Effect Of CHFR And PARP-1 On The Proliferation And Apoptosis Of B-cell Non-hodgkin's Lymphoma Raji Cells

Objective: To investigate the effect of checkpoint with forkhead-associated(FHA)and RING finger domains(CHFR)with poly(ADP-ribose)polymerase 1(PARP-1)on the proliferation and apoptosis of B-cell non-Hodgkin's lymphoma Raji cells.To overexpress the CHFR gene in Raji cells of non-Hodgkin's lymphoma by 5-Aza-dC and to inhibit the expression of CHFR gene in raji cells with lentiviral-mediated sh RNA.Then observe the changes of PARP-1,and Raji cells proliferation activity.Methods: 1.Raji cells were collected and randomly divided into blank group(no treatment).The experimental group(adding 5-Aza-dC reagent)was used to detect the expression of CHFR,PARP-1 mRNA expression level,Western blot detection of CHFR,PARP-1 protein expression.2.Raji cells were collected and randomly divided into three groups: blank group(no lentiviral vector),negative control group(negative control lentiviral vector),transfection group(lentiviral vector).Lentivirus was infected with Raji cells at different MOI.After 72 hours,the green fluorescence was observed by fluorescence microscopy to determine the best MOI = 15.3.The lentiviral vector was transfected into the negative group and transfection group with the optimal MOI.The three groups CHFR?PARP-1 expression were detected by q RT-PCR and western blot respectively.In accordance with CCK-8 kit instructions to detect blank group,negative control group,transfection group Raji cell proliferation.Flow cytometry was used to detect the apoptosis of Raji cells in blank group,negative control group and transfection group.Results:1.The mRNA and protein expression of CHFR in the 5-Aza-dC Raji group was significantly up-regulated compared with that in the control group.The CHFR mRNA levels were(1.00±0.0)?(1.613±0.101)(P<0.05)and protein were(0.0871±0.010)?(0.4984±0.008)(P<0.05).PARP-1 levels were decreased in mRNA levels and the protein expression levels compared with control group.The mRNA levels were(1.513±0.016)?(1.126±0.025)(P<0.05).The protein expression levels were(0.6825±0.012)?(0.3758±0.006)(P<0.05).2.After RAji cells were transfected with the Sh RNA vector for 72 h,The fluorescence demonstrated high transfection efficiency of over 80 %.Pre-experiment to verify the optimal MOI value = 15.The expression of CHFR gene in blank group,negative group and transfection group Raji cells was detected by q RT-PCR.The expression of CHFR gene was blank(1.00 ± 0.0),negative control group(0.936 ± 0.103),transfection group(0.607 ± 0.021),(P <0.05).The relative expression levels of PARP-1 mRNA were blank(1.237 ± 0.002),negative group(1.301 ± 0.010)and transfection group(1.485 ± 0.006)(P <0.05).Western blot analysis showed that CHFR expression was blank(0.8233 ± 0.003),negative control group(0.4111 ± 0.002),transfection group(0.1524 ± 0.003).transfection group was significantly lower than the blank group and the negative control group(P <0.05),The expression level of PARP-1 protein was blank group(0.3503 ± 0.002),negative control group(0.4468 ± 0.011),transfection group(0.8747 ± 0.001).The expression of PARP-1 in the transfection group was significantly higher than that in the blank group and the negative control group(P <0.05).3.After the silencing of CHFR gene,the proliferation ability of RAJI cells was significantly increased,and the apoptotic rate of RAJI cells was decreased: the control group(15.27±2.11)%;negative control group(20.23±1.8)%;sh RNA group(9.57±0.9)%.Conclusion:1?5-Aza-dC can overexpress CHFR gene and protein expression.2?lentivirus-mediated Sh RNA can down-regulate the expression of CHFR gene and protein expression in Raji cells.3?The 5-Aza-dC overexpression of CHFR gene,down-regulation of PARP-1 gene,cell proliferation and apoptosis rate increased significantly;silencing CHFR gene,PARP-1 gene expression High,cell proliferation rate increased,cell apoptosis was significantly decreased.4?CHFR might regulate the proliferation and apoptosis of B-cell non-Hodgkin's lymphoma Raji cells through PARP-1.5?CHFR gene,PARP-1 gene plays an important role in the proliferation and apoptosis of Raji cells,and can be used as a non-Hodgkin's lymphoma to treat potential targets.