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Objective: To explore the possible mechanism of Fufangqizao Decoction in the treatment of gastric ulcer. Methods:Sixty rats were randomly divided into normal control group, ulcer control group, Fufangqizao decoction low-, medium- and high-dose groups (3 g/kg, 6 g/kg, and 12 g/kg); omeprazole group, with 10 in each group. Rat gastric ulcer model was prepared by acetic acid ablation method. The protein expressions of Foxo3a and Bim in gastric ulcer were detected by Western blot. Apoptosis of gastric mucosal epithelial cells was detected by TUNEL staining. Results: Compared with those in ulcer control group, the expression levels of Foxo3a and Bim in Fufangqizao decoction groups and omeprazole group were significantly lower, and the effect of high-dose Fufangqizao decoction on the expressions of Foxo3a and Bim was more significant than that of omeprazole (P<0.05). The apoptosis index in each group was (26.79±2.54)%, (22.41±3.67)%, and (14.38±3.40)%, which were significantly lower than that in ulcer model group (30.75±2.93)% (P<0.05). Conclusion: Fufangqizao decoction may inhibit gastric mucosal epithelial cell apoptosis by down-regulating Foxo3a/Bim expression and thus exerts its anti-gastric ulcer effect.
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Objective To determine the expression of miRNA-148a-3p in CD4+ T lymphocytes in peripheral blood of patients with psoriasis vulgaris,and to explore its role in occurrence of psoriasis vulgaris.Methods Totally,20 patients with psoriasis vulgaris and 20 healthy controls were enrolled from Guangzhou Institute of Dermatology between July 2017 and April 2018.Peripheral venous blood samples were obtained from these subjects,and CD4+ T lymphocytes were isolated from these peripheral blood samples by magnetic cell sorting system.Real-time quantitative PCR (RT-PCR) was performed to determine the expression of miRNA-148a-3p in CD4+ T lymphocytes in the peripheral blood.Potential target genes of miRNA-148a were predicted by using bioinformatics software,and verified by using a dual-luciferase reporter system.Western blot analysis was conducted to determine the protein expression of Bcl-2 interacting mediator of cell death (Bim,the potential target gene of miRNA-148a-3p) in the CD4+ T lymphocytes of the subjects.Statistical analysis was carried out with SPSS 20 software by two sample-t test for comparing the means of normally distributed data,and by Pearson correlation analysis for analyzing the correlation of two variables.If the data were not normally distributed,Mann Whitney U test was used for comparing means between two groups,and Spearman correlation analysis for analyzing the correlation of two variables.Results The miRNA-148a-3p expression in the CD4+ T lymphocytesin the psoriasis vulgaris group (18 cases,5.61 ± 1.66) was significantly higher than that in the healthy control group (12 cases,1.00 ± 0.26;U =12,P ＜ 0.05),and was positively correlated with the psoriasis area severity index (PASI) score (r =0.93,P ＜ 0.001).Bim was predicted to be one of the potential target genes of miRNA-148a-3p by bioinformatics software,which was also verified by using a dual-luciferase reporter system.The protein expression of Bim in the CD4 + T lymphocytes was significantly lower in the psoriasis vulgaris group (11 cases,0.69 ± 0.07) than in the healthy control group (8 cases,0.93 ± 0.06;t =4.38,P ＜ 0.01),and the protein expression of Bim in the patients with psoriasis vulgaris was negatively correlated with PASI score (r =-0.774,P ＜ 0.01).Conclusion miRNA-148a-3p is overexpressed in CD4+ T cells in the peripheral blood of patients with psoriasis vulgaris,which may regulate the protein expression of Bim,leading to abnormal activation of CD4+ T cells,and then participate in the occurrence and development of psoriasis.
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BACKGROUND: Hispolon has been shown to possess antitumor effects in various cancer cells. However, the underlying mechanisms are not fully understood. In this study, we evaluated the sensitizing effect of hispolon on TNF-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis in human renal carcinoma cells. METHODS: Apoptosis was analyzed by using cell-based cytometer. The mRNA levels were assessed by reverse transcription-PCR. Bax activation was determined by oligomerization and fluorescence-activated cell sorting with Bax-NT monoclonal antibody. The protein expression was measured by Western blotting. RESULTS: Hispolon induced up-regulation of Bim and death receptors expression at the post-translational level. CONCLUSIONS: Hispolon enhanced TRAIL-mediated apoptosis in renal carcinoma cells, but not in normal cells.
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Humans , Apoptosis , Blotting, Western , Flow Cytometry , Receptors, Death Domain , RNA, Messenger , TNF-Related Apoptosis-Inducing Ligand , Up-RegulationABSTRACT
Salvianolic acid A (SalA) is an effective compound extracted from traditional Chinese medicine Bunge. The Forkhead box O3a (FOXO3a) signaling pathway plays crucial roles in the modulation of ischemia-induced cell apoptosis. However, no information about the regulatory effect of SalA on FoxO3a is available. To explore the anti-cerebral ischemia effect and clarify the therapeutic mechanism of SalA, SH-SY5Y cells and Sprague-Dawley rats were applied, which were exposed to oxygen glucose deprivation/reoxygenation (OGD/R) and middle cerebral artery occlusion/reperfusion (MCAO/R) injuries, respectively. The involved pathway was identified using the specific inhibitor LY294002. Results showed that SalA concentration-dependently inhibited OGD/R injury triggered cell viability loss. SalA reduced cerebral infarction, lowered brain edema, improved neurological function, and inhibited neuron apoptosis in MCAO/R rats, which were attenuated by the treatment of phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) specific inhibitor LY294002. SalA time- and concentration-dependently upregulated the phosphorylation levels of protein kinase B (AKT) and its downstream protein FOXO3a. Moreover, the nuclear translocation of FOXO3a was inhibited by SalA both and , which was also reversed by LY294002. The above results indicated that SalA fought against ischemia/reperfusion damage at least partially the AKT/FOXO3a/BIM pathway.
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@#【Abstract】 【Objective】To investigate the mechanisms implicated in ataxia telangiectasia mutated(ATM)inhibitioncaused apoptosis in the cultured cerebellar granule neurons.【Methods】Primary cerebellar granule neurons(CGN)isolated from neonatal Sprague Dawley rats of 7-8 days were divided into the following groups:25 K group(survival group),5 K group(apoptosis group)and 25 K + KU-55933 treatment group(ATM inhibition group),25 K + KU-55933 + Mithramycin A treatment group(MMA group),25 K + KU-55933 + Chromonycin A3 treatment group(CMA3 group). The protein expression of p-ATM,ATM,Bim and Caspase 3 were detected by Western Blot. The apoptotic cells with nuclear pyknosis were detected by Hoechst-staining.【Results】Compared with 25 K group,the result of western blot showed that the protein expression of Bim and Caspase 3 were increased in the ATM inhibition group(P < 0.05). Compared with 25 K group,the nuclear pyknosis rate of 5 K group and ATM inhibition group were significantly increased(P < 0.05). Inhibition of Bim by Mithramycin A or Chromomycin A3 remarkably reversed ATM inhibition-caused apoptosis.【Conclusion】Inhibition of ATM induce Bim dependent apoptosis in cultured cerebellar granule neurons.
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RESUMEN La planificación de recursos humanos de un proyecto de construcción es una actividad de gran importancia para el desarrollo exitoso de este tipo de actividades. Resulta una labor zcompleja que requiere de herramientas y metodologías automatizadas que permiten la optimización de variables relacionadas con tiempo y costos. Building Information Modeling (BIM) es una base de datos digital que proporciona una réplica virtual del proceso constructivo a partir de cinco variables: i) el tiempo; ii) el costo; y tres dimensiones: x, y z; el modelo se conoce como BIM 5D. En este artículo se propone una metodología para la planificación de recursos humanos que tome como referencia la simulación del proceso constructivo BIM 5D. Se expone un conjunto de técnicas para la planificación del recurso humano en proyectos de construcción y se realiza la planificación de un caso de estudio a partir del enfoque BIM 5D. Con base en los resultados se formaliza un método para el diseño de la planificación del recurso humano. Comparado con otras metodologías, la propuesta presenta ventajas como la automatización del proceso y la posibilidad para la evaluación de distintas alternativas en tiempos reducidos.
ABSTRACT The human resources planning of a construction project is an activity of great importance for the successful development of construction activities. It is a complex task that requires automated tools and methodologies that allow the optimization of variables related to time and costs. Building Information Modeling (BIM) is a digital database that allows to obtain a virtual replication constructive process from five variables: i) time; ii) costs; and three dimensions x, y, z; model that is known as BIM 5D. In this paper it proposes a methodology for the planning of human resources based on the simulation of the BIM 5D construction processes. A set of techniques for the planning of the human resource in construction projects is presented and planning of a case study is carried out from the BIM 5D approach. Taking in account the results, it formalized a method for human resource planning. Compared to other methodologies, it presents advantages such as automation and the possibility of evaluating different alternatives in reduced times.
RESUMO O planejamento de recursos humanos de um projeto de construção é uma atividade de grande importância para o desenvolvimento bem-su-cedido deste tipo de atividades. É uma tarefa complexa que requer ferramentas e metodologias automatizadas que permitem a otimização de variáveis relacionadas com tempo e custos. Building Information Modeling (BIM), em português, Modelagem da Informação da Construção, é um banco de dados digital que fornece uma réplica virtual do processo de construção com base em cinco variáveis: i) tempo; ii) o custo; e três dimensões: x, y, z; o modelo é conhecido como BIM 5D. Este artigo propõe uma metodologia para o planejamento de recursos humanos que toma como referência a simulação do processo de construção do BIM 5D. Expõe-se um conjunto de técnicas para o planejamento de recursos humanos em projetos de construção e realiza-se o planejamento de um estudo de caso com base na abordagem BIM 5D. baseado nos resultados, formaliza-se um método para o planejamento de recursos humanos. Em comparação com outras metodologias, a proposta apresenta vantagens como automatização do processo e a possibilidade de avaliação de diferentes alternativas em tempos reduzidos.
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Objective The role of this essay is to explore the relationship of BIM deletion polymorphism and paclitaxel intrinsic resistance in breast cancer. Methods Human breast cancer cell lines were screened by techniques of PCR and gene sequencing to detect BIM deletion polymorphism.MTS was used to detect the cytotoxic effects of paclitaxel in different cell lines. Meanwhile,levels of BIM mRNA and protein were detected by rtPCR and Western Blot. Results Comparing with the wild type cell line(MCF7),T47D was a deletion cell line with BIM deletion polymorphism and resistant to paclitaxel(T47D vs.MCF-7,IC50>30le vs.IC50=0.16 vs.02 μmol/L). Moreover,the ratio of BIM EXON3 to EXON4 was significantly increased and the level of functional BIM protein was down-regulated in T47D compared with MCF7(P < 0.05). Conclusion BIM deletion polymorphism might initiate intrinsic resistance to paclitaxel therapy in breast cancer.
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Objective To observe the effects of serum containing Jinghou Zengzhi Recipe (JHZZ) on the expression of growth differentiation factor 9 (GDF9) and Bim in cumulus-oocyte complexes (COCs) and cumulus cells (CCs) of controlled ovarian hyperstimulation (COH) rats,and to investigate its curative effect and mechanism.Methods The rat COH ovarian model was prepared for collecting COCs and CCs,which were respectively co-cultured in vitro with the blank serum of female COH SD rat and serum containing JHZZ,and the each group was divided into 3 subgroups:the blank serum group (blank group),serum containing JHZZ group (control group) and serum containing JHZZ plus GDF9 receptor blocker group (experimental group),total 6 subgroups.COCs and CCs were collected after 24 h.The expression levels of GDF9 and Bim mRNA were detected by real-time quantitative PCR.The GDF9 protein expression levels were detected by Western blot.Results (1) The expression level of GDF9 mRNA in control group COCs was obviously higher than that in the control group and experiment group COCs (P＜0.05);the Bim mRNA expression level in COCs of control group and experiment group was significantly lower than that in COCs of blank group (P＜ 0.05);the GDF9 protein expression level in the control group COCs was significantly higher than that in the blank group COCs (P＜0.05),and also higher than that in the experiment group COCs,but the difference was not statistically significant (P＞0.05).(2)The comparison results of GDF9 mRNA expression level in CCs among 3 groups were consistent with those in COCs;the Bim mRNA expression level in CCs of control group and experimental group was significantly lower than that in the blank group (P＜ 0.05);the Bim mRNA expression level in the control group CCs was significantly lower than that in the experimental group CCs (P＜0.05);the GDF9 protein expression level in the control group CCs was significantly higher than that in the blank group and experimental group CCs (P＜0.05).(3)The GDF9 mRNA expression level in the control group COCs was significantly higher than that in CCs (P＜0.05),and the other inter-group comparisons between COSs and CCs had no statistical difference (P＞0.05).Conclusion Serum containing JHZZ can increase the GDF9 expression level in COCs and CCs,maintain the low expression of Bim in COCs and CCs,inhibit the ovarian cells apoptosis and promote the follicular development;COCs is more conducive to the expression of GDF9 mRNA compared with CCs eliminating oocyte.
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Objective To assess the effects of Jinghou Zengzhi Recipe(JHZZR),a Chinese prescription with the action of tonifying Qi and blood ,on the ovary apoptosis and expression of Bim in ovarian granulosa cells (GCs) of controlled ovarian hyperstimulation(COH) rats , and analyze the possible therapeutic mechanism. Methods A model of COH rats were prepaered and 30 female SD rats were randomly divided into 5 groups , including control group,positive control group,low,medium and high concentration group in six rats in each group. The apoptosis index(AI)in ovarian GCs were detected by TUNEL ,and the expression of Bim by qPCR. Results The AI of ovarian GCs in high and medium concentration group were obviously lower(P 0.05)comparing with control group. The mRNA levels of Bim were lower(P0.05)during the three concentration groups in Bim mRNA. Conclusion JHZZR can inhibit the ovarian GCs apoptosis of COH rats through decreasing the expression of Bim mRNA ,which improve the quality of ovarian follicle.
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Aim To investigate the effects of aspirin on Epstein-Barr virus (EBV)-transformed human B-lym-phocytes.Methods EBV-transformed human B-lym-phocytes were treated with certain concentrations of as-pirin.Cellular proliferation was analyzed by MTT as-say.Further evaluation of apoptosis of aspirin-treated cells was performed through light-field microscope, transmission electronic microscope(TEM),propidium iodide(PI)staining and flow cytometric analysis and DNA electrophoresis. Finally, immunoblot analysis was used to determine the expression levels of apopto-sis-associated proteins, proteins involved in mTOR pathway and PU.1 -Bim axis.Results Aspirin treat-ment inhibited proliferation of EBV-transformed human B-lymphocytes.We observed that aspirin treatment in-duced apoptosis in EBV-transformed human B-lympho-cytes,resulting in the decreased number and size of cells.Ultramicroscopic structural analysis via TEM in-dicated that aspirin treatment deformed the cellular nu-cleus,and led to peripheral chromatin and cytoplasmic vacuole.PI staining and flow cytometric analysis indi-cated that aspirin increased the permeability of cell membrane and decreased the viability of treated cells. Agarose electrophoresis revealed DNA smear in aspirin-treated cells.Mechanistically,mTOR signaling was in-hibited in aspirin-treated cells,as evidenced by the de-creased phosphorylation of S6K1 and S6 via immunob-lot analysis.Aspirin treatment led to the decrease of hematopoietic transcription factor PU.1 .Consequently, pro-apoptotic Bim, apoptosis-associated proteins caspase-3 and PARP were activated in aspirin-treated cells.Conclusion Aspirin may show anti-lymphoma effects via its inhibition of proliferation and induction of apoptosis of EBV-transformed human B-lymphocytes, in which mTOR signal pathway and PU.1 -Bim axis may be involved.
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Objective To study the expression level and function of micro RNA (microRNA)-218 in hepatocellular carcinoma (HCC) .Methods 46 cases of HCC surgery in the hepatobiliary surgery department of this hospital were selected and divided into the transfection group and nontransfection group .The expression ,proliferation and apoptosis of microRNA-218 and the expression level of B cell specific Maloney leukemia virus insertion site 1(Bmi-1) and cycling-dependented kinase 6(CDK6) in HepG2 cells were compared between the two groups .Results The expression level of microRNA-218 in HCC tissue was significantly lower than that in paracancerous tissues (P＜0 .05);the microRNA218 expression level was closely correlated with the clinicopathological characteristics such as tumor size and TNM stage(P＜0 .05);the HepG2 cell proliferation rates at 24 ,48 ,72 h after transfection in the transfection group were significantly lower than those in the nontransfection group(P＜0 .05);the HepG2 cell apoptosis rate in the transfection group was significantly higher than that in the nontransfection group(P＜0 .05);the Bim-1 and CDK6 expression levels after HepG2 cell transfection in the transfection group were significantly lower than those in nontransfection group(P＜0 .05) . Conclusion microRNA-218 can suppress the proliferation of HCC cells and promotes HCC cells apoptosis by down-regulating the Bim-1 and CDK6 expression level in potential targets .
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Lung cancer displays the highest morbidity and mortality worldwide. Non-small cell lung cancer (NSCLC) is the most com-mon type of lung cancer. In-depth research was performed on the pathogenesis and biological behavior of lung cancer and the im-provement of genetic testing level. The discovery of drugs targeting epidermal growth factor receptor and anaplastic lymphoma kinase plays a significant role in individual treatment of advanced NSCLC. BIM is a protein in the Bcl-2 family that promotes apoptosis, which leads to cell death. The BIM expression level and polymorphism can influence the therapeutic effect of targeted therapy and chemo-therapy on advanced NSCLC. Therefore, this review summarizes BIM and its effects on targeted therapy and chemotherapy for ad-vanced NSCLC.
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Objective To explore the effect of microRNAs 224 and 21 on human glioma stem cells survival and the possible molecular mechanisms.Methods qPCR was used to detect the dysregulated expression of microRNAs in malignant glioma samples, human GBM stem cells, artificially established GBM stem cell lines and human tissues.Caspase 3/7 assay, Annexin V apoptosis/fluorescence assay were performed to determine the effect of miR-21 or miR-224 mimics and inhibitor on cell apoptosis.Living cells count was used to assess miR-21 or miR-224 mimics and inhibitor on cell growth.TargetScan was used to explore potential targets of miR-21 and miR-224, and dual luciferase reporter assay was used to identify whether the 3’UTR of Caspase 3, Caspase 9 and Bim mRNA was a binding target of miR-21 or miR-224.Western blot was used to detect the expression of Caspase 3, Caspase 9 and Bim protein after transfection of miR-21 or miR-224 mimics or inhibitors.Results miR-21 and miR-224 are strongly upregulated in GSC samples, multiple GBM human tumor specimens, and GBM neurosphere stem cell lines ( P<0.05 ) .Caspase 3/7 assay and Annexin V apoptosis/fluorescence assay results showed that miR-224 and miR-21 regulated GSC apoptosis.Living cells count results demonstrated that miR-224 and miR-21 regulated GSC growth.miR-224 and miR-21 regulate pro-apoptotic gene expression by directly targeting Caspase 3, Caspase 9, and Bim 3’-UTRs. Conclusion These results indicate that miR-224 and miR-21 are important physiologic drivers of GSC resistance to apoptosis, providing new points of therapeutic leverage against these treatment-resistant cells.
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Objective To investigate the mechanism by which JAK2 inhibitor Ruxolitinib affecting the proliferation and apoptosis of human erythroleukemia leukemia(HEL) cells. Methods The HEL cells were treated with Ruxolitinib at different concentrations (1, 5, 10, 50, 100, and 500 nmol/L). Then the cell viability was detected by CCK-8 assay; the cell apoptosis was detected by Hochest staining method, the cell cycle was detected by flow cytometry, the mitochondrial membrane potential was assessed by rhodamine 123 with flow cytometry, and the Cysteine aspartic acid specific protease (Caspase)-3/7 protein activities were tested by kits. Moreover, the expression of JAK2 mRNA was measured by RT-PCR and the protein expressions of p-JAK2,p-ERK,Bcl-2 and Bim were observed by Western blotting analysis. Results After treated with different concentrations of Ruxolitinib at 1 nmol/L, 5 nmol/L,10 nmol/L, 50 nmol/L,100 nmol/L, and 500 nmol/L for 48 h, the HEL cell viabilities were (97.0±4.4)%,(92.0±3.9)%,(88.0±3.7)%,(81.0±3.1)%,(64.0±2.9)%,and (38.0±2.2)%, respectively. The ratio of high blue cells was significantly increased after treatment with 100 nmol/L Ruxolitinib for 48 h compared with the control group ([49.21±1.80]% vs [10.02±1.40]%, P<0.05). Flow cytometry showed G0/G1 phase cell ratio was higher after HEL cells exposed to 100nmol/L Ruxolitinib for 48 h compared with the control group ([73.1±3.6]% vs [45.2±3.0]%). The mitochondrial membrane potential was significantly decreased and Caspase-3/7 activity was significantly enhanced by treatment with Ruxolitinib at different concentrations (1-500 nmol/L) for 12 h and 24 h. RT-PCR showed that Ruxolitinib decreased JAK2 mRNA expression in a concentration-dependent manner in HEL cells after 48 h treatment.Western blotting analysis showed that the protein expressions of p-JAK2,p-ERK and Bcl-2 were significantly lower and Bim protein was significantly higher than those of control group(P<0.01). Conclusion Ruxolitinib may induce HEL cell apoptosis via JAK2 and ERK 1/2 pathways.
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Objective: To investigate the effect and interaction of microRNA-21 (miR-21) and Bcl-2 interacting mediator of cell death (BIM) on acquired drug resistance to epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) in human lung adenocarcinoma cells. Methods: The empty plasmid pcDNA3.1 and recombinant plasmid pcDNA3.1 -BIM were transiently transfected into gefitinib-resistant human lung adenocarcinoma PC9R cells, respectively, then the expression of miR-21 was determined by real-time fluorescent quantitative-PCR (RFQ-PCR), and the proliferation of PC9R cells treated with gefitinib was detected by cell counting kit-8 (CCK-8) assay. Meanwhile, the expression of miR-21 in PC9R cells was interfered by lentivirus infection method, then BIM expression and cell proliferation were detected by RFQ-PCR, Western blotting and CCK-8 assay, respectively. In addition, PC9R cells were transfected with recombinant plasmid pcDNA3.1-BIM and infected with lentivirus of miR-21 interference, then the sensitivity of PC9R cells to gefitinib was detected by CCK-8 assay. Results: After transfection with pcDNA3.1-BIM plasmids, the expression levels of BIM and miR-21 in PC9R cells were increased (P < 0.01). After infection of miR-21 expression interference lentivirus, the expression level of miR-21 and BIM in PC9R cells were down-regulated (both P < 0.05). Both knockdown of miR-21 and over-expression of BIM could improve the sensitivity of PC9R cells to gefitinib (both P < 0.05). Moreover, miR-21 knockdown combined with BIM over-expression could further improve the sensitivity of PC9R cells to gefitinib as compared with only miR-21 knockdown group (P < 0.05). Conclusion: MiR-21 and BIM may play key roles in the reversal of gefitinib-resistance to ECFR-TKIs, and they have a mutually antagonistic effect.
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Objective To investigate the molecular mechanism of the glucocorticoid?induced osteoblasts apoptosis,and to develop the effective intervention measures. Methods The MC3T3?E1 cells were treated with different concentrations of dexamethasone for 24 hours,then the apoptosis rate was detected with TUNEL analysis,the intracellular expression of Bim and Bax were determined by Westen blot. In addition,MC3T3?E1 cells were randomly divided into Bim?siRNA group,negative control siRNA group and control group. Twenty?four hours after transfection,10-4mmol/L dexamethasone was added to each group of cells for another 24 hours administration. The apoptosis rate was analyzed using the TUNEL,the mito?chondrial transmembrane electric potential was detected by flow cytometry after JC?1 staining,the expression of Bax,Bcl?2 in mitochondrial and Cyt?C,AIF in cytosolic were determined by Western blot. Results The rate of osteoblast apoptosis and Bim expression in cells were both significantly in?creased with the dosage of dexamethasone,there was no significant difference between the groups in the expression of Bax;the rate of osteoblast apop?tosis after the expression of silence Bim was significantly lower than the negative control siRNA group and control group,the expression of Bax and Cyt?C,AIF in cytosolic were significantly reduced,and the mitochondrial transmembrane electric potential was increased. Conclusion Bim is the key molecules in hormone?induced apoptosis of osteoblasts ,the expression of silence Bim can inhibit dexamethasone induced osteoblasts apoptosis through the mitochondrial pathway.
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To study the link between BRAFV600E status and the expression of BIM gene in papillary thyroid carcinoma( PTC) tissues and to analyze the association of these factors with clinicopathological characteristics. BRAFV600E status was determined by MASA-PCR, and qPCR was applied to detect the expression of BIM gene. Finally, the associations of these factors with clinicopathological characteristics were analysed. The rate of mutant BRAFV600E in PTC was 54. 1% , and the expression of BIM gene was lowered in BRAFV600E positive PTC tissues. Additionally, there was significant association( P < 0. 05) between BRAFV600E positiveness and raised TNM Staging (Ⅲ/ Ⅳ), and lowered BIM expression was significantly associated (P<0. 05) with the tumor size and raised TNM Staging(Ⅲ/ Ⅳ). These findings may help us to know more about the mechanism of PTC and to develop new diagnostic biomarkers or prognostic indicators of PTC.
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Aim To study the sensitivity of human gastric cancer BGC-823 cells to paclitaxel after trans-fection of SHIP2 ( The SH2 domain containing inositol 5-phosphatase 2 ) cDNA. Methods Apoptotic cells were determined by the propidium iodide method using flow cytometry. The levels of protein and mRNA ex-pression were measured by Western blot analysis and qRT-PCR, respectively. pCMV6-SHIP2 plasmid and empty vector were transiently transfected into BGC-823 cells, respectively. Stable cell lines were established after infecting BGC-823 cell with GV112-Puromycin and GV112-Puromycin-Bim( Bcl-2 interacting mediator of cell death) lentivirus particles. pCMV6-SHIP2 plas-mid was transiently transfected into the stable cell lines. Results BGC-823 cells were relatively insensi-tive to paclitaxel compared with SGC-7901 cells. The apoptotic rate was only (25. 6 ± 1. 6)% after the treat-ment with 0 . 3 μmol · L-1 paclitaxel for 48 h in BGC-823 cells. The expression levels of Bim protein and mRNA in BGC-823 cells treated with paclitaxel at dif-ferent time points were not significantly changed. The expression of Bim protein was increased after transfec-tion of pCMV6-SHIP2 plasmid, and the apoptotic rate was up to ( 50. 8 ± 0 . 9 )% in BGC-823 cells treated with paclitaxel for 48h. The expression of Bim protein was significantly inhibited after infecting with GV112-Puromycin-Bim lentivirus particles. The apoptotic rate of infected BGC-823 cells was only ( 27. 6 ± 1. 6 )%after treatment upon paclitaxel for 48h. Conclusion Overexpression of exogenous SHIP2 can increase the expression of Bim, induce apoptosis and enhance sen-sitivity of BGC-823 cells to paclitaxel.
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Objective To analyze the influence of intestinal trefoil factor(ITF) on Bim and Bcl-xl gene expression in neonatal rats with necrotizing enterocolitis(NEC),and to discuss the protective machanism of ITF on NEC.Methods Thirty neonatal rats were divided randomly into control group,NEC group and ITF group.NEC group were given intraperitoneal injection of saline 0.2 ml after NEC model of neonatal rats were established.ITF group were given intraperitoneal injection ITF 0.2mg after NEC model of neonatal rats were established.On the 4th day,all the subjects were put to death.We made HE stainting of the slice and made a histopathological examination and immunohistochemical method to detect Bim and Bc1-xl genes expression,and make image analysis.Results The pathological lesions indicated that intestinal tissue necrosis was severe in NEC group,which median was 3 point,but obviously lessen in ITF group,which median was 1 point,with ITF interfering.Image analysis showed the NEC group Bim gene expression (7.87 ± 0.14) higher than those in the control group (2.15±0.28) and ITF group (3.27±0.34),there were significant differences between 3 groups(P<0.05).Bcl-xl gene expression(11.23±0.22)in ITF group was higher than that in control group(1.89±0.28) and NEC group(2.51±0.13),there were significant differences between 3 groups(P<0.05).Conclusions Intestinal injury was ameliorated after ITF was injected intraperitoneally,ITF may protect the intestinal injury of neonatal rats with NEC by changing the Bim gene and Bc1-xl gene expresstion ratio.
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Aim: To investigate the expression and the regulation of Bim protein in leukemic cell of patient with chronic myelognous leukemia(CML). Methods: The primary leukemic cells from the peripheral blood or bone marrow of patients with CML were isolated and cultured. The expression of Bim in the present or absence of ima-tinib(BCR/ABL inhibitor) was detected by the Western blotting, immunohistochemical analysis and real-time quantitative PCR. Results: Bim was consecutively expressed in normal bone marrow, but significantly decreased in the bone marrow of patients with CML( P < 0. 05). Imatinib significantly enhanced Bim mRNA transcription and expression in the manner of time- and dose-dependence. Moreover, there was reverse correlation between Bim protein level and the percent of leukemic cell in bone marrow( r =0. 849 1, P <0. 05). Conclusion: Bim is the Bcl-2 family pro-apoptitic factor. The decreased expression of Bim could be vital in the CML, Bim protein up-expression following the suppression of BCR/ABL might be of importance in investigation of the Bim-induced ap-optosis for potential clinical therapy.