dznep逆转白血病ball1细胞株lats1基因表达的机制分析word格式论文.docx

基因启动子区组蛋白 H3K9 单甲基和双甲基化水平，进而上调该基因表达，并能将细胞周期阻滞于 G2/M 期，促进细胞凋亡。 关键词：急性白血病；表观遗传学 ；Hippo 信号通路；LATS 基因； DZNePReversion of the Expression level of LATS1 Gene in BALL- 1 cell line by Histone Methylation Inhibitor DZNePAbstractObjective: To research the expression levels and DNA promoter CpG island methylation、 histone methylator levels of LATS1 gene in Acute LymphoblasticLeukemia patients and BALL-1 cell line. Focuses on the effect of DZNeP that LATS1 gene histone H3K9 demethlation and mRNA expression increased,BALL-1 cellapoptosis，cell cycle changed. Provide a new target gene for the epigenetic treatment ofAcute Lymphoblastic Leukemia.Methods: （1）RT-PCR and WB were used to detect gene expression level in clinical patients and certain malignant hematologic cell lines,（2） BSP were used to detect DNA methylator level of LATS1 gene in BALL-1 cell line,（3）ChIP-qPCR were used todetect the effect of DZNeP reverse histon H3K9 methylator and down regulating the expression of LATS1 gene , （4）Annexin V-FITC/PI to detect the effect that promoting BALL-1 cell appotosis rate、cell cycle.Results:（1） Expression level of LATS1 gene in AL patients is 0.106 times compare to healthy people（ RQ= 0.115 ±0.074，p=0.000 ≤0.05），ALL is 0.115 times to normal person（RQ=0.28±0.143，p=0.002 ≤0.05），LATS2 is among the 1.006 times to the normal people（RQ=1.006 ± 0.04 1，p=0.990）. The lowest expression level of LATS1/2 in six hematologic malignancies cell lines is BALL-1（ RQ=0.28±0.143， p=0.000 ≤0.05）；（2）BSP text DNA promoter CpG island methylation rate ofLATS1 .The gene has 50 CpG sites ,the rate happened in BALL-1 cell line is 0.4%.LATS1 gene histone H3K9 methylaion in BALL-1(monomatyl and double methylp ≤0.05）. Compare LATS1 gene histone H3K9 methylation level before to afterintervened by DZNeP in BALL-1 cell line, we find the methylation is reversed(monomatylation P=0.000；dual mathylation P=0.003 ≤0.05 ）,and mRNAexpression of LATS1 is up regulated. DZNeP can promote BALL-1 cell apoptosis; with IC50