ization, solid phase extraction (SPE) and reversed-phase high-performance liquid chromatography (RP-HPLC), to detect the activity to anti-tumor cells K562 by MTT colorimetric method and observed light microscope. To identify the protein nature of the peaks puried by comparing the RP-HPLC chromatogram when the wavelength are UV 214 nm and 280 nm, and to detect the effect to normal-human cells 293T by MTT colorimetric method. Recording the survival and the relative inhibition rate to make growth curve, just after treating with different concentrations of the antimicrobial peptides and different time to K562， stained by trypan b