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Objective To investigate the expression and significance of myeloid cell leukemia-1 (Mcl-1 )gene and protein in Hepatocellular Carcinoma(HCC).Methods The expression of Mcl-1 was detected respectively by Reverse Transcription-Polymerase Chain Reaction (RT-PCR),Western blot and ENVISION immunohistochemistry in 20 HCC specimens,19 liver cirrhosis(LC)specimens,and 12 control ones.Their relationship with clinical and pathological characteristics of HCC was investigated.Results The expression of Mcl-1 mRNA in the control group,LC group and HCC group was 0.52±0.17, 3.46±1.7,3.65±2.93,respectively.The level in HCC and LC group was statistically different compared with the control group,respectively (t=7.925,5.334,P<0.05).The relative expression of Mcl-1 protein in LC group (0.51±0.35)and HCC group (0.75±0.36)were significantly higher than that in the control group (0.21±0.19)(t=5.526,6.355,P<0.05).The positive expression rate of Mcl-1 in HCC group was 55.00% (11/20),significantly higher than that in the con-trol group 33.33% (4/12)(t=7.835,P<0.05).The positive expression of Mcl-1 was related to tumor necrosis and TNM staging (χ2=4.201,P<0.05).Conclusion Mcl-1 gene and protein are differentially expressed in HCC,LC and the control, which may be involved in the occurrence,development and malignant transformation of HCC.
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Background:Secondary lymphoid tissue chemokine( SLC)is involved in lymphoid homing and anti-tumor immune response,and has a chemotactic effect on intestinal lymphocytes. Several animal studies have shown that SLC is involved in the occurrence and development of ulcerative colitis( UC). Aims:To investigate the expression and significance of SLC in UC. Methods:Forty active UC patients from Dec. 2010 to Dec. 2012 at Affiliated Hospital of Nantong University were enrolled,and 20 healthy volunteers were served as controls. Expression of SLC in the colon mucosa was detected by immunohistochemistry,and its relationship with severity of UC was analyzed. Results:SLC was positively expressed in all UC patients,while was negatively or weakly positively expressed in controls. Expression of SLC in UC patients was significantly higher than that in controls(4. 16 ± 0. 78 vs. 0. 52 ± 0. 11,P<0. 05). Expression of SLC was correlated with the severity of involvement of UC. Conclusions:Expression of SLC participates in development and progress of UC. SLC may play an important role in the induction of local damage and pathological changes of UC.
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Objective To investigate the dynamic expressions of CXCL11 and its role in the pathogenesis of acute necrotizing pancreatitis (ANP).Methods Forty-eight SD rats were randomly divided into control group and ANP group,with 24 rats in each group.ANP model was induced by retrograde injection of 4％ sodium taurocholate (1 ml/kg body weight) into the biliary and pancreatic duct.The rats were sacrificed at 1,3,6,12 hours.Serum level of amylase was determined,pathological changes in pancreatic tissue were routinely observed and scored.The expression of CXCL11 mRNA and proteon in pancreas was measured by fluorescence quantitative polymerase chain reaction and immunohistochemical method.The serum levels of CXCL11 were measured by enzyme-linked immunoadsorbent assay.Results The serum levels of amylase in ANP rats were significantly higher than those in control group [(6153 ± 355)U/L vs (185 ± 32)U/L at 6 h,P ＜0.05],pathological changes in pancreatre tisues were more significant in ANP rats,and the pathological score was significantly higher than that in control group [(9.00 ± 0.63) vs (0.33 ± 0.12) points at 6 h,P ＜ 0.05] ; the expressions of CXCL11 mRNA and protein in pancreatic tissue were significantly increased than those in control group (3.13 ± 0.43 vs 0.99 ± 0.24,2.76 ± 0.27 vs 0.33 ± 0.12 at 6 h,P ＜ 0.05).The serum level of CXCL11 was significantly higher than that in control group [(112.1 ± 14.2)ng/L vs (56.8 ±4.3) ng/L at 6 h,P ＜0.05)].Conclusions CXCL11 is an early inflammatory mediator in acute pancreatitis,and involved in the pathogenesis of ANP in rats.
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Objective To investigate the expression of galectin-3 in pancreatic cancer cell and its effect on the proliferation and invasion of SW1990.Methods Immunocytochemistry and semi-quantitative RT-PCR was used to detect the expression of galectin-3 protein and mRNA in SW1990,PANC1 and ASPC-1 cell lines.Galectin-3 mono-antibody of different concentrations ( 1,2,3,5 μg/ml) was used to treat SW1990 cells for 24,48,72 h,CCK-8 kits were used to detect the proliferation in SW1990 cells; Transwell chamber was used to study the invasion in SW1990.Results Expression of galectin-3 protein and mRNA was present in SW1990,PANC1,and ASPC-1.Galectin 3 mono-antibody inhibited the proliferation and number of invasive cells in a dose and time dependant manner.The inhibitory rates at 72 h were 19.8％,29.9％ and 42.7％ in 2,3,5 μg/ml galectin 3 mono-antibody groups,the difference among them and control group was statistically significant ( P ＜ 0.05 ).The inhibite rate of permeating membrane cells in 3 μg/ml galectin-3 mono antibody was 37.1％,the difference between this group and control group was statistically significant (P ＜0.05 ).ConcLusions Galectin-3 is highly expressed in pancreatic cancer cells.Galectin-3 antibody can inhibit the proliferation and migration capability of SW1990 cells.
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Objective To investigate the effects of two inhibitors of arachidonic acid metabolic pathway (5-cyclooxygenase blockade MK886 and COX 2 blockade celecoxib) on growth and VEGF mRNA expression of human pancreatic cancer cell SW1990.MethodsPancreatic cancer cells SW1990 were cultured with different concentrations of MK886,celecoxib,MK886 and celecoxib,then the cell proliferation was detected by using CCK-8,BLT1 mRNA,PGE2 mRNA and VEGF mRNA expressions were determined by RTPCR.ResultsAfter 10 μmol/L MK886 or 20 mmol/L celecoxib treatment for 24 h,the growth of SW1990 was greatly suppressed ( 1.80 ±0.06 vs 1.65 ±0.10,2.04 ±0.03 vs 1.86 ±0.02,P ＜0.01 ),and the growth suppression of SW1990 cells was increased accompanying the raised concentration of MK886 or celecoxib.After both MK886 and celecoxib treatment for 12 h,the growth of SW 1990 cells was much obviously suppressed (1.72 ±0.05 vs 1.52 ±0.05,P ＜0.01 ).After celecoxib treatment for 48 h,the BLT1 mRNA,PGE2 mRNA and VEGFmRNA expressions were not significantly changed,but the expressions of PGE2 mRNA were significantly decreased ( P ＜ 0.05 ).After MK886 or MK886 + celecoxib treatment,the expressions of BLT1 mRNA,VEGF mRNA were significantly decreased ( P ＜ 0.05 ),but the expressions of PGE2 mRNA were not significantly changed when compared to control group.ConclusionsTwo metabolic pathways of arachidonic acid have a close relation with occurrence and proliferation of pancreatic cancer,when both of the pathways were blocked,the proliferation of the pancreatic cancer cell was suppressed obviously.
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Objective To investigate the relationship between expression of nuclear factor kappa B p65 ( NF-κB p65) and hippocampal neuronal apoptosis in acute necrotizing pancreatitis (ANP) rats with brain injury. Methods Sixty-four SD rats were randomized into normal saline group (NS) and ANP group. The ANP rat model was induced by retrograde injection of 4% sodium taurocholate into the pancreaticobiliary duct of SD rats. Nissle stain was used to detect the brain injury. Neuronal apoptosis was determined by TUNEL.NF-κB p65 expression was detected by immunohistochemistry and RT-PCR. Results Hippocampal neuron was absent, karyopyknosis, unclear nucleolus and decreased Nissl bodies were found, the injuries was aggravated with time. The apoptosis index at the 3, 6 and 12 h in ANP group was 10.63 ±0.24, 21.02±0.25, 17.12±0.36, respectively, while they were 0.33±0.19,0.71±0.67, 0.45 ± 0. 33 in NS group, and the difference was statistically significant ( P ＜ 0. 01 ). The expressions of NF-κB p65 mRNA were 0. 63 ± 0.05,1.05 ±0.06,0.92 ±0.05, which were significantly higher than those in the NS group (0.11 ±0.01,0.12±0.01,0.08±0.01,P＜0.05).The chatge of expression of NF±κB p65 protein was consistent with that of NF-κB p65 mRNA. Conclusions The brain injury of ANP rats was highly correlated with neuronal apoptosis at the early and middle phase of ANP, and its mechanism may be related with NF-κB p65 activation.
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Objective To investigate the role of CC chemokine ligand 20 (CCL20) and CC chemokine receptor 6 (CCR6) in the pathogenesis of acute necrotizing pancreatitis (ANP). Methods 48 SD rats were randomly divided into two groups: control group and ANP group. The ANP model was induced by retrograde infusion of 4 % sodium taurocholate into the biliary and pancreatic duct in SD rats. The same amount of saline was injected in the control group. The rats were sacrificed at 1, 3, 6, 12 h, the serum amylase levels and the pathological score of the pancreas were measured. The expressions of CCL20 and CCR6 mRNA and protein in pancreas were detected by immunohistochemistry and semi-quantitative RT-PCR,respectively. Results The levels of serum amylase and the histological score of ANP group were significantly higher than those of control group (P ＜ 0.01 ). The expression of pancreatic CCL20 mRNA and protein was increased in a time-dependant manner ( P ＜ 0.05 ). The expression of pancreatic CCR6 mRNA at 6h was significantly higher than that of control group (0.88 ± 0.05 vs 0. 23 ± 0.09, P ＜ 0.01 ). The expression of pancreatic CCR6 mRNA at 12h was decreased when compared with that of 6h group, but it was still higher than that of control group (0.37 ± 0. 10 vs 0. 15 ± 0.07, P ＜ 0.05 ), the change of CCR6 protein was consistent with that of CCR6 mRNA. Conclusions CCL20 and CCR6 may play an important role in the pathogenesis of ANP.
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Objective To study the efficacy of RNA interference targeting 5-LOX on 5-LOX and VEGF expression as well as the growth of tumor xenografts in the nude mice,in order to evaluate the value of the clinical application.Methods SW1990 cells were injected into the back of BALB/c nude mice.Once the visible tumors were evidenced about 100mm3,the animals were divided randomly into 4 groups (6 animals/group) and treated with shRNA1 and shRNA2 targeting 5-LOX,negatrve control shRNA (shNC) or control LipofectamineTM 2000 (Lipo) by intratumoral injection.Observing the effect of the shRNA on the growth of tumor xenografts,investigating the expression of 5-LOX and VEGF by immunohistochemistry and RT-PCR.Results Two nude mice were dead in shNC group and Lipo group because of the wasting disease.Other nude mice had no changes in body weight,spirit,appetite,and activity.The growth of tumor xenografts was suppressed potently when administered with shRNA.Compared with shNC and Lipo group,the mean tumor size in groups treated with shRNA was reduced markedly at every point of test time.Between two treat groups,they did not have significant difference.5-LOX and VEGF were both expressed in the pancreatic cancer tissue.The level of the 5-LOX expression in shRNA groups was stronger than that in shNC or Lipo group.The VEGF had the game situation.Between the two treat groups,the difference was not significant.Conclusions RNA interference targeting 5-LOX can inhibit the growth of tumor tumor xenografts in the nude mice by depressing the expression of 5-LOX directly and depressing the expression of VEGF indirectly.
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Objective To investigate the effects of inhibition of 5-LOX by RNA interference on proliferation suppression and apoptosis induction of pancreatic cancer cell line. Methods Plasmid expression vectors containing four 5-LOX siRNA array and one negative control array were established, resoectively, and were transfected into pancreatic cancer cell line SW1990 with Lipofectamine TM2000. Cell proliferation inhibition rate was measured by MTT assay; apoptotic rate was examined by flow cytometry. Results The inhibitory rate of expression of 5-LOX mRNA in negative control group and four 5-LOX siRNA groups was (3.0 ±1.4)%, (18.8±1.5)%, (53.5±2.3)%, (56.1±2.0)%, (52.4±2.5)%; the inhibitory rate of expression of 5-LOX protein was (4.5 ± 2.0) %, (18.1 ± 2.5) %, (50.4 ± 4.3) %, (48.9 ± 4.4) %, (45.9 ± 4.0) %. The inhibitory rates of cancer cell proliferation at 24 h and 48 h after the transfection were (2.1±1.0)%, (5.5±1.3)%, (11.9±1.2)%, (13.4±1.1)%, (13.8±1.3)% and (3.0±1.3)%, (16.0 ± 2.2) %, (25.7 ± 2.5) %, (25.3 ± 3.1) %, (27.2 ± 3.2) %, respectively. The apoptotic rates at 48 h after the transfection were (3.0 ± 1.0) %, (7.1 ± 1.10%, (17.5 ± 0. 9) %, (21.5 ± 1.1) %, (15.7 ± 1. 0)%, respectively. Conclusions The plasmid vector containing siRNA against 5-LOX could suppress 5-LOX expression in SW1990 cells effectively and specifically, and could inhibit proliferation and induce the apoptosis of pancreatic cancer cells.
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Objective To investigate the suppression effects of Tripotolide (TL) on the pancreatic cancer xenograft models and angiogenesis. Methods The growth suppression effect of TL on SW1990 was determined using cell count kit (CCK-8), apoptotic cells induced by TL were examined by morphology and terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay. The inhibitory effects of TL on the growth of tumor xenografts and tumor microvascular density (MVD) were investigated. ResultsTL inhibited the growth and proliferation of SW1990 cells in a concentration-dependent and time-dependent manner. The inhibition ratios of cells treated at 160 mg/ml TL for 24 h was 50. 6%, the apoptotic rate increased from 9.6% in the control group to 45.1% (P <0.01 ). The inhibition rate of cancer xenograft growth was 89.9% when TL was intratumorally injected at the dose of 0.5 mg/kg. The expression of VEGF in tumor tissue decreased while MVD also decreased from 36.25±8.64 to 9.87±3.34 (P <0.01 ). ConclusionsTL induced prominent growth inhibition and apoptosis in human pancreatic cancer cell lines. TL.can attenuate the growth of pancreatic caner xenografts through its effect on antiangiogenesis.
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Objective To investigate the 5-lipoxygenase(5-LOX)expression in human pancreatic cancer cells and the inhibitory effect on growth of SW1990 by selective 5-LOX inhibitor zileuton in vitro.Methods The 5-LOX expression in pancreatic cancer cell was defected by immunocytochemistry and RT-PCR form May 10,2003 to May 25,2004.MTT and flow cytometry with AnnexinⅤ-FITC and propidium iodide(PI)staining and acridine orange(AO)staining were used to observe the effect of zileuton on the growth of SW1990cell line.Results The 5-LOX mRNA and protein expression was detectable in pancreatic cancer cell line by mmunohistochemistry and RT-PCR technique respectively.Zileuton could inhibit the growth of SW1990 cell line in a time-and dose-dependent manner.Conclusion Zileuton might inhibit the growth of SW1990 cell line in a time-and dose-dependent manner.
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Objective To evaluate the usefulness of cytologic examinations in the diagnosis of pancreatic neoplasms using nasopancreatic drainage and pancreatic duct brushing during ERCP.Methods In 47 patients with pancreatic diseases,cytologic examinations of pancreatic duct brushing and nasopancreatic drainage during ERCP were performed.Results The rate of accuracy,sensitivity,and specificity of brush cytologic examination were 70^4%,65^2% and 100%,respectively,and that of pancreatic juice cytologic examinations was 45%,8^3% and 100%,respectively.Conclusion Our results confirm the value,safety,and utility of obtaining cytologic specimens at the time of ERCP for the early diagnosis of pancreatic neoplasms.
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Objective To evaluate the usefulness of cytologic examinations of pancreatic duct brushing obstained during ERCP in diagnosis of pancreatic neoplasms. Methods In 27 patients with suspected manlignancy, cytologic examinations of pancreatic duct brushing during ERCP were performed. Results The positive rate of brush cytology was 55.6%.The results were affected by the location of lesion, correct cytodiagnoses of cancer were 69.2% in cancer at the head of the pancreas and 60% in cancer of the body. The accuracy rate of ERCP was 77.8%.However, by combining these 2 methods, the accuracy rate rose to 100%. Conclusion Our results confirm the significance,safety,and usefulness of the cytologic specimens obtained at the time of ERCP for the diagnosis of pancreatic neoplasms.