Effect of nuclear factor-κB p65 ASOND on I typ collagen expression and rat hepatic stellate cells proliferation

Objective Sstudy effect of nuclear factor-κB ASOND on I type collagen expression and rat hepatic stellate cells(HSC)proliferation.Methods Rat HSCs were separated by affusing and digestingof Ⅳ type collagenenzyme and density acentric method.Lipid-mediated NF-κB p65 ASOND(0.001,0.01,0.1,1μmol/L)Was transferred into rat HSCs.Toxicity of HSCs caused by NF-κB p65 ASOND and activity of LDH were determined by trypan blue staining.Proliferation affection of transferring NF-κB p65 ASOND into HSCs was determined by MTT.In different concentration NF-κB p65 ASOND.expression of Ⅰ type collagen stimulated by 1mg/L TNF-αwas determined by RT-PCR and ELISA.Results After transfection of NF-κB p65 ASODN,expression of NF-κB protein in HSCs was decreasing.Toxicity experiment indicated that NF-κB p65 ASOND of different concentration(0.001,0.01,0.1 and 1.0 μmol/L)had no effect on HSCs livability and LDH activity(P<0.05).Four different concentration NF-κ B p65 ASOND could restrain HSCs proliferation stimulated by 1 mg/L TNF-α.The expression of I type collagen and mRNA stimulated by 1mg/LTNF-αwas increased,and had a positive correlation with concentration(P>0.05).Conclusion NF-κB p65 ASOND may depress NF-κB activity to restrain HSCs proliferation and Ⅰ type collagen expression,and reduce extracellular matrix.     

Eph受体酪氨酸激酶A2和CD133在胃癌组织的表达及其与胃癌发生发展的关系

目的 探讨Eph受体酪氨酸激酶A2(EphA2)和CD133在胃癌组织中的表达及其与临床病理特征和预后关系.方法 采用免疫组织化学法检测82例胃癌患者手术切除的癌组织和癌旁组织中EphA2和CD133表达水平.结果 EphA2在胃癌组织中的表达率明显高于癌旁组织(86.59％比20.73％,P＜0.01),CD133在胃癌组织中的表达率明显高于癌旁组织(67.07％比10.98％,P＜0.01);EphA2表达水平与胃癌TNM分期、淋巴结转移明显相关(P＜0.05),CD133表达水平与胃癌组织学分化程度、TNM分期、淋巴结转移明显相关(P＜0.05).结论 EphA2和CD133过度表达可能在胃癌的发生、发展过程中发挥作用.     

核因子-κBp65反义寡核苷酸对肝纤维化作用的实验研究

目的 探讨核因子(NF)-κBp65反义寡核苷酸(ASODN)对肝星状细胞(HSC)NF-κB活性和白细胞介素(IL)-6表达的影响.方法 分离培养大鼠HSC,用锥虫蓝染色法和电泳迁移位移试验(EMSA)分别检测NF-κBp65 ASODN对HSC毒性和NF-leB活性的影响,RT-PCR法和ELISA法分别检测对IL-6 mRNA和蛋白表达的影响.结果 ASODN在0.001～1.0μmol/L浓度时,对体外培养的HSC无明显毒性.HSC经肿瘤坏死因子(TNF)-α刺激后,NF-κB活性增强,在0.001～1.0μmol/L的ASODN作用后,NF-κB活性明显减弱.且呈剂量依赖性.同时转染ASODN(0.001～1.0μmol/L)后,TNF-α诱导HSC的IL-6 mRNA及蛋白表达明显降低,亦呈剂量依赖性.结论 ASODN能特异性降低NF-κB活性,同时减少HSC的IL-6表达,为其治疗肝纤维化提供了理论基础.     

Effect of nuclear factor-κB p65 ASOND on I typ collagen expression and rat hepatic stellate cells proliferation

Objective Sstudy effect of nuclear factor-κB ASOND on I type collagen expression and rat hepatic stellate cells(HSC)proliferation.Methods Rat HSCs were separated by affusing and digestingof Ⅳ type collagenenzyme and density acentric method.Lipid-mediated NF-κB p65 ASOND(0.001,0.01,0.1,1μmol/L)Was transferred into rat HSCs.Toxicity of HSCs caused by NF-κB p65 ASOND and activity of LDH were determined by trypan blue staining.Proliferation affection of transferring NF-κB p65 ASOND into HSCs was determined by MTT.In different concentration NF-κB p65 ASOND.expression of Ⅰ type collagen stimulated by 1mg/L TNF-αwas determined by RT-PCR and ELISA.Results After transfection of NF-κB p65 ASODN,expression of NF-κB protein in HSCs was decreasing.Toxicity experiment indicated that NF-κB p65 ASOND of different concentration(0.001,0.01,0.1 and 1.0 μmol/L)had no effect on HSCs livability and LDH activity(P<0.05).Four different concentration NF-κ B p65 ASOND could restrain HSCs proliferation stimulated by 1 mg/L TNF-α.The expression of I type collagen and mRNA stimulated by 1mg/LTNF-αwas increased,and had a positive correlation with concentration(P>0.05).Conclusion NF-κB p65 ASOND may depress NF-κB activity to restrain HSCs proliferation and Ⅰ type collagen expression,and reduce extracellular matrix.     

JS-K 对人结肠癌细胞HCT116 增殖及凋亡的影响

目的 研究JS-K 对人结肠癌细胞HCT116 增殖及凋亡的影响。方法 不同浓度JS-K 处理 HCT116 细胞48 h 后,采用MTS 法检测JS-K 对HCT116 细胞增殖活性的影响;PI 单染流式细胞术检测 JS-K 对HCT116 细胞周期的影响;Annexin V-FITC/PI 双染流式细胞术检测JS-K 对HCT116 细胞凋亡的 影响;Texas Red-X 鬼笔环肽荧光染色观察细胞骨架形态;实时荧光定量聚合酶链反应检测Bcl-2 家族基 因mRNA 表达。结果 JS-K 对人结肠癌细胞HCT116 增殖有抑制作用,且呈浓度和时间依赖性（P <0.05）。 JS-K 诱导HCT116 细胞被阻滞在G2/M 期（P <0.05）。JS-K 可致HCT116 细胞发生晚期凋亡（P <0.05）。 JS-K 可改变HCT116 细胞骨架形态、微丝结构与分布。随着JS-K 浓度增加,促凋亡基因Bax、Bik、Bim、 Puma mRNA 相对表达量上调,抗凋亡基因Mcl-1 mRNA 相对表达量下调（P <0.05）。结论 JS-K 对人结肠 癌HCT116 细胞增殖有明显抑制作用,通过调节Bcl-2 家族基因表达,阻滞细胞G2/M 期,促进细胞发生晚 期凋亡。     

核孔蛋白210与恶性肿瘤关系的研究进展

［摘要］　核孔蛋白210（NUP210）是一种跨膜核孔糖蛋白，参与构成核孔复合物。以往研究认为NUP210只是参与核孔复合物的组装及核膜的分解，但近年来研究发现NUP210与其他生物学功能相关联，且在多种恶性肿瘤中表达上调，与恶性肿瘤的发生、发展密切相关。该文就近年来NUP210与恶性肿瘤关系研究进展作一综述。