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OBJECTIVE@#To investigate the expression of apoptosis related protein cellular Fas associated death domain like interleukin 1 converting enzyme inhibitory protein (c-FLIP) in peripheral blood mononuclear cells (PBMCs) of patients with rheumatoid arthritis and its relation with extrinsic apoptotic pathway.@*METHODS@#Sixty patients with rheumatoid arthritis were collected from Zhangzhou Affiliated Hospital of Fujian Medical University during January 2014 and June 2015, including 22 patients with low activities (DAS285.1). And 25 healthy controls were also collected. The mRNA and protein expression levels of c-FLIP and the extrinsic apoptotic pathway related proteins Fas-associated protein with death domain (FADD), caspase-8 in PBMCs were detected by real-time RT-PCR and Western blot, respectively. Correlations between c-FLIP and FADD, caspase-8 in PBMCs were analyzed by pearson test.@*RESULTS@#mRNA expression levels of c-FLIP, FADD and caspase-8 in PBMCs of patients with rheumatoid arthritis were all higher than those of healthy controls (all <0.05). mRNA expression levels of FADD and caspase-8 in patients with middle activities were significantly higher than those in patients with low activities (all <0.05), but the mRNA expression level of c-FLIP was not significantly higher than that in patients with low activities. mRNA expression level of c-FLIP in patients with high activities was higher than those in patients with middle or low activities (all <0.05), while the mRNA expression level of caspase-8 was lower than those in patients with middle or low activities (all <0.05). mRNA expression level of FADD in patients with high activities was higher than those in patients with low activities (<0.05). Pearson analysis showed that there was a positive correlation between c-FLIP and FADD mRNA expression (=0.323, <0.05), and negative correlation between c-FLIP and caspase-8 mRNA expression (-1.104, <0.05). The protein expression levels of c-FLIP and FADD in patients with middle activities were significantly higher than those in control group and patients with low or high activities (<0.05 or 0.01). The protein expression levels of caspase-8 in patients with middle and high activities were significantly higher than those in control group and patients with low activities (<0.05 or <0.01), and the protein expression level of caspase-8 in patients with high activities was higher than that in patients with middle activities (<0.05).@*CONCLUSIONS@#c-FLIP may be involved in the extrinsic apoptotic pathway in rheumatoid arthritis, and can provide reference for the evaluation of disease activities.
Subject(s)
Humans , Apoptosis , Genetics , Arthritis, Rheumatoid , Blood , CASP8 and FADD-Like Apoptosis Regulating Protein , Genetics , Gene Expression Profiling , Gene Expression Regulation , Leukocytes, Mononuclear , MetabolismABSTRACT
Objective To study the effects of COX-2 selective inhibitor celecoxib on the apoptosis of acute promyelocytic leuke-mia NB4 cell line,and to investigate its apoptosis mechanisms.Methods The expression of COX-2 mRNA in different cell lines was detected by reverse transcript polymerase chain reaction(RT-PCR).After treatment of NB4 with different doses of celecoxib,the in-hibition of NB4 growth was assayed by MTT,and the DNA fragmentation was examined by the DNA ladder test.The level of Bcl-2 protein expression was assayed by the flow cytometry.Results As compared with the no-medication treatment group,the DNA ladder fragments became more and more obvious after the treatment by different doses of celecoxib.The expression rates of Bcl-2 protein in the different doses of celecoxib groups (25,50,100 μmol/L)were (71.69 ±1.65 )%,(34.51 ±2.53)% and (29.28 ± 2.38)% respectively,compared with the Bcl-2 protein expression rate (85.34±2.89%)in the blank control group,the expression rate of Bcl-2 protein in different doses of celecoxib groups(50,100 μmol/L )was significantly decreased(P <0.05 ).Conclusion Celecoxib as COX-2 selective inhibitor could evidently induce the apoptosis of NB4 cells by down-regulating the expression of Bcl-2 protein in NB4 cells.
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Objective To study the clinical application of thymidine kinase 1(TK1) in diagnosis of common malignant tumors . Methods The serum TK1 levels of 444 cancer patients and 161 healthy individual(control group) were tested by western blot-en-hanced chemiluminescence assay .Results Compared with control group ,the levels of TK1 in patients with lung cancer ,gastrointes-tinal cancer or breast cancer were significantly higher(P<0 .05) .The positive rates of TK1 in lung cancer ,esophageal cancer ,colon cancer ,colorectal cancer ,breast cancer ,cervical cancer and ovarian cancer were respectively 52 .0% ,59 .6% ,62 .9% ,60 .0% , 62 .1% ,32 .3% ,21 .4% and 11 .0% .Compared with control group(the positive rate of TK1 was 0 .0% ) ,the positive rate of TK1 in lung cancer ,gastrointestinal cancer or breast cancer were significantly higher(P<0 .01) .Conclusion The test of serum TK1 is a valuable marker in common malignant tumor auxiliary diagnosis .
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Objective To investigate the relation between demethylation effect and apoptosis of valproate (VPA) in primary acute lymphoblastic leukemia (ALL) cells in vitro.Methods 10 cases of ALL patients was choosed to acquire leukemia cells.Cell growth curve were assessed by the MTT assay,the apoptosist of primary ALL was analyzed with DNA Ladder and Annexin-V-FITC/PI by flow cytometry.The expression methylation level of p15 was detected by hn-MSPCR,and p15mRNA were detected by RT-PCR.All dates was analyzed by SPSS16.0.Results The 50 ％ inhibition rate of VPA were 1.898 mmol/L to primary ALL cells assayed by MTT respectively.DNA ladder showed the apoptosis of primary ALL cells increased by adding VPA dose.Annexin-V-FITC/PI tests showed that the apoptosis percentage of primary ALL cells were (0.44±0.04) ％ in control group,(5.80±0.65) ％ in 1.0 mmol/L VPA group,(48.46±2.49) ％ in 2.0 mmol/L VPA group,(76.45±2.98) ％ in 4.0 mmol/L VPA group,the apoptosis percentage increased significantly (P ＜ 0.05).The demethylation of p15 INK4B gene decreased by adding VPA dose,the expression of p15 mRNA expression increased significantly compared with control group by RT-PCR (P ＜ 0.05).Conclusion It is found that VPA could induce demethylation of p15 INK4B gene,which could upregulate the p15 mRNA expression,due to the apoptosis of primary ALL cells.