Objective To study the expressions of Livin, Caspase-3 and Bcl-2 in lung tissue of nonsmall cell lung cancer ( NSCLC) , and their relationship with the clinicopathological features and prognosis of NSCLC. Methods The expressions of Livin, Caspase-3 and Bcl-2 proteins were evaluated by immunohistochemical method in 87 NSCLC samples and 40 lung benign tissues. The relationship of their expressions with the clinicopathological features and prognosis of NSCLC were analyzed by Spearman’s Rank correlation and COX Regression. Results More NSCLC tissues showed expression of Livin than lung benign tissues( 72. 41% vs 0. 0% , P = 0. 000 ) , and the expression of Caspase-3 was significantly decreased ( 67. 82% vs 87. 5%, P lt; 0. 05 ) . The proteins of Livin, Caspase-3 and Bcl-2 were detected in the endochylema but none was detected in nucelus. There was no relationship between the expression of each of these proteins and the clinicopathological features of NSCLC such as histologic type, tumor differentiation,lymph node metastasis, TNM stage, the size of tumor, and tumor site. The expression of Livin was correlated with Caspase-3 and Bcl-2 expressions ( r1 = - 0. 260, P = 0. 015; r2 = 0. 351, P = 0. 001) . Livin, Caspase-3 and Bcl-2 were not independent prognostic factors of NSCLC. Conclusions The expression of Livin and Bcl-2 are up-regulated in NSCLC. The expression of Livin is positively correlated with that of Caspase-3 and Bcl-2, they might interact with each other in the carcinogenesis and development of NSCLC. The levels of Livin, BCl-2 and Caspase-3 proteins are not independent factors affecting the prognosis of lung cancer patients.
Abstract: Objective To investigate the expression of inhibitor of apoptosis gene Livin and its relationship with expression of P53,Bcl-2 in esophageal carcinoma tissues. Methods The expression of Livin messenger ribonucleic acid (mRNA) in 36 esophageal carcinoma tissues and 18 paracancerous tissues were measured by reverse transcriptionpolymerase chain reaction (RT-PCR) combined with silver staining technique. The expression of Livin, P53 and Bcl-2 proteins were detected by immunohistochemical method (streptavidin-peroxidase). Results RT-PCR results: Livin mRNA positive expression of esophageal carcinoma tissues was more evident than that of paracancerous tissues, the expression of both variants was simultaneous basically. Immunohistochemical results: the Livin protein positive expression rate of esophageal carcinoma tissues was higher evidently than that of paracancerous tissues(Plt;0.01). Livin protein positive expression rate of external coat of esophagus invaded by carcinoma was higher than that of tunica muscularis esophagi invaded by carcinoma(Plt;0.05); Livin protein positive expression rate of lymph node metastasis was higher than that of normal lymph node (Plt;0.05). The expression of Livin protein was not related to the expression of P53 protein（χ2=1.00,P=0.505）,but it was positively related to the expression of Bcl-2 protein（χ2=10.60，P=0.003）. Conclusion Aberrant expression of Livin may be a new target for diagnosis and gene treatment of esophageal carcinoma.The aberrant expression of Livinand apoptosis related gene Bcl-2 may play synergetic roles in process of carcinogenesis of esophageal carcinoma.
Objective To investigate the expressions of heat shock protein 27 (HSP27), Bcl-2, and Bax proteins of the nerve cells after spinal cord ischemia/reperfusion injury (SCII) in rats and their relationship. Methods Seventy adult male Sprague Dawley rats (weighing, 200-220 g) were randomly divided into the sham operated group (sham group, n=35) and the SCII group (n=35). Only the left renal artery was exposed with no occlusion of the abdominal aorta in the rats of sham group. The left renal artery was exposed with occlusion of the abdominal aorta for 20 minutes in the rats of SCII group. At 4, 8, and 12 hours and at 1, 2, 3, and 5 days, reperfusion treatment was performed in 5 rats respectively, and then the spinal cord tissue was harvested to detect the expressions of HSP27, Bcl-2, and Bax protein of the nerve cells by using immunohistochemistry staining. Results The HSP27 began to express at 4 hours, reached the peak at 3 days, and decreased at 5 days in SCII group; significant differences were found between at 3 and 5 days and at the other time points (P lt; 0.05). The Bcl-2 expression increased at 4 hours, reached the peak at 1 day and maintained a high level at 2 days, and then gradually decreased; significant differences were found between at 1 and 2 days and at the other time points (P lt; 0.05). The Bax expression reached the peak at 12 hours and 3 days, and decreased at 5 days; significant differences were found between at 12 hours and 3 days and at the other time points (P lt; 0.05). A little expression of each protein was observed in sham group at different time points; the expressions of HSP27, Bcl-2, and Bax proteins in SCII group were significantly higher than those in sham group at different time points (P lt; 0.05). Conclusion There may be the time window of self repair after SCII. High expression of HSP27 has an obvious protective effect on the SCII in rat, by promoting the expression of the anti-apoptotic protein Bcl-2 and reducing the expression of the pro-apoptotic protein Bax so as to inhibit spinal cord cell apoptosis.
【Abstract】 Objective To detect the expression of Bcl-2/adenovirus E1B 19-kDa-interacting protein 3 (BNIP3)in cell death induced by nutrition deprivation in nucleus pulposus cells so as to further understand the mechanism of deathin nucleus pulposus cells. Methods Two adult Sprague Dawley rats, male or female, weighing 150-200 g, were involvedin this experiment. The cells isolated from rat caudal disc were cultured under the condition of L-DMEM culture media,10%FBS, and 21%O2 (control group) and under the condition of DMEM-free glucose culture media, no serum, and 1% O2(experimental group). The expressions of BNIP3 gene and protein were detected by real-time fluorescent quantitative PCR,immunofluorescence staining, and Western blot. The cell apoptosis rate and mitochondrial membrane potential were measuredby flow cytometry at 24, 48, and 72 hours after culture. Results The expression of BNIP3 decreased in the control group;the expressions of BNIP3 showed an increasing tendency with time in the experimental group, and BNIP3 combined withmitochondria. Significant differences were observed in the expressions of BNIP3 gene and protein between 2 groups at the othertime (P lt; 0.05) except that no significant difference was observed in the expression of BNIP3 gene at 24 hours (P gt; 0.05). Thecell apoptosis rate and mitochondrial membrane potential were significantly lower in the experimental group than those in thecontrol group (P lt; 0.05). Conclusion Upregulation of BNIP3 and translocation to mitochondria may be involved in nucleuspulposus cell death in nutrition deprivation.
Objective To investigate the expression of Bcl-2 in acute leukemia of different pathological states and its relationship with chemotherapeutic efficacy. Methods Case-control studies and cohort studies were collected by searching the electronic bibliographic databases such as CBMdisc (1979 to 2010), Chinese Sic-tech Periodical Full-text Database (1989 to 2010), WanFang (1982 to 2010), Chinese Journals Full-text Database (since 1994), China Master’s Theses Full-text Database (since 1999), and China Doctor Dissertations Full-text Database (since 1999). All the relevant studies were identified and the quality of the included studies was assessed. The RevMan 5.0 software was used for meta-analysis. Results A total of 10 studies were included. The results of meta analyses showed: the complete remission of acute leukemia with Bcl-2 positivity was lower than that of the Bcl-2 negative patients after chemotherapy and the difference between them was significant (OR=0.26, 95%CI 0.14 to 0.46); the difference between acute lymphocytic leukemia and acute non-lymphocytic leukemia in terms of Bcl-2 positive rate was not significant (OR=0.87, 95%CI 0.46 to 1.65); the Bcl-2 positive rate in complete remission (CR) patients after chemotherapy was significantly lower than that of partial remission (PR) and none remission (NR) patients (SMD= –0.87, 95%CI –1.53 to –0.20, P=0.01). Conclution The current domestic evidence proves that Bcl-2 is significantly correlated with the remission rate of acute leukemia patients, but more high-quality studies are still needed.
【Abstract】ObjectiveTo investigate the protective effects and the impact on the expression of Bcl-2 and Caspase-3 mRNA by Panax notoginseng saponins (PNS) preconditioning in rat liver transplantation. MethodsMale Sprague-Dawley rats were used as donors and recipients of orthotopic liver transplantation (OLT) and were divided into PNS preconditioning group (PNS group) and NS control group (NS group) randomly according to whether PNS was injected by venous (50 mg/kg) 1 h before liver grafts harvesting. The animals were respectively killed 2 h, 6 h and 24 h after reperfusion. Plasma samples were collected for ALT and AST test. Liver tissues were collected to detect histological changes, apoptosis and the expression of Bcl-2, Caspase-3 mRNA. ResultsThe serum levels of ALT and AST and the apoptosis index (AI) of liver tissue in PNS group were lower than those in NS group’s significantly (P＜0.05) at 2 h, 6 h and 24 h after reperfusion. The expression of Bcl-2 mRNA was enhanced significantly in PNS group at 6 h, 24 h after reperfusion and the expression of Caspase-3 mRNA was decreased significantly in PNS group at 2 h, 6 h after reperfusion as compared with NS group’s(P＜0.05). ConclusionPNS preconditioning can attenuate liver grafts ischemia/reperfusion injury and apoptosis of hepatocytes. Affecting expression of Bcl-2 and Caspase-3 genes may be one of the mechanisms of PNS antiapoptotic effects.
ObjectiveTo investigate expressions of ALCAM/CD166, Bcl-2, and Ki-67 in breast infiltrative ductal cancer tissues, so as to assess the role of ALCAM/CD166 protein in the carcinogenesis and progression of breast infiltrative ductal cancer. MethodsThe expressions of ALCAM/CD166, Bcl-2, and Ki-67 proteins were examined by immunohistochemistry(ElivisionTM Plus) in 96 breast infiltrative ductal cancer specimens and 30 adjacent normal tissues of breast cancer specimens(control group). The relation between ALCAM/CD166 protein expression and patient's age, tumor diameter, histopathologic grade, axillary lymph node metastasis, or TNM stage of breast infiltrative ductal cancer was analyzed, and the correlation between ALCAM/CD166 expression and Bcl-2 or Ki-67 was analyzed too. Results①In 96 cases of breast infiltrative ductal cancer, the positive rate of ALCAM/CD166 protein expression was 79.2%(76/96), which was significantly higher than that in the control group〔10.0%(3/30), P < 0.01〕.②In the breast infiltrative ductal cancer tissues, the expression of ALCAM/CD166 was related to axillary lymph node metastasis(P < 0.05), but was not related to patient's age, tumor diameter, histopathologic grade, and TNM stage(P > 0.05).③The ALCAM/CD166 protein expression was positively related to Bcl-2(rs=0.307, P=0.001) and not related to Ki-67(rs=0.064, P=0.475). ConclusionALCAM/CD166 protein expression might be related to the apoptosis and metastasis of breast infiltrative ductal cancer cells and it could serve as an important marker for predicting biological behavior and prognosis of tumor.
ObjectiveTo investigate the protective effect of Roux-en-Y gastric bypass surgery on early damage of renal tissue in type 2 diabetes mellitus rats, and explore the mechanism of the protective effects. MethodsDiabetes mellitus animal models were induced by intraperitoneal injection of streptozotocin (STZ, 35 mg /kg) and a high-fat diet.Diabetic rats were divided into three groups randomly (digital table method): diabetes control group (n=8), sham operation group (n=8), and Roux-en-Y gastric bypass group (n=14).Another 8 normal SD rats as the normal control group.The fasting blood glucose, serum total cholesterol (TC), triglyceride (TG), and free fatty acid (FFA) were measured before operation and in 8 weeks after operation; plasma BUN and Cr were measured respectively before operation and in 4 and 8 weeks after operation in each group rats, 24 h urine microalbumin and urine 8-hydroxydeoxyguanosine were measured respectively before operation and in 8 weeks after operation in each group rats.Renal pathological changes were observed and the indexes of kidney hypertrophy, the mean glomerular area (MGA), and the mean glomerular volume (MGV) were analyzed in 8 weeks after operation.The expressions of fibronectin, typeⅣcollagen (CoⅣ), transforming growth factor-β1 (TGF-β1), intercellular adhesion molecule-1(ICAM-1), nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4), and Bcl-2 protein in renal tissues were investigated by immunohistochemical staining. ResultsRoux-en-Y gastric bypass surgery could reduce the blood glucose, blood lipid, MGA, MGV, and the index of kidney hypertrophy of diabetic rats significantly (P < 0.05), improved renal pathological morphology and kidney function (P < 0.05), reduced the protein expressions of fibronectin and CoⅣ, decreased the protein expressions of TGF-β1, ICAM-1, and NOX4, and increased the protein expression of Bcl-2. ConclusionRoux-en-Y gastric bypass surgery can improve kidney function and the pathological damage of diabetes rats, its mechanism may be related to inhibition the protein expressions of TGF-β1, ICAM-1, and NOX4, and increase the protein expression of Bcl-2.
Objective To observe the structural changes of urinary center and the expression of Bcl-2 after conus medullaris injury in rats brain so as to explore the possible influence factors of degeneration in brain. Methods Thirty-six adult Sprague-Dawley rats were randomly divided into experimental group (n=30) and control group (n=6). In the experimental group, the conus medullaris injury model was established by cutting off the spinal nerve below L4, and no treatment was done in the control group. The modeling operations in the experimental group were successful, and 2 rats died at 3 months and 5 months after modeling operation respectively, which may be caused by renal failure or urinary tract infection. In the experimental group, 6, 6, 6, 5, and 5 rats were killed at 1 day, 1 week, and 1, 3, 6 months after operation respectively, and 1 rat was killed at each time point in the control group. The dorsolateral tissue of the pontine tegmentum was harvested to perform HE staining and Bcl-2 immunohistochemical SP staining. Results HE staining showed that there was no obvious difference between the experimental group and the control group at 1 day after operation, the neurons were densely packed, arranged neatly, and the nucleoli were clear; at 1 week, the space between the neurons in the experimental group were slightly widened; at 1 month, nucleus retraction in some neurons happened in the experimental group; at 3 and 6 months, the nuclei in the experimental group were more and more condensed, and even some cells disappeared. Bcl-2 immunohistochemical SP staining showed that the expression of Bcl-2 in the control group was weakly positive. The positive expression of Bcl-2 was found at 1 day after operation in the experimental group; the positive expression of Bcl-2 at 7 days after operation was significantly higher than that in the control group, and reached the peak; the positive expression of Bcl-2 decreased gradually at 1, 3, and 6 months after modeling operation, but it was still higher than that of the control group. Conclusion The urinary center appears structure degeneration and necrocytosis after conus medullaris injury in rats brain. The elevated expression of Bcl-2 may be associated with brain tissue repair and function remodeling.
ObjectiveTo detect the expression of Notch1, Bax, Bcl-2 genes in rat knee joint cartilage cells in a state of activation and inactivation of the Notch signaling pathway, and preliminarily study the mechanism of Notch signaling pathway on experimental rat knee osteoarthritis (OA) chondrocytes apoptosis.MethodsA total of 34 specefic-pathogen-free Sprague Dawley rats were selected, of which 32 were established the right knee OA models using Hulth method, and the other 2 were normally fed. Four weeks later, two randomly selected OA rats and the two normally fed rats were put to death, to observe the morphological changes of the right knee and ensure the OA models were successfully established by pathology examination. The remaining 30 rats were randomly divided into three groups with 10 in each. The rats were injected intra-articularly on each Tuesday and Friday, with Nocth signal pathway specific activator Jagged1 protein (25 ng/kg) in the activation group, γ-secretase inhibitor DAPT (GSI-IX) (100 ng/kg) in the inhibition group, and phosphate-buffered saline in the control group, respectively. The rats were sacrificed after 8 weeks of articular cavity injection. Taking the right knee articular cartilage speciments of femoral condyle, we observed the degeneration of articular cartilage of the three groups, observed the histomorphological changes by microscope, evaluated the Mankin scores, and used the immunohistochemistry to detect the expression of Notch1, Bax, Bcl-2 proteins.ResultsAfter the 8-week articular cavity injection, the Mankin scores in the activation group, the inhibition group, and the control group were 3.40±0.84, 6.70±0.95, 11.10±1.37, respectively, and the differences between the three groups were statistically significant (P<0.05). The positive rates of Notch1 and Bax of chondrocyte in the inhibition group were lower than those in the control group and the activation group (P<0.05), while the positive rate of Bcl-2 of chondrocyte in the inhibition group was higher than that in the control group and the activation group (P<0.05).ConclusionActivating the Notch signaling pathway may facilitate the chondrocyte apoptosis and aggravate OA by up-regulating Bax protein expression and down-regulating Bcl-2 protein expression; inhibiting the Notch signaling pathway may inhibit the chondrocyte apoptosis and relieve OA by up-regulating Bcl-2 protein expression and down-regulating Bax protein expression.