OBJECTIVETo study the apoptosis and mechanism of Fipronil on PC12 Cells.

METHODSThe effect of fipronil on the apoptosis and necrosis of PC12 cells of 3.13 x 10(-6), 1.25 x 10(-5) and 5.00 x 10(-5) mol/L three dose groups after 24 h treatment was detected by morphology and the apoptosis rate was detected by flow cytometer (FCM). The expression of Bcl-2 protein in PC12 cells of 3.13 x 10(-6), 1.25 x 10(-5) and 5.00 x 10(-5) mol/L three dose groups after 24 h treatment was measured by immunofluorescence.

RESULTSThe number of apoptotic cells of 3.13 x 10(-6) mol/L group was more than the control group examined by fluorescence microscope, and the number of dead cells of 5.00 x 10(-5) mol/L group was more than the control group. The apoptotic rates of PC12 cells was higher in the 3.13 x 10(-6) mol/L group than the control group measured by FCM, and the dead rates of PC12 cells was higher in the 5.00 x 10(-5) mol/L group than the control group (P < 0.05). Immunofluorescence cytochemistry experiment demonstrated that the level of Bcl-2 expression was significantly lower in the 3.13 x 10(-6) mol/L group than the control group.

CONCLUSIONAt low dosage, fipronil increases the apoptotic rates of PC12 cells possibly by decreasing the expression of Bcl-2 protein while at high dosage, fipronil only increases the amount of necrotic cells.

Animals ; Apoptosis ; drug effects ; PC12 Cells ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Pyrazoles ; toxicity ; Rats

OBJECTIVE: To analyze the prognostic value of BCL-2, BCL-6 and MYC in patients with diffuse large B cell lymphoma (DLBCL). METHODS: One hundred and sixty three cases of DLBCL in our hospital from March 2012 to March 2015 were selected. The specimens of lymphoma tissue of patients were collected. The expression of BCL-2, BCL-6 and MYC was detected by immunohistochemical method. The fusion of IGH/BCL-2, the gene breakage of BCL-6 and MYC were detected by interphase fluorescence in situ hybridization. The correlation of the expression levels of BCL-2, BCL-6 and MYC with the clinicopathological features and prognosis in the patients with DLBCL was further analyzed. RESULTS: MYC, BCL-2 and BCL-6 showed pale brown or reddish brown positive signals, among them MYC mainly positively expressed on the cell membrane, and BCL-2 mainly expressed on the cytoplasm and local cell membrane, and BCL-6 mainly expressed in the nucleus. The expression level of BCL-2 in ECOG physical status score 2 was higher than that in patients with ＜2 scores, and the expression level of BCL-2 in CD5 and germinal center B-cell-like (GCB) was significantly higher than that in patients with non-GCB (P＜0.05), and the international prognostic index (IPI) for 3-5 scores at the MYC expression level was significantly higher than that of the 0-2 score (P＜0.05); the expression level of BCL-6 in immune subtype CD5 and GCB was significantly lower than that in non-GCB (P＜0.05). The results of Cox multivariate analysis showed that the expression level of BCL-2, BCL-6 and MYC significant correlate with the overall survival and progression-free survival (P＜0.05) of the patients with DLBCL. CONCLUSION BCL-2, BCL-6 and MYC as important molecular markers are of high value for evaluating the prognosis of patients with DLBCL.
Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; metabolism ; Proto-Oncogene Proteins c-myc ; metabolism

Drastic surges in intracellular reactive oxygen species (ROS) induce cell apoptosis, while most chemotherapy drugs lead to the accumulation of ROS. Here, we constructed an organic compound, arsenical N-‍(4-(1,3,2-dithiarsinan-2-yl)phenyl)acrylamide (AAZ2), which could prompt the ROS to trigger mitochondrial-dependent apoptosis in gastric cancer (GC). Mechanistically, by targeting pyruvate dehydrogenase kinase 1 (PDK1), AAZ2 caused metabolism alteration and the imbalance of redox homeostasis, followed by the inhibition of phosphoinositide-3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway and leading to the activation of B-cell lymphoma 2 (Bcl2)/Bcl2-associated X (Bax)/caspase-9 (Cas9)/Cas3 cascades. Importantly, our in vivo data demonstrated that AAZ2 could inhibit the growth of GC xenograft. Overall, our data suggested that AAZ2 could contribute to metabolic abnormalities, leading to mitochondrial-dependent apoptosis by targeting PDK1 in GC.
Humans ; Signal Transduction ; Stomach Neoplasms/drug therapy* ; Reactive Oxygen Species/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Apoptosis ; Proto-Oncogene Proteins c-bcl-2 ; Cell Line, Tumor

OBJECTIVETo investigate the changes in SiHa cell apoptosis after inhibition of CD147 expression.

METHODSRNA interference (RNAi) technique was used to down-regulate CD147 expression in SiHa cells, and RT-PCR and Western blotting were used to detect expression of CD147, Bcl-2, Bim and caspase-3; the percentage of cell apoptosis were detected by flow cytometry.

RESULTSSiRNA sequence 1, 2 inhibited CD147 expression in SiHa cells effectively (P<0.05), resulting also in down-regulated expression of Bcl-2 (P<0.05) and up-regulated expression of caspase-3 and Bim(P<0.05). The percentage of apoptotic cells increased significantly, and early apoptosis was the most obvious in the cells (P<0.05).

CONCLUSIONSilencing of CD147 expression induces SiHa cell apoptosis partially through the Bcl-2 pathway .

Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Basigin ; metabolism ; Bcl-2-Like Protein 11 ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Down-Regulation ; Humans ; Membrane Proteins ; metabolism ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA Interference

Animals ; Bufo bufo ; Caspase 3 ; genetics ; metabolism ; Female ; Immunohistochemistry ; Male ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Spinal Cord ; metabolism ; Spinal Cord Injuries ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

OBJECTIVETo investigate the relationship between cell apoptosis and the quality of early mouse embryos, understand the significance of apoptosis-regulatory genes in early embryonic development, and explore a new approach to improving the embryo quality.

METHODSThe levels of cell apoptosis and proliferation in early mouse embryos in different developmental status (morphologically normal embryos, arrested embryos and fragmented embryos) were analyzed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), caspase in situ fluorescence and Bcl-2 immunofluorescence, and immunofluorescent detection of proliferating cell nuclear antigen (PCNA).

RESULTSThe cells in arrested embryos and embryonic fragments showed positive results in TUNEL assay with enhanced caspase activity and lowered expressions of Bcl-2 and PCNA.

CONCLUSIONCell apoptosis in early mouse embryos may be closely related to embryonic arrest and fragmentation.

Animals ; Apoptosis ; Caspases ; metabolism ; Embryo, Mammalian ; cytology ; Female ; Mice ; Mice, Inbred Strains ; Pregnancy ; Proliferating Cell Nuclear Antigen ; metabolism ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2

Microcystin is one of the monocyclic heptapeptides produced primarily by microcystis aeruginosa. Recent studies suggest that microcystin can induce cell apoptosis, as well as oxidative stress and mitochondrial alteration. Studies also indicate that Bcl-2 family and p53 may play an important role in the apoptosis induced by microcystin.
Animals ; Apoptosis ; physiology ; Humans ; Microcystins ; toxicity ; Microcystis ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

OBJECTIVE: To study the expression of apoptosis related genes, Bcl-2, bax, and iNOS in the olfactory epithelium of mice infected with influenza virus, and to discuss how they regulate apoptosis of the olfactory sensory neurons. METHOD: The expression levels of apoptosis related genes were detected with semi-quantity RT-PCR. RESULT: (1) The expression levels of Bcl-2 mRNA remain relatively constant after virus inoculation; (2) The expression levels of bax mRNA increased massively, and decreased gradually; (3) The expression levels of iNOS mRNA increased significantly in a short period, and then fell down to the undetectable level gradually. CONCLUSION The apoptosis related genes, Bcl-2, bax, and iNOS may play important roles in regulation of apoptosis of olfactory sensory neurons of mice infected with influenza virus.
Animals ; Mice ; Mice, Inbred BALB C ; Nitric Oxide Synthase Type II ; metabolism ; Olfactory Mucosa ; metabolism ; Orthomyxoviridae ; Orthomyxoviridae Infections ; metabolism ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; bcl-2-Associated X Protein ; metabolism

OBJECTIVEDuring the critical period of brain development, insufficiency of thyroid hormone results in severe mental retardation and learning deficit. This study was designed to investigate the effects of hypothyroidism on apoptosis and the expression of Bcl-2 and Bax gene in the developing rat hippocampus neurons and to explore the mechanism of brain development regulated by thyroid hormone.

METHODHypothyroidism was induced by administration of propylthiouracil (PTU, 50 mg/d) solution to the dams from gestational day 15 by gavage. Pups from both hypothyroid and control groups were harvested at postnatal day 1 (P1), P5, P10 and P15, respectively. Blood samples were collected at the time of death for the determination of thyroid hormone. Serum free tri-iodothyronine (FT(3)) and free thyroxine (FT(4)) were measured by using chemoluminescence. Hippocampus collected from the control and hypothyroid pups were examined under light and transmissional electron microscopy. Measurement of DNA fragmentation was carried out by agarose gel electrophoresis. The expression of Bcl-2 and Bax protein in the developing rat hippocampus neurons was performed by Western blotting.

RESULTSSignificantly lower circulating FT(4) and FT(3) levels confirmed the hypothyroid status of the experimental pups. The shrunken and contracted degenerations increased in hippocampus neurons of hypothyroid pups under light microscopy. Enhanced apoptotic cells were found in hippocampus neurons of hypothyroid pups under transmission electron microscopy, especially at P10 and P15. Extensive DNA fragmentation was seen throughout development in hippocampus of hypothyroid pups, but not in the euthyroid controls except for basal level at P10. The expression of Bcl-2 in the hippocampus neurons of hypothyroid pups was significantly lower than that of euthyroid controls at all stages of development (P1: 1.95 +/- 0.27 vs. 2.59 +/- 0.19, P < 0.05, P5: 1.86 +/- 0.24 vs. 2.47 +/- 0.17, P < 0.05, P10: 1.29 +/- 0.22 vs. 1.86 +/- 0.28, P < 0.05 and P15: 1.21 +/- 0.27 vs. 2.18 +/- 0.17, P < 0.01, respectively). The relative amount of expression varied significantly with age in the control pups. The level of Bcl-2 was high in hippocampus neurons of euthyroid at P1, P5, and decreased significantly at P10, and showed a trend of recovery at P15. Similar age-related variation in the expression of Bcl-2 gene was observed in the hypothyroid group at P1, P5 and P10, but the level was maintained low at P15. The expression of Bax in the hippocampus neurons of hypothyroid pups was significantly higher than that of control pups at all stages of development (P1: 1.69 +/- 0.14 vs. 1.24 +/- 0.23, P < 0.05, P5: 1.78 +/- 0.16 vs. 1.29 +/- 0.17, P < 0.05, P10: 1.92 +/- 0.18 vs. 1.45 +/- 0.14, P < 0.05 and P15: 1.86 +/- 0.14 vs. 1.51 +/- 0.12, P < 0.05, respectively). The ratio of Bcl-2/Bax in hippocampus neurons of hypothyroid pups was lower than that of age-matched controls (P1: 1.16 +/- 0.17 vs. 2.12 +/- 0.35, P < 0.05, P5: 1.05 +/- 0.16 vs. 1.94 +/- 0.36, P < 0.05, P10: 0.68 +/- 0.17 vs. 1.29 +/- 0.16, P < 0.05 and P15: 0.67 +/- 0.19 vs. 1.45 +/- 0.22, P < 0.01, respectively).

CONCLUSIONThyroid hormone significantly prevents apoptosis of hippocampus neurons. Congenital hypothyroidism increases not only the extent but also the duration of apoptosis by down-regulation of the anti-apoptotic gene Bcl-2 and maintaining a high level of the pro-apoptotic gene Bax.

Animals ; Animals, Newborn ; Apoptosis ; physiology ; Down-Regulation ; Hippocampus ; metabolism ; Hypothyroidism ; physiopathology ; Neurons ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; bcl-2-Associated X Protein ; metabolism

The aim of this study was to investigate the effect of TIEG1 on K562 cell apoptosis and expression of BCL-2/BAX, PTEN. The different concentration(0, 1, 5, 10, 20 ng/ml) of TIEG1 were used to treat K562 cells, the cell growth inhibition rate was detected by using MTT method. After treating K562 cells with 10.00 ng/ml TIEG1, the cell apoptosis was detected with flow cytometry. The RT-PCR was used to detected the expression levels of BCL-2 /BAX and PTEN. The results showed that TIEG1 displays inhibitory effect on proliferation of K562 cells in time-and dose-dependent manner (r = 0.52, P < 0.05) ; after K562 cells were treated for 6, 12, 24 and 48 h, the IC50 of TIEG1 were 48.19, 18.72, 9.5 and 3.85 ng/ml respectively. After treating K562 cells with 10.00 ng/ml TIEG1 for 0, 6, 12, 24, 48 h, the apoptosis rate were (2.13 ± 0.42)%, (7.79 ± 0.71)%, (11.17 ± 1.37)%, (24.66 ± 0.29)% and (48.60 ± 1.38)% respectively, and there was significant difference between groups(P < 0.05). In process of K562 cell apoptosis, the expression level of BCL-2 gradually decreased (r = 0.48, P < 0.05), meanwhile the expression levels of BAX (r = 0.69, P < 0.05) and PTEN (r = 0.57, P < 0.05) gradually increased. It is concluded that TIEG1 can indue apoptosis of K562 cells and inhibit K562 cell proliferation in time-and dose-dependent manner. In apoptosis process of K562 cells induced by TIEG1, the expression changes of BCL-2/BAX and PTEN associate with the K562 cell apoptosis.
Apoptosis ; Cell Proliferation ; Humans ; K562 Cells ; Kruppel-Like Transcription Factors ; metabolism ; PTEN Phosphohydrolase ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism