OBJECTIVETo investigate the relationship of Fas mRNA and protein expression and apoptosis in human oral squamous cell carcinoma.

METHODSNorthern blot and flow cytometry (TUNEL method) were used to detect the expression of Fas mRNA and Fas protein, cell cycle and apoptotic level in oral squamous cell carcinoma. The relationship between Fas gene expression and OSCC apoptosis was analyzed statistically.

RESULTSFas mRNA and protein could be detected in all five normal oral mucosa specimens. There was positive correlation between expression of Fas mRNA/protein and cell differentiation as well as apoptosis in OSCC (P < 0.005).

CONCLUSIONThe expression of Fas gene was highly correlated with the differentiation and apoptosis in OSCC.

Apoptosis ; Carcinoma, Squamous Cell ; metabolism ; Humans ; Mouth Neoplasms ; metabolism ; fas Receptor ; metabolism

OBJECTIVETo examine the correlation between the mRNA and proteins expressions of gastrin(GAS), and the association of protein expression of GAS with apoptosis index(AI) and apoptosis regulation gene Fas/FasL, caspases in colorectal cancer.

METHODSThe expressions of GAS mRNA in tumor tissues of 79 cases with colorectal cancer were detected by nested RT-PCR. Cell apoptosis was detected by molecular biology in situ apoptosis detecting technic(TUNEL). Protein expressions of GAS, Fas/FasL, and caspases were detected by immunohistochemical staining (SP method).

RESULTSThe positive correlation was found between the mRNA and proteins expressions of GAS(rGAS=0.99, P<0.01). The mRNA and protein expressions of GAS in well and moderately differentiated cancers were significantly lower than those in poorly differentiated cancers (chi(2)(high vs low)=10.47, 10.23, P<0.01, chi(2)(middle vs low)=6.68, 4.95, P<0.05). The mRNA and protein expressions of GAS in papillary and tubular adenocarcinomas were significantly lower than those in mucinous adenocarcinomas, signet-ring cell carcinoma and undifferentiated carcinoma (chi(2)(papillary vs mucinous and signet-ring)=4.80, 6.22, chi(2)(papillary vs undifferentiation)=5.44, 8.43, chi(2)(tubular vs mucinous and signet-ring)=4.40, 4.38, chi(2)(tubular vs undifferentiation)=4.92, 6.43, P<0.05, respectively). The mRNA and protein expressions of GAS in Dukes' stages A, B were significantly lower than those in Dukes stages C, D (chi(2)=4.84, 4.45, P<0.01). The AI in GAS high and moderate expression groups of colorectal cancer were significantly lower than that in low expression group (q(high vs low)=6.71, q(middle vs low)=4.60, P<0.01). The positive expression rate of FasL was significantly different among GAS high, moderate and low expression groups of colorectal cancer (chi(2)=9.35, P<0.01). The positive expression rate of FasL in GAS high and moderate expression groups was higher than that in low expression group (chi(2)high vs low=6.24, chi(2)(middle vs low)=4.74, P<0.05).

CONCLUSIONSGAS plays an important role in the regulation of cell apoptosis in colorectal carcinoma, whose mechanism may be related to the aberrant expression of Fas/FasL. GAS will be one of the indicators of the biological behavior in colorectal carcinoma.

Adult ; Aged ; Caspases ; metabolism ; Colorectal Neoplasms ; metabolism ; pathology ; Fas Ligand Protein ; metabolism ; Female ; Gastrins ; metabolism ; Humans ; Male ; Middle Aged ; RNA, Messenger ; metabolism ; Young Adult ; fas Receptor ; metabolism

OBJECTIVETo explore the role of superoxide dismutase (SOD) coenzyme in regulation of Fas/FasL signal transduction and apoptosis of alveolar macrophages in pneumoconiosis patients.

METHODS50 male and Han nationality cases, including the dust exposed workers, Phase I, II pneumoconiosis patients confirmed by local pneumoconiosis diagnosis group according to GBZ 70 - 2002 diagnosis standard, who underwent whole lung lavage treatment were chosen as subjects. Their alveolar macrophages (AMs) were collected and purified. The cells were divided into three groups: the untreated group, the Fas/FasL group and the SOD group. 5 x 10(6) purified AMs were added into incubating bottles containing DMEM for 2 hours for purifying, added with SOD coenzyme and other block reagents separately, and then incubated for 24 hours in CO(2) incubation. The cells were harvested and lysed. Western-blot were used to analyze the expressions of Fas, FasL, Caspases-8 and Caspases-3. Software of Quantity One 7.0 was used to analyze the relative quantity of Fas, FasL, Caspase-8 and Caspase-3. TUNEL and DNA fragment analysis were used to analyze AMs apoptosis.

RESULTSThe apoptosis index in SOD coenzyme group (9.50 +/- 2.76)% and Fas/FasL group (14.01 +/- 2.56)% was significantly lower than that of in untreated group (19.18 +/- 2.83)% (P < 0.05). The catachrestic DNA ladder appeared in untreated group, was looming in Fas/FasL group, and was not found in the SOD group. The expressions of Fas, FasL, Caspase-8 and Caspase-3 of phase I and II in SOD group were higher than in the other two groups (P < 0.05). There was no significant difference in the expression of Fas, FasL, Caspase-8 and Caspase-3 among different phases of pneumoconiosis (P > 0.05).

CONCLUSIONSOD coenzyme can effectively regulate Fas/FasL signal transduction and block AMs apoptosis.

Adult ; Apoptosis ; Cells, Cultured ; Fas Ligand Protein ; metabolism ; Humans ; Macrophages, Alveolar ; metabolism ; pathology ; Male ; Middle Aged ; Pneumoconiosis ; metabolism ; pathology ; Signal Transduction ; Superoxide Dismutase ; metabolism ; fas Receptor ; metabolism

The aim of this study was to investigate the effects of the traditional Chinese medicine psoralen (PSO) plus long wave ultraviolet A (PUVA) on apoptosis in HL-60 leukemia cells and its mechanism. The effect of PUVA on HL-60 cell growth was assayed by MTT method and the changes of ultrastructure of cells were observed by electron microscopy. The apoptosis ratios, changes of mitochondrial membrane potential, expression of Fas and FasL protein and Fas and FasL mRNA were detected by FCM and fluorescent quantitation RT-PCR respectively. The expression of Caspase 8 and Caspase 3 protein were detected by immunocytochemistry (ICC). The results showed that the growth of HL-60 cells were inhibited by PUVA in time-and concentration-dependent manner through inducing cell apoptosis. When the irradiation time of long wave ultraviolet A lasted 15 minutes and the concentration of PSO was 80 microg/ml, the inhibition of HL-60 cell proliferation and apoptosis ratios reached the peak. There were obvious apoptotic ultrastructure changes and decrease of mitochondrial membrane potential in HL-60 cells after treatment with PUVA. The expression of Fas mRAN increased and expression of FasL mRNA decreased after treating with PUVA for 4 hours, and the same results of Fas, FasL expression on protein level were obtained also after treating with PUVA for 24 hours. The expression of caspase 8 and caspase 3 protein enhanced and reached the peak after treating with PUVA for 8 hours. It is concluded that the PUVA can inhibit the growth of HL-60 cells and induce apoptosis of these cells. The possible mechanism is supposed to be up-regulating Fas, down-regulating FasL levels and then activating the levels of caspase 8 and caspase 3. The decreasing of mitochondrial membrane potential may be involved in this process probably.
Apoptosis ; drug effects ; radiation effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Fas Ligand Protein ; metabolism ; Ficusin ; pharmacology ; HL-60 Cells ; Humans ; Ultraviolet Rays ; fas Receptor ; metabolism

OBJECTIVETo investigate the expression of Fas/FasL in infantile hemangiomas and discuss the role of Fas/FasL in the pathologic evolution of infantile hemangioma.

METHODThe EnVision immunohistochemical stain and RT-PCR technique was used to examine the expression of Fas/FasL protein and mRNA in the infantile hemangiomas.

RESULTS(1) In the early and middle proliferating stage, a number of infantile hemangioma cells expressed Fas. In the late proliferating stage, the number of positive cells increased obviously and the expression of Fas mRNA was reaching the strongest level. In the early regressing stage the Fas still existed in some cells and after that the expression decreased quickly. (2) Up to the middle proliferating stage, there were a few of FasL(+) cells foound. In the late proliferating stage, the number of FasL(+) cells increased significantly. From the early regressing stage, the number of FasL(+) cells decreased rapidly and disappeared.

CONCLUSIONThere may exist significant correlation between the expression of Fas/FasL and the development of the infantile hemangioma cells. The apoptosis of the infantile hemangioma cells mediated by Fas/ FasL may be the major reason of the spontaneous involution of infantile hemangioma.

Apoptosis ; Child ; Child, Preschool ; Fas Ligand Protein ; metabolism ; Hemangioma ; metabolism ; pathology ; Humans ; Hyperplasia ; Infant ; RNA, Messenger ; metabolism ; Signal Transduction ; fas Receptor ; metabolism

AIMTo investigate the effect of ligustrazine (LGT) on expression of Fas/FasL mRNA during pulmonary ischemia/reperfusion injury (PI/RI) in the rabbits.

METHODSSingle lung ischemia/reperfusion animal model was used in this study. The rabbits were randomly divided into three groups (n = 30, in each): sham operated group (Sham), I/R group (I/R) and I/R + LGT group (I/R + LGT). Changes of several parameters which included apoptotic index (AI), wet to dry ratio of lung tissue weight (W/D) and index of quantitative assessment of histologic lung injury (IQA) were measured at 1h, 3h, 5h after reperfusion in lung tissue. Meanwhile the location and expression of Fas/FasL mRNA were observed. Lung tissue was prepared for light microscopic and electron microscopic ob servation at 1 h, 3 h, 5 h after reperfusion.

RESULTSAs compared with group I/R, Fas/FasL mRNA slightly expressed in intima and extima of small pulmonary artery, alveoli, and bronchiole epithelia in group LGT. The values of AI, W/D and IQA showed significantly lower in group I/R + LGT than that in group I/R at 1 h, 3 h, 5 h after reperfusion in lung tissue (P < 0.01 and P < 0.05). Meanwhile, abnormal changes of the lung tissue in morphologically were lessen markedly in group I/R + LGT.

CONCLUSIONLigustrazine has notable protective effects on PI/RI in rabbits by inhibiting Fas/FasL mRNA express in lung tissue and decreasing apoptosis.

Animals ; Apoptosis ; Fas Ligand Protein ; metabolism ; Lung ; blood supply ; Lung Injury ; metabolism ; pathology ; Pyrazines ; pharmacology ; RNA, Messenger ; genetics ; Rabbits ; Reperfusion Injury ; metabolism ; pathology ; fas Receptor ; metabolism

The objective of this study was to investigate the apoptosis-inducing effect and underlying mechanism of naringenin (NGEN) on K562 cells in vitro. The inhibition of NGEN on K562 cells was evaluated by means of MTT assay so as to observe the cytotoxicity of NGEN; The morphological changes of the cells treated by NGEN were observed by transmission electron microscope; cell apoptosis rate influenced by NGEN was assessed by flow cytometry; the enzyme activity changes of caspase-3 and caspase-8 in the process of NGEN-induced K562 apoptosis were detected by Caspase Colorimetric Assay Kit; immunohistochemistry technique was used to detect FAS, FASL protein expression in K562 cells. The results showed that the growth of cells was inhibited by NGEN in dose-and time-dependent manners (p<0.05); NGEN-induced K562 cells apoptosis and sub-G1 peak was observed; some typically early and final phase changes of cell apoptosis were revealed under transmission electron microscope; the enzyme activity of caspase-3 and caspase-8 and the expression of FAS remarkably increased, meanwhile the expression of FASL was down-regulated (p<0.05). It is concluded that NGEN exerts anti-cancer effect by inducing K562 cell apoptosis, and the regulation of the expression of FAS and FASL. The caspase-3 and caspase-8 co-pathway brings about one of the mechanisms.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Fas Ligand Protein ; genetics ; metabolism ; Flavanones ; pharmacology ; Humans ; K562 Cells ; fas Receptor ; genetics ; metabolism

OBJECTIVETo detect the changes in the expression of apoptosis signals: Fas, FasL and NF-kappa B in the process of osteoclast-like cell (OLC) apoptosis effected by sodium fluoride.

METHODSAfter co-culture of osteoclast-like cells with 0, 5, 10 and 15 mg/L sodium fluoride, Fas, FasL and NF-kappa B antibody expressions were detected by immune-histochemistry.

RESULTSThe expression of Fas and FasL increased with the concentration of the sodium fluoride, however the expression of NF-kappa B decreased with the concentration of sodium fluoride.

CONCLUSIONIn the process of OLC apoptosis induced by sodium fluoride, the expression of Fas and FasL increased, and that of NF-kappa B decreased with the concentration of sodium fluoride respectively, and the changes of the expression present a dose-dependent pattern.

Animals ; Apoptosis ; Fas Ligand Protein ; Female ; Fluorides ; pharmacology ; Ligands ; Membrane Glycoproteins ; metabolism ; Mice ; Mice, Inbred C57BL ; NF-kappa B ; metabolism ; Osteoclasts ; cytology ; metabolism ; fas Receptor ; metabolism

CD40 Ligand ; pharmacology ; HL-60 Cells ; drug effects ; Humans ; fas Receptor ; metabolism

The purpose of this study was to investigate the expression of Fas, Fas ligand (FasL) and CD80 and function of FasL on the surface of acute myelomonocytic leukemia cells from WEHI-3 line. The expression of Fas, FasL and CD80 on the surface of WEHI-3 were detected by flow cytometry, the apoptosis of YAC-1 cell induced by FasL on the surface of WEHI-3 were detected by (3)H-TdR incorporation. The results showed that the expression rate of Fas, FasL and CD80 on the surface of WEHI-3 cells were (6.75 +/- 2.31)% (n = 5), (63.73 +/- 5.23)% (n = 5) and (5.06 +/- 0.41)% (n = 5) respectively. The apoptosis rate of YAC-1 cells (target cells) co-cultured with WEHI-3 cells (Effector cells) at the rate of 1:3, 1:10 and 1:30 were (26 +/- 4.5)%, (35 +/- 3.2)% and (43 +/- 2.7)% (n = 5) respectively. It is concluded that WEHI-3 cells have high expression of FasL and low expression of Fas and CD80 on their cell membrane, and can induce the apoptosis of Fas(+) YAC-1 cells.
Apoptosis ; physiology ; B7-1 Antigen ; biosynthesis ; Cell Membrane ; metabolism ; Fas Ligand Protein ; biosynthesis ; Humans ; Leukemia, Myelomonocytic, Acute ; metabolism ; pathology ; Tumor Cells, Cultured ; fas Receptor ; biosynthesis