Effects of NF-?B decoy oligonucleotides on apoptosis in lung cancer cell A549 / 中国病理生理杂志

AIM: To investigate the effects of NF-?B decoy oligodeoxynucleotides (ODNs) on apoptosis in lung cancer cell A549. METHODS: The treatments of lung cancer cells (A549) were divided into three groups: group A (control group); group B (decoy ODN group) and group C (scramble decoy ODN group). FITC-labeled NF-?B decoy ODNs was transfected into A549 with LipofectAMINE TM 2000. The activation was observed by electrophoretic mobility shift assays (EMSA). The proliferation was observed by growth curve. The apoptosis of cells were observed by flow cytometry and TdT mediated dUTP-biotin Nick End Labeling (TUNEL). The expression of Bcl-2 and Fas were observed by Western blot. RESULTS: After FITC-labeled decoy ODNs was transfected for 1 hour, the decoy ODNs was detected in the nuclei of A549 cells. EMSA performed the depression of the NF-?B binding to the nucleus. The growth curve showed the inhibition of the A549 cell growth and the percentage of apoptosis was increased compare with control group by flow cytometry and TUNEL. The amount of apoptosis inhibitor (Bcl-2) in group A and group C were 2.0 times and 2.1 times more than that in group B, respectively. The level of apoptosis accelerator (Fas) in group B were 2.6 times and 2.3 times more than that in group A and group C, respectively via Western blot. CONCLUSION: The NF-?B decoy ODNs accelerate the apoptosis of lung cancer cell A549 and the mechanism may be due to its inhibiting the expression of Bcl-2 and increasing the level of Fas. [

Association of Fas promoter-670 polymorphism with systemic lupus erythematosus in southern Chinese / 中国病理生理杂志

AIM: To investigate whether Fas promoter-670 polymorphism is associated with systemic lupus erythematosus(SLE) in Southern Chinese. METHODS: 103 SLE patients and 110 controls were studied. Fas promoter -670 polymorphism was typed by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). RESULTS: No statistically significant differences were found when Fas promoter -670 genotype and allele frequencies were compared between the SLE and the controls. Similarly, no significant differences were seen between the male and female SLE and the controls, the SLE with lupus nephritis (LN) and the controls, the SLE with LN and the SLE without LN. CONCLUSION: Fas promoter -670 polymorphism does not appear to be associated with susceptibility to SLE in Southern Chinese. [

Quercetin regulates Fas expression and induces apoptosis in hepatocellular carcinoma HepG2 cells / 中国病理生理杂志

AIM:To investigate the role of 1,4,5-trisphosphate inositol(IP3)and Fas gene expression in apoptosis of HepG2 cells induced by quercetin.METHODS:HepG2 cells were treated with quercetin at different concentrations(including 20,40,60,80 ?mol/L)for 72 h and treated with 60 ?mol/L quercetin for 6 h,12 h,24 h,48 h and 72 h.IP3,Fas mRNA,Fas protein and apoptosis rate were assayed by IP3-3H Birtrak assay,RT-PCR,Western blotting and flow cytometry,respectively.RESULTS:When HepG2 cells were incubated with different concentrations of quercetin for 72 h,the IP3 content was lower than those in control.Fas mRNA expression,Fas protein expression and the apoptosis rate were higher than those in control.When HepG2 cells were incubated with quercetin for 6 h,12 h,24 h,48 h,72 h,the IP3 contents were lower than those in control incubated with 60 ?mol/L quercetin for 12 h.Fas mRNA expression was higher than that in control incubated with 60 ?mol/L quercetin for 12 h.Fas protein expression was higher than that in control.The apoptosis rate was significantly higher than that in control incubated with 60 ?mol/L quercetin for 24 h(P

Construction of chimeric gene expression vector inducing apoptosis of oral squamous carcinoma cells / 中国病理生理杂志

AIM: To gain cdcSTBX25A-fas chimeric gene bearing the regulative fragment cdcSTBX25A and opening read frame of fas in order to construct and identify eukaryotic expression vectors, pAdTrack-CMV-cdcSTBX25A-fas and pAdTrack-cdcSTBX25A-fas, which have the potential to transfer the tumor proliferative signal to promoting-apoptosis signal through up-regulate the fas expression by c-myc. METHODS: A pair of primers were designed according to the known sequences of cdcSTBX25A and fas. The cdcSTBX25A-fas chimeric gene was obtained by PCR reaction and inserted into the two plasmids pAdTrack-CMV and pAdTrack, respectively. The two recombinant plasmids were transferred into E. coli DH5?. The positive clones were screened in LB media with 50 mg/L kanamycin and identified by agarose gel electrophoresis, endonuclease digestion and PCR. The nucleotide sequence of inserted cdcSTBX25A-fas was determined by dideoxy chain termination method. Using software, BLAST was conducted to analyze the structure and sequence of the target fragments and compared with GenBank. RESULTS: The chimeric target gene, cdcSTBX25A-fas, of 1 603 bp was obtained. The positive host bacteria E. coli DH5? of recombinant plasmids were screened and amplified. The double-enzyme digestion showed the pAdTrack-CMV-cdcSTBX25A-fas and pAdTrack-cdcSTBX25A-fas presenting 9.2 kb, 1.6 kb bands, and 8.3 kb, 1.6 kb bands respectively. The sequence analysis confirmed that the two shuttle plasmids containing 1 597 bp cdcSTBX25A-fas with the ORF of fas. CONCLUSION: The eukaryotic expression plasmids pAd-Track-CMV-cdcSTBX25A-fas and pAdTrack-cdcSTBX25A-fas were successfully constructed.

Influences of dexamethasone on expression of fas, bcl-2 mRNA and apoptosis in eosinophils in vitro / 中国病理生理杂志

AIM: To investigate the effects of dexamethasone (DM) on expression of fas and bcl-2 mRNA and apoptosis in eosinophils (Eos) of bronchoalveolar lavage fluid (BALF) in asthmatic guinea pig. METHODS: The guineas pigs were stimulated with ovalbumin (OVA) for 48 hours. Hypodense Eos (HEos) and normodense Eos (NEos) were purified from BALF by gradients of Percoll. Eos was cultured in PRMI-1640 medium and DM (10 -10-10 -5 mol/L) was added. The mRNA of fas and bcl-2 in Eos was measured by hybridization. Apoptosis was detected by TdT-mediated dUTP nick end labeling (TUNEL). RESULTS: Eos was cultured under the condition of DM administration in vitro. Apoptotic Eos increased, fas mRNA expression increased and the bcl-2 mRNA expression decreased in a dose-dependent manner. There was significant correlation between Eos apoptosis and fas mRNA expression. The expression of bcl-2 mRNA was inversely correlated with Eos apoptosis. CONCLUSION: DM promoted Eos apoptosis, increased expression of fas and inhibited expression of bcl-2 in a dose-dependent manner in vitro. fas and bcl-2 might be one of mechanisms of DM promoting apoptosis in Eos.

Expression and significance of Fas,FasL,Bcl-2 and Bax in bladder cancer / 解放军医学杂志

Objective To investigate the relationship between the expression of Fas, FasL, Bcl-2 and Bax in transitional cell carcinoma of bladder and its clinical significance. Methods Immunohistochemistry method was used to detect the expression of Fas, FasL, Bcl-2 and Bax in 40 specimens of transitional cell carcinoma of bladder and 24 specimens of normal bladder mucosa tissue. Results The positive expression of Fas was detected both in transitional cell carcinonma of bladder and in normal bladder mucosa tissue, while FasL, Bcl-2 and Bax only in transitional cell carcinonma of bladder. The expression of Fas was significantly lower in careinoma of high-grade malignancy than in carcinoma of low-grade malignancy (P