沉默sirt2对急性髓系白血病耐药细胞增殖的影响内科学专业论文.docxVIP

中国医学科学院北京协和医学院硕士研究生学位论文Abstra 中国医学科学院北京协和医学院硕士研究生学位论文 Abstra ct Background：Acute myelocytic leukemia(AML)is kind of common malignant tumors of hematopoietic system results from uncontrolled proliferation，apoptosis and differentiation disorder of hematopoietic progenitor cell．Traditional chemotherapy method Can make the part of patients achieve remission，but it is difficult to obtain a significant therapeutic effect for relapse patients．It is particularly important to find a new therapeutic target to prolong patients’overall survival time．SIRT2 belongs to a member of the Sirtuins family，which is a member of the class III histone deacetylase family proteins．SIRT2 plays an important role in tumor cell proliferation，migration and invasion，etc．Studies have shown that SIRT2 expression significantly increased in AML patients compared with normal person，and it was higher in the relapse AML patients． SIRT2 may promote the development of AML，and induce the drugresistance．However， there limited data regarding the role of SIRT2 in AML． Objectives：We aimed to investigate the effect of SIRT2 gene on the proliferation of human multidrug resistance acute myeloid leukemia cell line HL60／A and its relevant molecular mechanisms to provide the basis for new therapeutic target ofAML． Methods：(1)The expression of SIRT2 in acute myeloid leukemia HL60 and multidrug resistance cells HL60／A were assayed by RT-PCR．SIRT2 shRNA plasmids were constructed，transfeeted into HL60／A cells and stable cell lines expressing shRNA were selected．(2)The expression of SIRT2 in the interference group and the control group were detected by RT-PCR and westem blotting．(3)Cell viability was detected by MTT assay．Colony formation ability was detected by Cell colony forming experiment．And cell cycle and apoptosis were detected by flow cytometry．(4)Western blotting technique was used to detect the cell cycle related proteins p2 1 and CyclinD 1，P13K／AKT signal pathway related proteins AKT and its phosphorylation le