MicroRNAs （miRNAs） are small, noncoding RNAs which can often act as an oncogene or a tumor suppressor. Several miRNAs are associated with the development of hepatoceUular carcinoma （HCC）. We demonstrated that miR-...MicroRNAs （miRNAs） are small, noncoding RNAs which can often act as an oncogene or a tumor suppressor. Several miRNAs are associated with the development of hepatoceUular carcinoma （HCC）. We demonstrated that miR-125b significantly suppresses HCC cell proliferation and promotes apoptosis by inhibiting the gene expression of the anti-apoptotic protein, Bcl-2. Bioinformatic analysis indicated that the 3tUTR of Bcl-2 has binding sites for miR-I25b, Luciferase reporter assay confirmed the ability of miR-125b to dramatically suppress Bcl-2 transcription, suggesting that Bcl-2 is a target gene for miR-125b. We concluded that miR-125b acts as a tumor suppressor in hepatic tumor development by targeting Bcl-2 and inducing cancer celt apoptosis.展开更多
BACKGROUND:Many Chinese herbs,especially herbal injections,have been shown to have anti-tumor effects in recent years.However,since most reports focus on the clinical effectiveness of these herbs,their mechanisms of a...BACKGROUND:Many Chinese herbs,especially herbal injections,have been shown to have anti-tumor effects in recent years.However,since most reports focus on the clinical effectiveness of these herbs,their mechanisms of action are not well understood.In this study,we assessed apoptosis in the hepatocellular carcinoma(HCC)cell line HepG2 induced by an injectable extract from the seed of Coix lacryma-jobi(Semen coicis,SC),and monitored the expression of Bcl-2 and caspase-8.METHODS:Injectable SC was applied to HepG2 cells at different concentrations and the cells were collected 12,24 and 48 hours later.5-fluorouracil was used as a positive control group,and fluorescence-activated cell-sorting cytometry was used to measure the apoptosis rate of HepG2 cells and the expression of Bcl-2 and caspase-8 proteins.RESULTS:SC induced apoptosis in HepG2 cells in a concentration and time-dependent manner,and the expression of caspase-8 was elevated and prolonged.However,it did not significantly influence the expression of Bcl-2.CONCLUSION:Injectable SC may induce apoptosis in HCC cells by regulating the expression of caspase-8.展开更多
Objective:To investigate the relationship among Pokemon,NF-κB p65 and Bcl-2 in hepatoma cells.Methods:HCC cell HepG2,SMMC7721 and human fetal liver cell line L02 cells were used,and expression of Pokemon,NF-κB p65 a...Objective:To investigate the relationship among Pokemon,NF-κB p65 and Bcl-2 in hepatoma cells.Methods:HCC cell HepG2,SMMC7721 and human fetal liver cell line L02 cells were used,and expression of Pokemon,NF-κB p65 and Bcl-2 in three cells were detected by realtime PCR and western blot.Then siRNA of Pokemon was applied to inhibit the expression of Pokemon and NF-κB p65 and apoptotic rate was determined by flow cytometric analysis. Results:Expressions of Pokemon,NF-κB p65 and Bcl-2 in human hepatoma cell HepG2, SMMC7721 expression were significantly higher than those in human embryonic stem cells L02. siRNA of Pokemon inhibited the expression of Pokemon,NF-κB p65 and Bcl-2 in liver cancer cells,and significantly increased apoptosis of liver cells.While siRNA of NF-κB n65 inhibited the expression of NF-κB p65 and Bcl-2,but Pokemon expression in hepatoma cells had no significant change.Conclusions:The proto-oncogene Pokemon can inhibit PI4ARF by specific transcription regulation of cell cycle and can induce tumors.In addition,Pokemon can regulate NF-κB p65 through the expression of apoptosis repressor,and promote the development of liver cancer.It suggests signal network in the liver include the regulation of new non-classical NF-κB regulatory pathway.展开更多
Objective: The expression of B-cell lymphoma 2(Bcl-2) seems to be influenced by the endocrine environment. Numerous reports demonstrate the diverse expression of Bcl-2 family members under sex steroid regulation. With...Objective: The expression of B-cell lymphoma 2(Bcl-2) seems to be influenced by the endocrine environment. Numerous reports demonstrate the diverse expression of Bcl-2 family members under sex steroid regulation. With the exception of estrogen-related tumors, androgen-related tumors have shown their characteristics in Bcl-2 expression. In this study, the status of Bcl-2 expression in male hepatocellular carcinoma(HCC) patients was examined to verify the high incidence of HCC in males.Methods: Tumor tissue microarray was used to examine Bcl-2 expression levels in 374 HCC cases including 306 males and 68 females. Kaplan-Meier method, log-rank test, and Cox proportional hazards model were applied to investigate the predictive value of Bcl-2 in HCC patients.Results: Immunohistochemistry analysis showed that male patients with higher Bcl-2 levels had significantly longer median survival time and recurrence time than those with lower levels. However, no significant differences in outcomes were found between different Bcl-2 levels in female patients. When the male patients were stratified into several age points, the level of Bcl-2expression showed poorer predictive efficiency in the 45–49 and 55–60 age groups in andropause-age patients compared with other age groups. Bcl-2 was an independent prognostic factor for both overall survival(P < 0.0001) and recurrence time(P =0.0001) in male patients. After excluding male patients in the 45–60 age group, the predictive efficiency was enhanced(n = 147,OS, P = 0.0002, TTR, P < 0.0001).Conclusions: Bcl-2 expression is an independent predictor of survival and recurrence in male HCC. Bcl-2 levels may also be regulated by androgens or androgen receptors in male HCC patients. Bcl-2 levels change and exhibit poor predictive efficiency when androgen levels vary dramatically(andropause age).展开更多
Objective: To correlate the frequency of p53 mutations, bcl-2 expression and the proliferation status (proliferating cell nuclear antigen, PCNA) in patients with bladder cancer with cell proliferation, ...Objective: To correlate the frequency of p53 mutations, bcl-2 expression and the proliferation status (proliferating cell nuclear antigen, PCNA) in patients with bladder cancer with cell proliferation, apoptosis and their clinico-pathologic ?ndings. Methods: Para?n-embedded sections from 39 super?cial (T1G1-G3) and 23 invasive (T2-T4a G3 N0M0) primary transitional cell carcinomas (TCC) in the bladder were investigated immunohistochemically for p53, bcl-2 and PCNA. The median follow-up was 37 months; 24 had recurrences. The proliferation index (PI) was expressed as a percentage of the PCNA-positive cells in the tumor cells. Apoptosis was detected by terminal deoxy-nucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and the apoptotic index (AI) was expressed as a percentage of the TUNEL- positive tumor cells. Results: P53 mutation was identi?ed in 50 patients (80.6%). The mutation was most common in tumors of grade 3 (91.3%) as compared to grade 2 (78.5%) and grade 1 (72.7%, P<0.05). Stage pT2 tumors had a higher frequency of p53 mutation (95.7%) as compared to pTa-1 tumors (74.3%, P<0.01). Only 14 tumors (22.5%) expressed bcl-2; grade 3 tumors expressed bcl-2 signi?cantly more frequently (P<0.05); there was no correlation between bcl-2 and tumor stage. There was no interrelation between p53 mutation and bcl-2 expression (P>0.05). The PI ranged from 17.2% to 41.8% (median 22.4%) and the AI from 1.9% to 3.5% (median 2.9%) in bladder cancer. Statistical analyses revealed a close associations between PI, AI and tumor grade and stage of bladder cancer. Conclusion: P53 mutation correlates with invasion. P53 and PCNA overexpression may o?er valuable additional prognostic information in bladder tumors. With the progression of the tumor grade, cell proliferation may be accompanied by frequent apoptosis in bladder cancer, but the PI increased much more than the AI.展开更多
Objective:To explore the influence of silencing Bcl-2 expression by small interfering RNA（siRNA） on Bcl-2 protein expression,cell apoptosis rale and radiosensilivity of gastric cancer BCC823 cells.Methods:siRNA segm...Objective:To explore the influence of silencing Bcl-2 expression by small interfering RNA（siRNA） on Bcl-2 protein expression,cell apoptosis rale and radiosensilivity of gastric cancer BCC823 cells.Methods:siRNA segment for Bcl-2 gene was designed and synthesized,then was induced into gastric cancer BGC 823 cells by liposome transfection.Bcl-2 protein expression was detected by Western Blotting.After X radiation,flow cytometry and clone forming assay were used to determine the effects of RNA interference on BGC823 cell apoptosis rate and radiosensitivity. Result:After the transfection of Bcl-2 siRNA,the positive expression rate of Bcl-2 protein in BGC823 cells was（35.45±2.35）%.Compared with the control group and negative siRNA transfection group,the rate was significantly decreased（P【0.01）.The apoptosis rate of BGC823-RNAi cell was（10.81±0.91）%,which was significantly higher than the control group and negative siRNA transfection group（P【0.01）.After 48h X radiation,the apoptosis rate of BGC823-RNAi was（28.91±1.40）%,which was significantly higher than the control group and the group without radiation （P【0.01）.During clone forming assay D<sub>0</sub>,D<sub>4</sub> and SF<sub>2</sub> values in Bcl-2 siRNA1 transfection group were all lower than those in the control group.The radiosensitivity ratio was 1.28（the ratio of D<sub>0</sub>） and 1.60（the ratio of D<sub>4</sub>）.Conclusions:Specific siRNA of Bcl-2 gene can effectively inhibit the expression of Bcl-2 gene,enhance the radiosensitivity and apoptosis of gastric cancer BGC823 cells,having good clinical application perspective.展开更多
Mitochondrial K＋-ATP （mito-KATP） channels play an important role in cellular function and survival following ischemic stress. The present results revealed that intervention with diazoxide, a mito-KATP channel opene...Mitochondrial K＋-ATP （mito-KATP） channels play an important role in cellular function and survival following ischemic stress. The present results revealed that intervention with diazoxide, a mito-KATP channel opener, led to an increase in Bcl-2 expression in the cerebral cortex of rats subjected to cerebral ischemia reperfusion injury. In addition, the intervention also led to clear improvements in neuronal mitochondrial morphology and consciousness post-injury. Glibenclamide a mito-KATP channel blocker, exhibited the converse effects. Both diazoxide and glibenclamide exerted dose-dependent effects （in particular, at 18 mg/kg diazoxide and 25 mg/kg glibenclamide）. These findings suggest that diazoxide exerts a neuroprotective effect on cerebral ischemia reperfusion injury by opening mito-KATP channels and upregulating Bcl-2 expression.展开更多
AIM: To examine the expression of thymidylate synthase (TS) and oncoprotein Bcl-2 in advanced colorectal cancer (CRC) patients,and to determine their mutual relationship,association to therapeutic response and impact ...AIM: To examine the expression of thymidylate synthase (TS) and oncoprotein Bcl-2 in advanced colorectal cancer (CRC) patients,and to determine their mutual relationship,association to therapeutic response and impact on disease outcome.METHODS: Tumor samples from 67 patients with CRC,who were treated at advanced stage with either irinotecan alone or in combination with 5-fluorouracil/ leucovorin,were analyzed for expression of TS and Bcl-2 using immunohistochemistry.RESULTS: A significant linear correlation between lower expression levels of Bcl-2 and lower levels of TS expression was found (P = 0.033).Patients with high levels of both TS and Bcl-2 expression had a significantly longer disease-free survival (DFS) (42.6 mo vs 5.4 mo,n = 25) than those with low TS/Bcl-2 index (P = 0.001).Tumors with low levels of both TS and Bcl-2 were associated with a longer survival with metastasis (WMS) interval in the whole patients group (n = 67,P = 0.035).TS/Bcl-2 index was not significantly related to disease-specific survival.CONCLUSION: The present data suggest that CRC patients with low TS/Bcl-2 demonstrate a significantly shorter DFS and longer WMS.展开更多
We tested the effects of curcumin treatment on a rat model of Alzheimer＇s disease induced by beta-amlyoid （Aβ1-40） expression. We investigated alterations in the expression of the apoptosis-related genes Bax and B...We tested the effects of curcumin treatment on a rat model of Alzheimer＇s disease induced by beta-amlyoid （Aβ1-40） expression. We investigated alterations in the expression of the apoptosis-related genes Bax and Bcl-2 in the hippocampus, as well as changes in the spatial memory and cognitive function of the rats. Reverse transcription-polymerase chain reaction and immunohistochemistry results showed that Bax expression was remarkably decreased and Bcl-2 expression was increased in the rat Alzheimer＇s disease model after curcumin treatment. Morris water maze results showed that the average time of escape latency was shortened in the curcumin treated model rats. Our study shows that curcumin can significantly improve spatial learning and memory functions in rats with Aβ1-40-induced Alzheimer＇s disease by modulating Bax and Bcl-2 expression.展开更多
Brain-derived neurotrophic factor was utilized in the present study to treat cell injury models induced by aggregated β-amyloid（25 35）. Methylthiazolyldiphenyl-tetrazolium bromide assay and western blot analysis sh...Brain-derived neurotrophic factor was utilized in the present study to treat cell injury models induced by aggregated β-amyloid（25 35）. Methylthiazolyldiphenyl-tetrazolium bromide assay and western blot analysis showed that brain-derived neurotrophic factor provided neuroprotection against cellular apoptosis by suppressing the decline in β-amyloid（25 35）-induced cell activity and the increasing ratio of Bax/Bcl-2. After treating pheochromocytoma cells with tyrosine kinase receptor B receptor inhibitor K252a, brain-derived neurotrophic factor reverses the above- mentioned changes. The experimental findings suggested that brain-derived neurotrophic factor prevented β-amyloid peptide-induced cellular apoptosis by modulating Bax/Bcl-2 expression, and this effect was associated with binding to the specific tyrosine kinase receptor B receptor.展开更多
The ventral tegmental area and the locus coeruleus are associated with psychological and physical dependence of opioid addiction. To date, very little is known about brain-derived neurotrophic factor （BDNF） and Bcl-...The ventral tegmental area and the locus coeruleus are associated with psychological and physical dependence of opioid addiction. To date, very little is known about brain-derived neurotrophic factor （BDNF） and Bcl-2 gene and protein changes following morphine addiction. The present study utilized immunohistochemistry and in situ hybridization techniques, which revealed that there were increased BDNF levels, but decreased Bcl-2 levels in the prefrontal cortex, locus coeruleus, hippocampus, and the ventral tegmental area during morphine-dependence formation and abstinence. However, the levels of BDNF remained unchanged, and Bcl-2 expression was increased in the nucleus accumbens. These results showed that BDNF and Bcl-2 are involved in the development of morphine dependence, and precipitation of abstinence syndrome.展开更多
BACKGROUND： Estrogen has been clinically demonstrated to attenuate ischemic brain injury. However, the precise mechanisms remain controversial. OBJECTIVE： To investigate the effects of estradiol on angiopoietin-1 mR...BACKGROUND： Estrogen has been clinically demonstrated to attenuate ischemic brain injury. However, the precise mechanisms remain controversial. OBJECTIVE： To investigate the effects of estradiol on angiopoietin-1 mRNA and Bcl-2 expression, as well as apoptosis and cerebral blood flow, in ovadectomized rats with focal cerebral ischemia following reperfusion. DESIGN, TIME AND SETTING： Randomized, controlled, animal experiment. The study was performed at the Central Laboratory, Chongqing Medical University from September to December 2005. MATERIALS： Estradiol benzoate was purchased from Shanghai Ninth Pharmaceutical Factory; corn oil was purchased from Walmart Supercenter; TUNEL kit, rabbit anti-rat Bcl-2 polyclonal antibody, and biotin-labeled goat anti-rabbit antibody were purchased from Wuhan Boster, China. METHODS： Healthy, female, 6-month-old Wistar rats-wild-type and estrogen alpha receptor gene knockout （ERKO）-were randomly divided into estradiol and control groups with 25 animals in each group. The rats were intramuscularly injected with estradiol benzoate （100 μg/kg per day） at 30 days following bilateral ovariectomy or corn oil （1 mL/kg per day） for seven consecutive days. Following administration, cerebral ischemia/reperfusion models were established using the right middle cerebral artery occlusion （MCAO） method. After 30 minutes of MCAO, estradiol and control groups were separately injected with estradiol benzoate and corn oil with the above-mentioned doses. MAIN OUTCOME MEASURES： Cell apoptosis was determined by TUNEL; angiopoietin-1 mRNA and Bcl-2 gene expression was determined, respectively, by immunohistochemical staining and RT-PCR. In addition, changes in cerebral blood flow were measured by laser Doppler flowmetry. RESULTS： Changes in angiopoietin-1 mRNA and cerebral blood flow in estradiol-treated, wild-type, MCAO rats following ischemia/reperfusion were greater than in control rats （P 〈 0.01 or 0.05）. However, no significant difference was observed between estradiol-treated ERKO MCAO rats and control rats. In addition, estradiol-treated wild-type and ERKO MCAO rats exhibited significantly increased Bcl-2 expression （P 〈 0.05） and decreased number of apoptotic cells in brain tissues compared with control groups （P 〈 0.05）. CONCLUSION： Estradiol upregulated angiopoietin-1 mRNA and Bcl-2 expression, suggesting that estradiol might be involved in protective mechanisms of cerebral ischemia/reperfusion injury.展开更多
Macrophage cells play an important role in the initiation and regulation of the immune response. All-trans retinoic acid (ATRA) and its natural and synthetic analogs (retinoids) affect a large number of biological pro...Macrophage cells play an important role in the initiation and regulation of the immune response. All-trans retinoic acid (ATRA) and its natural and synthetic analogs (retinoids) affect a large number of biological processes.Recently , retinoids have been shown promise in the therapy and prevention of various cancers. However, many interesting questions related to the activities of retinoids remain to be answered: (Ⅰ) Molecular mechanisms by which retinoids exert their effects; (Ⅱ) why the clinical uses of retinoids give undesirable side effects of varying severity with a higher freqllency of blood system symptoms; (Ⅲ)little is known for its impacts on macrophage cells etc. We set up this experiment, therefore, to examine the apoptosis of ATRA on macrophage Ana-1 cell line. Apoptosis of the cells was quantitated, after staining cells with propidium iodide (PI), by both accounting nuclear condensation and flow cytometry. When the cells were treated with ATRA at or higher than 1 μM for more than 24 h, significant amount of the apoptotic cells was observed. Induction of apoptosis of Ana-1 cells by ATRA was in time- and dose-dependent manners, exhibiting the similar pattern as the apoptosis induced by actinomycin D (ACTD). ATRA treatment of Ana-1 cells also caused the changes of the mRNA levels of apoptosis-associated gene bcl-2, as detected by Northern blot analysis. The temporal changes of bcl-2 expression by ATRA was also parallel to that by ACTD. In conclusion,ATRA can induce apoptosis in macrophage cells, which may be helpful in understanding of immunological functions retinoids.展开更多
Objective: To investigate the relationship between oxidative stress and apoptosis and bcl-2 expression following traumatic brain injury (TBI). Methods: Male Sprague-Dawley rats were subjected to lateral fluid percussi...Objective: To investigate the relationship between oxidative stress and apoptosis and bcl-2 expression following traumatic brain injury (TBI). Methods: Male Sprague-Dawley rats were subjected to lateral fluid percussion brain injury (FPBI) of moderate severity. U-74389G (20 mg/kg) were administered intravenously before FPBI. The neurological functions were measured by beam-walk task (BWT) and beam-balance task (BBT). In addition to morphological evidence of apoptosis, TUNEL histochemistry was used to identify DNA fragmentation in situ with both light and electron microscopic levels. The internucleosomal fragments of DNA in apoptotic cells were examined using agarose gel electrophoresis. Bcl-2 protein expression was detected by immunohistochemistry. Results: The scores of BWT and BBT were significantly improved (P<0.01) in the treated animals. The treatment significantly reduced the number of apoptotic cells that was counted in the areas of the injured hemisphere at various time points following TBI. No DNA ladder was detected in the treated rats. Bcl-2 expression was observed in the cerebral cortex, subcortical white matter, dentate gyrus, hippocampal CA1 and CA3 region ipsilateral to injured hemisphere. Bcl-2 positive cells displayed normal nuclear morphology; Little Bcl-2 positive cells revealed morphological feature of apoptosis or necrosis. The immunoreactivity of Bcl-2 protein decreased significantly in the hippocampus ipsilateral impact site as early as 6 h post-injury. During 1-3 d after injury, the bcl-2 protein expression decreased relatively slow. In the U-74389G treated groups, the downregulation of bcl-2 expression was halted. Conclusion: In this model, apoptosis is associated with an activation of lipid peroxidation. U-74389G may block oxidative stress and halt the downregulation of bcl-2 expression. These may be one of the molecular mechanisms of the neuro-protective effects by U-74389G.展开更多
Objective: To study the effect of auricular acupuncture on dysmnesia and the relationship between the memory improvement and bcl 2 protein expression in vascular dementia (VD) rats. Methods: Forty Wistar rats were ran...Objective: To study the effect of auricular acupuncture on dysmnesia and the relationship between the memory improvement and bcl 2 protein expression in vascular dementia (VD) rats. Methods: Forty Wistar rats were randomized into control group, VD group, acupuncture+VD group and pseudo operation group, with 10 cases in each group. Rat VD model was established by using 4 vessel occlusion method. Otopoint "Nao" point and "Shen"(MA SC) were punctured, once daily continuously for 15 days. The rats’ memory capability was tested with Y maze method and bcl 2 expression of the brain tissues displayed by immunohistochemical method and measured using MIAS 2000 Image Analyzer. Results: Results showed that the scores of control group, VD group and acupuncture+VD group before operation were 5.68±1.29, 6.07±1.67 and 5.86±1.74 respectively, while following auricular acupuncture treatment, the scores of the 3 groups were 5.81±1.51, 18.06±2.68 and 8.31±1.85 separately, suggesting that the VD rat’s learning and memory abilities in acupuncture+VD group were raised apparently in comparison with those of VD group (P<0.01). In control, VD and acupuncture+VD group, bcl 2 immuno reaction positive neurons in CA 1 area of the hippocampus were 14.31±4.87, 28.67±5.63 and 65.74±8.19 respectively, displaying that the improvement of learning and memory abilities caused by auricular acupuncture treatment may be related to the up regulation of bcl 2 expression (an inhibitory gene of apoptosis). In comparison with control group, the loss of neurons in the pyramidal cell layer of the hippocampal CA 1 area of VD group was more severe, while that of acupuncture group was markedly lighter. Conclusion: Auricular acupuncture of otopoint "Nao" point and "Shen"(MA SC) can raise the learning and memory abilities of VD rats, which may be realized by its inhibitory effect on apoptosis and the protection action on ischemic hippocampal neurons.展开更多
Objective: To study the expression of Fas and Bcl-2 proteins on T lymphocyte subsets in the peripheral blood of relapsing patients with condyloma acuminatum(CA) and healthy controls.Methods: Flow cytometry (permeabiza...Objective: To study the expression of Fas and Bcl-2 proteins on T lymphocyte subsets in the peripheral blood of relapsing patients with condyloma acuminatum(CA) and healthy controls.Methods: Flow cytometry (permeabization and staining procedure with conjugated antibodies) was used.Results: We observed that the expression of Fas protein on CD4^+ T lymphocyte subset of CA patients was significantly higher than that of healthy controls( P<0.01 ).Conclusions: Increased expression of Fas proteinon CD4^+ T lymphocyte subset may be a cause of de-creased percentage of CD4^+ T lymphocyte subset. This induces the increased ratio of CD4^+/CD8^+.展开更多
Objective: To evaluate the anti-tumor effects of SeO2 and its mechanisms on three human lung cancer cell lines. Methods: Three lung cancer cells A549, GLC-82 and PG were treated with 3-30 μmol/L SeO2. Flow cytometry ...Objective: To evaluate the anti-tumor effects of SeO2 and its mechanisms on three human lung cancer cell lines. Methods: Three lung cancer cells A549, GLC-82 and PG were treated with 3-30 μmol/L SeO2. Flow cytometry was used to detect apoptosis, and analyze the changes of expression of p53 and Bcl-2, as well as ROS and Ca2+ level within cells. Results:SeO2 markedly inhibited cell proliferation and viability, and prompted apoptosis after 48 h treatment. SeO2 at 10 μmol/L induced 47.8% apoptosis in A549 cells, 40.8% in GLC-82 cells, 18.2% in PG cells. SeO2 at 30 μmol/L induced 37.8% apoposis in PG cells,but did not increase apoptotic raes in other two cells. SeO2 could down-regulate the mean fluorescent intensity of Bcl-2 from 65.8 to 9.6 in A549, but not in GLC-82 and in PG cells, up-regulate wild type p53 level in all three cells. SeO2 decreased the ROS and Ca2+ level markedly within three tested cells. Conclusion: SeO2 showed anti-tumor effect via apoptosis pathway in three lung cancer cell lines. The decrease of ROS and Ca2+ level within cells as well as regulation of Bcl-2 and p53 expression may play important roles in above apoptotic procedure.展开更多
To evaluate the apoptosis positivity, the expression of Bcl-2, bax proteinsin 30 patients with squamous cell cervix carcinoma before and after radiotherapy. Methods: By usingimmuno-histochemical and TDT-dUTP nick end ...To evaluate the apoptosis positivity, the expression of Bcl-2, bax proteinsin 30 patients with squamous cell cervix carcinoma before and after radiotherapy. Methods: By usingimmuno-histochemical and TDT-dUTP nick end labelling techniques, 30 cases of squamous cell cervicalcarcinoma were analyzed. Results: The apoptosis positivity before and after irradiation was 76.7%and 100% respectively, with the difference being significant (P 【 0.05); The positive rates of Bcl-2protein before and after irradiation were 73.3% and 46.7% respectively, with the difference beingsignificant (P 【 0.05); The positive rates of bax protein before and after irradiation were 86% and100% respectively, with the difference being significant (P 【 0.05). Conclusion: bax and Bcl-2protein play an important role in apoptosis induced by fractionated radiation therapy. Apoptosisinduced by irradiation is contributed to upregulation of bax protein or downregulation of Bcl-2protein.展开更多
基金supported by the National High Technology Research and Development Program of China(No. 2006AA02A107)the Major State Basic Research Program of China(Nos.2009CB521704 and 2010CB945504)the National Nature Science Foundation of China(No.30873031)
文摘MicroRNAs （miRNAs） are small, noncoding RNAs which can often act as an oncogene or a tumor suppressor. Several miRNAs are associated with the development of hepatoceUular carcinoma （HCC）. We demonstrated that miR-125b significantly suppresses HCC cell proliferation and promotes apoptosis by inhibiting the gene expression of the anti-apoptotic protein, Bcl-2. Bioinformatic analysis indicated that the 3tUTR of Bcl-2 has binding sites for miR-I25b, Luciferase reporter assay confirmed the ability of miR-125b to dramatically suppress Bcl-2 transcription, suggesting that Bcl-2 is a target gene for miR-125b. We concluded that miR-125b acts as a tumor suppressor in hepatic tumor development by targeting Bcl-2 and inducing cancer celt apoptosis.
基金supported by a grant from the Foundation of Most Advanced Group of Medical Scientists and Technicians of Shandong Province(2007GG30002014)
文摘BACKGROUND:Many Chinese herbs,especially herbal injections,have been shown to have anti-tumor effects in recent years.However,since most reports focus on the clinical effectiveness of these herbs,their mechanisms of action are not well understood.In this study,we assessed apoptosis in the hepatocellular carcinoma(HCC)cell line HepG2 induced by an injectable extract from the seed of Coix lacryma-jobi(Semen coicis,SC),and monitored the expression of Bcl-2 and caspase-8.METHODS:Injectable SC was applied to HepG2 cells at different concentrations and the cells were collected 12,24 and 48 hours later.5-fluorouracil was used as a positive control group,and fluorescence-activated cell-sorting cytometry was used to measure the apoptosis rate of HepG2 cells and the expression of Bcl-2 and caspase-8 proteins.RESULTS:SC induced apoptosis in HepG2 cells in a concentration and time-dependent manner,and the expression of caspase-8 was elevated and prolonged.However,it did not significantly influence the expression of Bcl-2.CONCLUSION:Injectable SC may induce apoptosis in HCC cells by regulating the expression of caspase-8.
文摘Objective:To investigate the relationship among Pokemon,NF-κB p65 and Bcl-2 in hepatoma cells.Methods:HCC cell HepG2,SMMC7721 and human fetal liver cell line L02 cells were used,and expression of Pokemon,NF-κB p65 and Bcl-2 in three cells were detected by realtime PCR and western blot.Then siRNA of Pokemon was applied to inhibit the expression of Pokemon and NF-κB p65 and apoptotic rate was determined by flow cytometric analysis. Results:Expressions of Pokemon,NF-κB p65 and Bcl-2 in human hepatoma cell HepG2, SMMC7721 expression were significantly higher than those in human embryonic stem cells L02. siRNA of Pokemon inhibited the expression of Pokemon,NF-κB p65 and Bcl-2 in liver cancer cells,and significantly increased apoptosis of liver cells.While siRNA of NF-κB n65 inhibited the expression of NF-κB p65 and Bcl-2,but Pokemon expression in hepatoma cells had no significant change.Conclusions:The proto-oncogene Pokemon can inhibit PI4ARF by specific transcription regulation of cell cycle and can induce tumors.In addition,Pokemon can regulate NF-κB p65 through the expression of apoptosis repressor,and promote the development of liver cancer.It suggests signal network in the liver include the regulation of new non-classical NF-κB regulatory pathway.
文摘Objective: The expression of B-cell lymphoma 2(Bcl-2) seems to be influenced by the endocrine environment. Numerous reports demonstrate the diverse expression of Bcl-2 family members under sex steroid regulation. With the exception of estrogen-related tumors, androgen-related tumors have shown their characteristics in Bcl-2 expression. In this study, the status of Bcl-2 expression in male hepatocellular carcinoma(HCC) patients was examined to verify the high incidence of HCC in males.Methods: Tumor tissue microarray was used to examine Bcl-2 expression levels in 374 HCC cases including 306 males and 68 females. Kaplan-Meier method, log-rank test, and Cox proportional hazards model were applied to investigate the predictive value of Bcl-2 in HCC patients.Results: Immunohistochemistry analysis showed that male patients with higher Bcl-2 levels had significantly longer median survival time and recurrence time than those with lower levels. However, no significant differences in outcomes were found between different Bcl-2 levels in female patients. When the male patients were stratified into several age points, the level of Bcl-2expression showed poorer predictive efficiency in the 45–49 and 55–60 age groups in andropause-age patients compared with other age groups. Bcl-2 was an independent prognostic factor for both overall survival(P < 0.0001) and recurrence time(P =0.0001) in male patients. After excluding male patients in the 45–60 age group, the predictive efficiency was enhanced(n = 147,OS, P = 0.0002, TTR, P < 0.0001).Conclusions: Bcl-2 expression is an independent predictor of survival and recurrence in male HCC. Bcl-2 levels may also be regulated by androgens or androgen receptors in male HCC patients. Bcl-2 levels change and exhibit poor predictive efficiency when androgen levels vary dramatically(andropause age).
文摘Objective: To correlate the frequency of p53 mutations, bcl-2 expression and the proliferation status (proliferating cell nuclear antigen, PCNA) in patients with bladder cancer with cell proliferation, apoptosis and their clinico-pathologic ?ndings. Methods: Para?n-embedded sections from 39 super?cial (T1G1-G3) and 23 invasive (T2-T4a G3 N0M0) primary transitional cell carcinomas (TCC) in the bladder were investigated immunohistochemically for p53, bcl-2 and PCNA. The median follow-up was 37 months; 24 had recurrences. The proliferation index (PI) was expressed as a percentage of the PCNA-positive cells in the tumor cells. Apoptosis was detected by terminal deoxy-nucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and the apoptotic index (AI) was expressed as a percentage of the TUNEL- positive tumor cells. Results: P53 mutation was identi?ed in 50 patients (80.6%). The mutation was most common in tumors of grade 3 (91.3%) as compared to grade 2 (78.5%) and grade 1 (72.7%, P<0.05). Stage pT2 tumors had a higher frequency of p53 mutation (95.7%) as compared to pTa-1 tumors (74.3%, P<0.01). Only 14 tumors (22.5%) expressed bcl-2; grade 3 tumors expressed bcl-2 signi?cantly more frequently (P<0.05); there was no correlation between bcl-2 and tumor stage. There was no interrelation between p53 mutation and bcl-2 expression (P>0.05). The PI ranged from 17.2% to 41.8% (median 22.4%) and the AI from 1.9% to 3.5% (median 2.9%) in bladder cancer. Statistical analyses revealed a close associations between PI, AI and tumor grade and stage of bladder cancer. Conclusion: P53 mutation correlates with invasion. P53 and PCNA overexpression may o?er valuable additional prognostic information in bladder tumors. With the progression of the tumor grade, cell proliferation may be accompanied by frequent apoptosis in bladder cancer, but the PI increased much more than the AI.
文摘Objective:To explore the influence of silencing Bcl-2 expression by small interfering RNA（siRNA） on Bcl-2 protein expression,cell apoptosis rale and radiosensilivity of gastric cancer BCC823 cells.Methods:siRNA segment for Bcl-2 gene was designed and synthesized,then was induced into gastric cancer BGC 823 cells by liposome transfection.Bcl-2 protein expression was detected by Western Blotting.After X radiation,flow cytometry and clone forming assay were used to determine the effects of RNA interference on BGC823 cell apoptosis rate and radiosensitivity. Result:After the transfection of Bcl-2 siRNA,the positive expression rate of Bcl-2 protein in BGC823 cells was（35.45±2.35）%.Compared with the control group and negative siRNA transfection group,the rate was significantly decreased（P【0.01）.The apoptosis rate of BGC823-RNAi cell was（10.81±0.91）%,which was significantly higher than the control group and negative siRNA transfection group（P【0.01）.After 48h X radiation,the apoptosis rate of BGC823-RNAi was（28.91±1.40）%,which was significantly higher than the control group and the group without radiation （P【0.01）.During clone forming assay D<sub>0</sub>,D<sub>4</sub> and SF<sub>2</sub> values in Bcl-2 siRNA1 transfection group were all lower than those in the control group.The radiosensitivity ratio was 1.28（the ratio of D<sub>0</sub>） and 1.60（the ratio of D<sub>4</sub>）.Conclusions:Specific siRNA of Bcl-2 gene can effectively inhibit the expression of Bcl-2 gene,enhance the radiosensitivity and apoptosis of gastric cancer BGC823 cells,having good clinical application perspective.
文摘Mitochondrial K＋-ATP （mito-KATP） channels play an important role in cellular function and survival following ischemic stress. The present results revealed that intervention with diazoxide, a mito-KATP channel opener, led to an increase in Bcl-2 expression in the cerebral cortex of rats subjected to cerebral ischemia reperfusion injury. In addition, the intervention also led to clear improvements in neuronal mitochondrial morphology and consciousness post-injury. Glibenclamide a mito-KATP channel blocker, exhibited the converse effects. Both diazoxide and glibenclamide exerted dose-dependent effects （in particular, at 18 mg/kg diazoxide and 25 mg/kg glibenclamide）. These findings suggest that diazoxide exerts a neuroprotective effect on cerebral ischemia reperfusion injury by opening mito-KATP channels and upregulating Bcl-2 expression.
文摘AIM: To examine the expression of thymidylate synthase (TS) and oncoprotein Bcl-2 in advanced colorectal cancer (CRC) patients,and to determine their mutual relationship,association to therapeutic response and impact on disease outcome.METHODS: Tumor samples from 67 patients with CRC,who were treated at advanced stage with either irinotecan alone or in combination with 5-fluorouracil/ leucovorin,were analyzed for expression of TS and Bcl-2 using immunohistochemistry.RESULTS: A significant linear correlation between lower expression levels of Bcl-2 and lower levels of TS expression was found (P = 0.033).Patients with high levels of both TS and Bcl-2 expression had a significantly longer disease-free survival (DFS) (42.6 mo vs 5.4 mo,n = 25) than those with low TS/Bcl-2 index (P = 0.001).Tumors with low levels of both TS and Bcl-2 were associated with a longer survival with metastasis (WMS) interval in the whole patients group (n = 67,P = 0.035).TS/Bcl-2 index was not significantly related to disease-specific survival.CONCLUSION: The present data suggest that CRC patients with low TS/Bcl-2 demonstrate a significantly shorter DFS and longer WMS.
基金the National Natural Science Foundation of China,No. 30770762
文摘We tested the effects of curcumin treatment on a rat model of Alzheimer＇s disease induced by beta-amlyoid （Aβ1-40） expression. We investigated alterations in the expression of the apoptosis-related genes Bax and Bcl-2 in the hippocampus, as well as changes in the spatial memory and cognitive function of the rats. Reverse transcription-polymerase chain reaction and immunohistochemistry results showed that Bax expression was remarkably decreased and Bcl-2 expression was increased in the rat Alzheimer＇s disease model after curcumin treatment. Morris water maze results showed that the average time of escape latency was shortened in the curcumin treated model rats. Our study shows that curcumin can significantly improve spatial learning and memory functions in rats with Aβ1-40-induced Alzheimer＇s disease by modulating Bax and Bcl-2 expression.
文摘Brain-derived neurotrophic factor was utilized in the present study to treat cell injury models induced by aggregated β-amyloid（25 35）. Methylthiazolyldiphenyl-tetrazolium bromide assay and western blot analysis showed that brain-derived neurotrophic factor provided neuroprotection against cellular apoptosis by suppressing the decline in β-amyloid（25 35）-induced cell activity and the increasing ratio of Bax/Bcl-2. After treating pheochromocytoma cells with tyrosine kinase receptor B receptor inhibitor K252a, brain-derived neurotrophic factor reverses the above- mentioned changes. The experimental findings suggested that brain-derived neurotrophic factor prevented β-amyloid peptide-induced cellular apoptosis by modulating Bax/Bcl-2 expression, and this effect was associated with binding to the specific tyrosine kinase receptor B receptor.
基金the Natural Science Foundation of Chongqing, No. 2005(54)
文摘The ventral tegmental area and the locus coeruleus are associated with psychological and physical dependence of opioid addiction. To date, very little is known about brain-derived neurotrophic factor （BDNF） and Bcl-2 gene and protein changes following morphine addiction. The present study utilized immunohistochemistry and in situ hybridization techniques, which revealed that there were increased BDNF levels, but decreased Bcl-2 levels in the prefrontal cortex, locus coeruleus, hippocampus, and the ventral tegmental area during morphine-dependence formation and abstinence. However, the levels of BDNF remained unchanged, and Bcl-2 expression was increased in the nucleus accumbens. These results showed that BDNF and Bcl-2 are involved in the development of morphine dependence, and precipitation of abstinence syndrome.
基金Supported by:the Natural Science Foundation of Chongqing,No. CSTC2006EB5030
文摘BACKGROUND： Estrogen has been clinically demonstrated to attenuate ischemic brain injury. However, the precise mechanisms remain controversial. OBJECTIVE： To investigate the effects of estradiol on angiopoietin-1 mRNA and Bcl-2 expression, as well as apoptosis and cerebral blood flow, in ovadectomized rats with focal cerebral ischemia following reperfusion. DESIGN, TIME AND SETTING： Randomized, controlled, animal experiment. The study was performed at the Central Laboratory, Chongqing Medical University from September to December 2005. MATERIALS： Estradiol benzoate was purchased from Shanghai Ninth Pharmaceutical Factory; corn oil was purchased from Walmart Supercenter; TUNEL kit, rabbit anti-rat Bcl-2 polyclonal antibody, and biotin-labeled goat anti-rabbit antibody were purchased from Wuhan Boster, China. METHODS： Healthy, female, 6-month-old Wistar rats-wild-type and estrogen alpha receptor gene knockout （ERKO）-were randomly divided into estradiol and control groups with 25 animals in each group. The rats were intramuscularly injected with estradiol benzoate （100 μg/kg per day） at 30 days following bilateral ovariectomy or corn oil （1 mL/kg per day） for seven consecutive days. Following administration, cerebral ischemia/reperfusion models were established using the right middle cerebral artery occlusion （MCAO） method. After 30 minutes of MCAO, estradiol and control groups were separately injected with estradiol benzoate and corn oil with the above-mentioned doses. MAIN OUTCOME MEASURES： Cell apoptosis was determined by TUNEL; angiopoietin-1 mRNA and Bcl-2 gene expression was determined, respectively, by immunohistochemical staining and RT-PCR. In addition, changes in cerebral blood flow were measured by laser Doppler flowmetry. RESULTS： Changes in angiopoietin-1 mRNA and cerebral blood flow in estradiol-treated, wild-type, MCAO rats following ischemia/reperfusion were greater than in control rats （P 〈 0.01 or 0.05）. However, no significant difference was observed between estradiol-treated ERKO MCAO rats and control rats. In addition, estradiol-treated wild-type and ERKO MCAO rats exhibited significantly increased Bcl-2 expression （P 〈 0.05） and decreased number of apoptotic cells in brain tissues compared with control groups （P 〈 0.05）. CONCLUSION： Estradiol upregulated angiopoietin-1 mRNA and Bcl-2 expression, suggesting that estradiol might be involved in protective mechanisms of cerebral ischemia/reperfusion injury.
文摘Macrophage cells play an important role in the initiation and regulation of the immune response. All-trans retinoic acid (ATRA) and its natural and synthetic analogs (retinoids) affect a large number of biological processes.Recently , retinoids have been shown promise in the therapy and prevention of various cancers. However, many interesting questions related to the activities of retinoids remain to be answered: (Ⅰ) Molecular mechanisms by which retinoids exert their effects; (Ⅱ) why the clinical uses of retinoids give undesirable side effects of varying severity with a higher freqllency of blood system symptoms; (Ⅲ)little is known for its impacts on macrophage cells etc. We set up this experiment, therefore, to examine the apoptosis of ATRA on macrophage Ana-1 cell line. Apoptosis of the cells was quantitated, after staining cells with propidium iodide (PI), by both accounting nuclear condensation and flow cytometry. When the cells were treated with ATRA at or higher than 1 μM for more than 24 h, significant amount of the apoptotic cells was observed. Induction of apoptosis of Ana-1 cells by ATRA was in time- and dose-dependent manners, exhibiting the similar pattern as the apoptosis induced by actinomycin D (ACTD). ATRA treatment of Ana-1 cells also caused the changes of the mRNA levels of apoptosis-associated gene bcl-2, as detected by Northern blot analysis. The temporal changes of bcl-2 expression by ATRA was also parallel to that by ACTD. In conclusion,ATRA can induce apoptosis in macrophage cells, which may be helpful in understanding of immunological functions retinoids.
基金Supported by Directive Subject of Chinese PLA(No.96L036)
文摘Objective: To investigate the relationship between oxidative stress and apoptosis and bcl-2 expression following traumatic brain injury (TBI). Methods: Male Sprague-Dawley rats were subjected to lateral fluid percussion brain injury (FPBI) of moderate severity. U-74389G (20 mg/kg) were administered intravenously before FPBI. The neurological functions were measured by beam-walk task (BWT) and beam-balance task (BBT). In addition to morphological evidence of apoptosis, TUNEL histochemistry was used to identify DNA fragmentation in situ with both light and electron microscopic levels. The internucleosomal fragments of DNA in apoptotic cells were examined using agarose gel electrophoresis. Bcl-2 protein expression was detected by immunohistochemistry. Results: The scores of BWT and BBT were significantly improved (P<0.01) in the treated animals. The treatment significantly reduced the number of apoptotic cells that was counted in the areas of the injured hemisphere at various time points following TBI. No DNA ladder was detected in the treated rats. Bcl-2 expression was observed in the cerebral cortex, subcortical white matter, dentate gyrus, hippocampal CA1 and CA3 region ipsilateral to injured hemisphere. Bcl-2 positive cells displayed normal nuclear morphology; Little Bcl-2 positive cells revealed morphological feature of apoptosis or necrosis. The immunoreactivity of Bcl-2 protein decreased significantly in the hippocampus ipsilateral impact site as early as 6 h post-injury. During 1-3 d after injury, the bcl-2 protein expression decreased relatively slow. In the U-74389G treated groups, the downregulation of bcl-2 expression was halted. Conclusion: In this model, apoptosis is associated with an activation of lipid peroxidation. U-74389G may block oxidative stress and halt the downregulation of bcl-2 expression. These may be one of the molecular mechanisms of the neuro-protective effects by U-74389G.
文摘Objective: To study the effect of auricular acupuncture on dysmnesia and the relationship between the memory improvement and bcl 2 protein expression in vascular dementia (VD) rats. Methods: Forty Wistar rats were randomized into control group, VD group, acupuncture+VD group and pseudo operation group, with 10 cases in each group. Rat VD model was established by using 4 vessel occlusion method. Otopoint "Nao" point and "Shen"(MA SC) were punctured, once daily continuously for 15 days. The rats’ memory capability was tested with Y maze method and bcl 2 expression of the brain tissues displayed by immunohistochemical method and measured using MIAS 2000 Image Analyzer. Results: Results showed that the scores of control group, VD group and acupuncture+VD group before operation were 5.68±1.29, 6.07±1.67 and 5.86±1.74 respectively, while following auricular acupuncture treatment, the scores of the 3 groups were 5.81±1.51, 18.06±2.68 and 8.31±1.85 separately, suggesting that the VD rat’s learning and memory abilities in acupuncture+VD group were raised apparently in comparison with those of VD group (P<0.01). In control, VD and acupuncture+VD group, bcl 2 immuno reaction positive neurons in CA 1 area of the hippocampus were 14.31±4.87, 28.67±5.63 and 65.74±8.19 respectively, displaying that the improvement of learning and memory abilities caused by auricular acupuncture treatment may be related to the up regulation of bcl 2 expression (an inhibitory gene of apoptosis). In comparison with control group, the loss of neurons in the pyramidal cell layer of the hippocampal CA 1 area of VD group was more severe, while that of acupuncture group was markedly lighter. Conclusion: Auricular acupuncture of otopoint "Nao" point and "Shen"(MA SC) can raise the learning and memory abilities of VD rats, which may be realized by its inhibitory effect on apoptosis and the protection action on ischemic hippocampal neurons.
文摘Objective: To study the expression of Fas and Bcl-2 proteins on T lymphocyte subsets in the peripheral blood of relapsing patients with condyloma acuminatum(CA) and healthy controls.Methods: Flow cytometry (permeabization and staining procedure with conjugated antibodies) was used.Results: We observed that the expression of Fas protein on CD4^+ T lymphocyte subset of CA patients was significantly higher than that of healthy controls( P<0.01 ).Conclusions: Increased expression of Fas proteinon CD4^+ T lymphocyte subset may be a cause of de-creased percentage of CD4^+ T lymphocyte subset. This induces the increased ratio of CD4^+/CD8^+.
基金This project was partially supported by Science Foundation of Lanzhou Command of PLA(No.YZ-0106).
文摘Objective: To evaluate the anti-tumor effects of SeO2 and its mechanisms on three human lung cancer cell lines. Methods: Three lung cancer cells A549, GLC-82 and PG were treated with 3-30 μmol/L SeO2. Flow cytometry was used to detect apoptosis, and analyze the changes of expression of p53 and Bcl-2, as well as ROS and Ca2+ level within cells. Results:SeO2 markedly inhibited cell proliferation and viability, and prompted apoptosis after 48 h treatment. SeO2 at 10 μmol/L induced 47.8% apoptosis in A549 cells, 40.8% in GLC-82 cells, 18.2% in PG cells. SeO2 at 30 μmol/L induced 37.8% apoposis in PG cells,but did not increase apoptotic raes in other two cells. SeO2 could down-regulate the mean fluorescent intensity of Bcl-2 from 65.8 to 9.6 in A549, but not in GLC-82 and in PG cells, up-regulate wild type p53 level in all three cells. SeO2 decreased the ROS and Ca2+ level markedly within three tested cells. Conclusion: SeO2 showed anti-tumor effect via apoptosis pathway in three lung cancer cell lines. The decrease of ROS and Ca2+ level within cells as well as regulation of Bcl-2 and p53 expression may play important roles in above apoptotic procedure.
文摘To evaluate the apoptosis positivity, the expression of Bcl-2, bax proteinsin 30 patients with squamous cell cervix carcinoma before and after radiotherapy. Methods: By usingimmuno-histochemical and TDT-dUTP nick end labelling techniques, 30 cases of squamous cell cervicalcarcinoma were analyzed. Results: The apoptosis positivity before and after irradiation was 76.7%and 100% respectively, with the difference being significant (P 【 0.05); The positive rates of Bcl-2protein before and after irradiation were 73.3% and 46.7% respectively, with the difference beingsignificant (P 【 0.05); The positive rates of bax protein before and after irradiation were 86% and100% respectively, with the difference being significant (P 【 0.05). Conclusion: bax and Bcl-2protein play an important role in apoptosis induced by fractionated radiation therapy. Apoptosisinduced by irradiation is contributed to upregulation of bax protein or downregulation of Bcl-2protein.
