亚硒酸化β乳你球蛋白诱导k562细胞凋亡及其机制研究-食品科学专业毕业论文.docx

AbstractThe Abstract The in vitro effects of seleno-p—lactoglobulin(se-p—lg)on the proliferation and apoptosis of human leukemia K562 cells were investigated．The changes of cell morphological，cell cycle，relevant cyclin protein and mRNA expression which were induced by se-p-lg were explored by MTT,HE staining，flow cytometry, immunofluorescence and real．time PCR etc． First of all，the proliferation inhibition and apoptosis of K562埘t11 Se—p—lg were researched．The MTT assays showed that se-13-19 significantly suppressed the growth of K562 cells in a dose··dependent and time-·dependent manner,while mouse C2C 1 2 myoblasts were viable after the same treatment．The cell growth inhibitory rate of K562 could reach 90％after exposure诵t11 1 00 I．tg／mL se-D-lg for 48h．HE and PI／Hoechst staining revealed that treatment of K562 cells、Ⅳitll se—p-lg appeared typical apoptotic characteristics such as cell volume reduction and chromatin shrinkage．The results of Annexin V—FITC／PI dyeing indicated that the apoptosis rate of K562 cells Was 1 8．75％ after cultured witll Se-p—lg for 48 h，then the rate increased to 40．76％when cultured for 72 h．FCM analysis demonstrated that treatment of K562 cells with Se-p-lg result in the accumulation of cells in the S and Gz／M phase． Apart from that，the signaling pathways of treatment of K562 cells晰th se-p-lg was studied．Immunofluorescence and elisa analysis uniformly revealed that the expression level of cyclin B 1 Was significantly down-regulated but the expression of p2 1 Was extremely up—regulated．Real-time PCR analysis showed that the mRNA expression of these proteins had the same trends and also showed that the mRNA expression of CDK4 Was decreased significantly． To sum up，se-p-lg could effectively inhibit the proliferation of k562 cells and induce apoptosis in vitro．It Was proved that se-p·lg would be a potential anticancer active material in future development． Key words：Seleno-p-lactoglobulin，K562 cells，leukemia cells，apoptosis，cyclinBl， P21 pr