Experimental Study On Arsenic Trioxide Reversing Hypermethylation Of P16 And Activating Its Transcription In Malignant Lymphoma Cell Line CA46

Objective: To measure methylation level of gene p16 in the cell lines of six malignant hematonosis by DNA sequencing and Modified Methylation Specific PCR(MSP), i.e. Nested Methylation Specific PCR(nMSP), and then the screened hypermethylated cell line was used to study the mechanisms that how arsenic trioxide(As2O3) would reverse hypermethylation of p16 and exert its function of transcription regulation in the malignant lymphoma cell line --CA46. Methods: DNA sequencing and nMSP were used to screen the methylation level of cell lines. After the target cell line was identificated, SRB, growth curve as well as clone formation inhibition ratio methods were used to evaluate the effect that arsenic trioxide had on the ability of cell reduplication and cell vitality. Both methylation levels of CA46 before and after arsenic trioxide disposal were analyzed by nMSP; The expression of p16, DNA methyltransferase 1 (DNMT1), DNMT3A and DNMT3B were analyzed by RT-PCR; DNA ploid analytical method was used to analyze the effect that arsenic trioxide would bring on the cell life cycle of the target cell line. Results: 1. p16 gene in CA46 was found hypermethylated and is a choice option for pharmaceutical study on demethylation. The methylation level was apparently attenuated after 72h disposal of arsenic trioxide, and hypermethylation of p16 had been successfully reversed. 2. Expression of p16 gene in untreated group was mild while in the treated groups it had...