The Expression And Mehtylation Of CHFR In B-cell Non-Hodgkin’s Lymphoma Cells

Objective:To investigate mRNA expression level of CHFR gene in B-cell Non-Hodgkin’s Lymphoma Cells, the effect of CHFR gene mathylation and methylation inhibitor on Raji cells proliferation and apoptosis, which can provide a theoretical and experimental foundation for Lymphoma clinical therapy.Methods:(pThe human Burkitt’s Raji lymphoma cells were cultivated in vitro. Raji cells were treated with1,4,7and10μmol/L of methylation inhibitor5-Aza-2’-deoxycytidine (5-Aza-2’-dC);②The CHFR gene expression level of lymph node of patients with non neoplastic diseases and Raji cells was detected by RT-PCR.③The CHFR gene expression level after treated with1,4,7and10μmol/L of5-Aza-2’-dC was detected by RT-PCR;④Methylation of CHFR gene of Raji cells and treated with10μmol/L of5-Aza-2’-dC was analyzed by MS-PCR;⑤The cells proliferation of Raji cells treated with different concentrations of5-Aza-2’-dC were analyzed by CCK assay.⑥he apoptosis of Raji cells was analyzed by flow cytometry.Results:①The CHFR mRNA relative expression of lymph node and Raji cells were0.898±0.07,0.121±0.013. CHFR gene had low-expression in Raji cells, the difference was statistically significant(P<0.05);②The relative expression of CHFR mRNA after treated by5-Aza-2’-dC at concentrations of1,4,7and10μmol/L were0.174±0.012,0.238±0.015,0.318±0.019,0.389±0.009, respectively, the difference was statistically significant(P<0.05).5-Aza-2’-dC can up-regulate the expression of CHFR mRNA by demethylation.③In untreated Raji cells CHFR gene was partly methylation and mathylation state was decreased after treated with5-Aza-2’-dC.④The growth inhibition rate of Raji cells after treated with5-Aza-2’-dC were(2.6±0.40)%,(8.8±0.96)%,(12.4±0.61)%,(16.2±1.1)%, respectively. The growth of Raji cells was inhibited. With the elevation of the drugs concentration, the inhibition rate increased. The difference was statistically significant(P<0.05).㏕he early apoptosis rate of Raji cells in the control group was (1.72±0.1)%. These cells which were treated with5-Aza-2’-dC were (3.86±0.35)%,(6.27±0.4)%,(9.57±0.33)%,(15.5±0.46)%. With the increasing concentrations of5-Aza-2’-dC, the apoptosis rate of Raji cells increased. There was statistically significant difference between these groups(P<0.05).Conclusion:①The expression of CHFR mRNA was down-regulated in Raji cells, which may contribute to the occurrence of B cell Non-Hodgkin’s lymaphoma.②The down-regulation of CHFR gene expression may be caused by methylation.③The expression of CHFR gene can be recovered by the methylation inhibitor. The up-regulation of CHFR gene expression can inhibit growth and promote apoptosis with concentration-dependent in Raji cells. The experiment may provide a theoretical and experimental foundation for Lymphoma clinical therapy.