Reversion Of The Expression Level Of LATS1Gene In BaLL-1Cell Line By Histone Methylation Inhibitor DZNeP

Objective: To research the expression levels and DNA promoter CpG islandmethylation、 histone methylator levels of LATS1gene in Acute LymphoblasticLeukemia patients and BALL-1cell line. Focuses on the effect of DZNeP that LATS1gene histone H3K9demethlation and mRNA expression increased,BALL-1cellapoptosis，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 clinicalpatients and certain malignant hematologic cell lines,（2） BSP were used to detect DNAmethylator level of LATS1gene in BALL-1cell line,（3）ChIP-qPCR were used todetect the effect of DZNeP reverse histon H3K9methylator and down regulating theexpression of LATS1gene,（4）Annexin V-FITC/PI to detect the effect that promotingBALL-1cell appotosis rate、cell cycle.Results:（1） Expression level of LATS1gene in AL patients is0.106times compare tohealthy people（RQ=0.115±0.074，p=0.000≤0.05），ALL is0.115times to normalperson（RQ=0.28±0.143，p=0.002≤0.05），LATS2is among the1.006times to thenormal people（RQ=1.006±0.041，p=0.990）. The lowest expression level ofLATS1/2in 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 has50CpG sites,the rate happened in BALL-1cell line is0.4%.LATS1gene histone H3K9methylaion in BALL-1(monomatyl and double methylp≤0.05）. Compare LATS1gene histone H3K9methylation level before to afterintervened by DZNeP in BALL-1cell line, we find the methylation isreversed(monomatylation P=0.000；dual mathylation P=0.003≤0.05）,and mRNAexpression of LATS1is up regulated. DZNeP can promote BALL-1cell apoptosis; withIC50concentration of DZNeP, BALL-1cell line cell cycle is arrested in G2/M period,accounted for6.98%（p≤0.05）.Conclusion: LATS1gene, which is one of the core members of the Hippo signalingpathway, show low expression level in acute leukemia patients.LATS2gene have no difference between AL patients to normal. This is closely related to the histone H3K9Monomethyl bismethyl. DZNeP can reversal LATS1histone H3k9methylation andincrease gene expression in BALL-1cell, contributing apoptosis, arrest the cell cycle atG2/M checkpoint. Finally, provides the experimental evidence for acute lymphoblasticleukemia gene therapy and epigenetic therapy.