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国家自然科学基金(30200369)

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相关作者:朱青王立锋张王刚杨安钢王芳更多>>
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Hsp701A的RNA干涉诱导K562细胞凋亡被引量:3
2006年
目的:研究RNA干涉(RNAi)Hsp701A基因的表达及其诱导慢性粒细胞白血病(慢粒)细胞株K562的凋亡。方法:构建Hsp701A基因小干涉RNA(siRNA)的真核表达载体,转染K562细胞并证实RNAi的有效性。用MTT比色法、AnnexinV-FITC/PI染色和流式细胞术,分别检测K562细胞的增殖、凋亡和细胞周期。结果:构建了Hsp701AsiRNA的真核表达载体。将其稳定转染K562细胞后,RNAi组细胞中Hsp701ARNA和蛋白的表达水平明显下降。Hsp701AsiRNA能抑制K562细胞增殖并诱导其细胞凋亡,将其阻滞于G1期。结论:RNAiHsp701A基因诱导的表达有望成为一种新的慢粒的治疗策略。
朱青张王刚王立锋王芳张勇杨安钢
关键词:RNA干涉热休克蛋白70慢性粒细胞白血病细胞凋亡
Gp96多肽复合物激活树突状细胞诱导抗肝癌免疫的研究被引量:2
2004年
目的 研究热休克蛋白Gp96肽复合物修饰树突状细胞(DC)后的体内外特异性抗瘤作用。方法 从小鼠肝癌细胞株H22细胞中提取Gp96肽复合物,采用细胞培养方法,从小鼠骨髓中获得DC,体外大量培养。用Gp96肽复合物修饰DC,刺激小鼠脾淋巴细胞,以51Cr释放法测定细胞毒性T淋巴细胞(CTL)杀伤活性。制作小鼠肝癌模型,分别予以修饰后的DC、Gp96蛋白、灭活的H22肝癌细胞治疗,检测血清中自细胞介素(IL)-10、干扰素(IFN)-γ水平;CD_4、CD_8和IFN-γ、IL-10双阳性细胞比例。观察其抑制肿瘤的效果。结果 Gp96肽复合物修饰的DC可激活小鼠脾淋巴细胞;后者能对H22瘤细胞进行特异性杀伤,而对艾氏腹水瘤细胞无效;经修饰的DC可明显改善机体的免疫状态,使Th1细胞比例上升,抑制体内肿瘤的生长。结论 Gp96肽复合物能够很好的修饰体外诱导获取的DC,使后者成为一种有效的瘤苗。
杨威曹春霞刘青光于良马清涌潘承恩
关键词:热休克蛋白GP96DC树突状细胞鼠肝肝癌
gp96多肽复合物对树突状细胞成熟的影响被引量:1
2005年
目的研究热休克蛋白gp96多肽复合物与小鼠骨髓来源的树突状细胞(DC)成熟的关系。方法采用GM-CSF、IL-4刺激培养小鼠骨髓细胞,增殖产生大量DC;从小鼠肝癌细胞株H22细胞中提取gp96肽复合物,体外修饰DC;通过流式细胞仪检测上清液IL-12、TNF-α细胞因子含量和DC表面主要组织相容性Ⅱ类抗原、CD40、CD80等分子表达;DC与小鼠脾淋巴细胞共同培育后,检测CD4、CD8和IFN-γ、IL-10双阳性细胞比例;以51Cr释放法测定细胞毒性T淋巴细胞(CTL)杀伤活性。结果gp96多肽复合物修饰的DC大量分泌IL-12、TNF-α等细胞因子,高表达MHCⅡ类抗原、CD40、CD80等分子;并且能激活小鼠脾淋巴细胞,CD8+-IFN-γ+和CD4+-IFN-γ+双阳性细胞比例显著升高;能够特异性的杀伤H22癌细胞。结论gp96多肽复合物能够诱导DC成熟,体外产生特异性CTL反应。
杨威曹春霞吴胜利刘青光马清涌潘承恩
关键词:细胞成熟热休克蛋白GP96特异性CTL反应小鼠骨髓细胞流式细胞仪检测
白藜芦醇抗肿瘤活性研究进展被引量:5
2006年
白藜芦醇广泛分布于各种植物及食物中,具有重要的防治肿瘤及心血管疾病的作用,深入研究白藜芦醇的活性,尤其是抗肿瘤活性机制,具有重要的临床意义。
朱青韩苏夏马瑾璐
关键词:白藜芦醇肿瘤心血管疾病
Effect of dendritic cell modified by gp96-peptide complex on antitumor effect in H22 cell
2005年
Objective: To investigate the antitumor effect of dendritic cell (DC) modified by gp96-peptide complexes both in vitro and in vivo. Methods:Gp96-peptide complexes were acquired from H22 liver cancer cells in mice. DC were cultured from bone marrow cells and modified by gp96-peptide complexes. Spleen lymphocytes of mice were activated by modified DC and the cytotoxicity were detected by ^51Cr release method. Modified DC, gp96-peptide complexes and inactivated H22 cells were injected into mice bearing H22 liver cancer cells to observe the levels of IL-10, IFN-y in serum and the alteration of proportions of CD8^+-IFNy^+ and CD8^+-IL-10^+ cells, CD4^+-IFNy^+ and CD4^+-IL-10^+ cells. Results: DC modified by gp96-peptide complexes can activate spleen lymphocyte and the latter can specifically kill H22 cells but not Ehrilich ascites carcinoma cells. Modified DC can improve the host's antitumor immune response and the proportions of Thl cells, inhibiting tumor growth. Conclusion: Gp96-peptide complexes can activate DC effectively, making DC a good vaccine.
石磊岳媛吴胜利张梅潘承恩
Potent antitumor effect elicited by gp96-peptide complexes pulsed by dendritic cell on mice of H22 liver cancer
2006年
Objective: To improve DC-based tumor vaccination, we studied whether dendritic ceils (DCs) which cocultured with H22 liver cancer cells-derived heat shock protein (HSP) glycoprotein 96 (gp96) affect the T cell-activating potential in vitro and the induction of tumor immunity in vivo. Methods: Maturation of murine bone marrow-derived DC was induced by GM-CSF plus IL-4, which mimiced the immunostimulatory effect of DC. Cocultured DC and gp96-peptide complexes were used to vaccine H22 liver cancer cells of mice. Using murine models we compared the immunogenecity of DC modified by gp96-peptides complexes derived from murine liver cancer cells alone or inactive tumor cells. To verify the specificity of the vaccine, in vitro assays were executed. Serum cytokine levels were quantified to explore the supposed pathway of DC modified by gp96 peptide complexes and its effect on antitumor immune response. Results: DC modified by gp96-peptide complexes can activate spleen lymphocyte and the latter can specifically kill H22 cells but not Ehrilich ascites carcinoma cells. Modified DC can induce potent tumor-antigenspecific immune response, augment the proliferation of Thl cells, and inhibit tumor growth. Conclusion: In this study, we have developed a novel DC-mediated tumor vaccine by combing the gp96 antigenic peptides complexes and inducing immune response against specific tumor cells, gp96 can be identified as a potent DC activator.
杨威曹春霞楚雍烈刘青光于良潘承恩
关键词:VACCINE
Hsp701A基因的RNA干涉对HepG2细胞化疗敏感性的影响被引量:3
2006年
目的:研究Hsp701A基因靶向siRNA表达载体对肿瘤细胞化疗药物敏感性的影响。方法:构建Hsp701A靶向小干涉RNA(siRNA)的真核表达载体,并转染HepG2细胞,以证实siRNA真核表达载体RNAi的有效性。用四唑蓝(MTT)比色法检测HepG2细胞对白藜芦醇、顺铂的敏感性。结果:以构建的Hsp701A基因靶向siRNA的真核表达载体稳定转染HepG2细胞后,两种RNAi组细胞中Hsp701ARNA和蛋白的表达水平均明显下降。Hsp701A基因靶向siRNA真核表达载体转染后,HepG2细胞对不同化疗药物的敏感性有不同程度的增加(P<0.05)。结论:Hsp701A基因的RNA干涉可增强HepG2细胞对化疗药物的敏感性。
朱青张王刚王立锋王芳张勇杨安钢
关键词:RNA干涉HEPG2细胞药物敏感性
Heat Shock Protein 96 Induces Maturation of Dendritic Cells被引量:2
2006年
Objective: Heat shock protein (HSP) has the promiscuous abilities to chaperone and present a broad repertoire of tumor antigens to antigen presenting cells including DCs. In this report, we analyzed the modulation of immature DC by lISP 96 (gp96). Method: Murine bone marro6-derived DC was induced by GM-CSF plus IL-4, which aped the immunostimulatory effects of DC. Cocultured DC and gp96-peptide complexes (gp96-PC) or inactivated H22 cells, the expression of MHC class Ⅱ , CD40, CD80 was quantified by flow cytometry. The concentration of IL-12 and TNF- in culture supematants were determined by ELISA. C1 release assay was used to test specific cytotoxic T cell. Results: Our study demonstrated that the extent of DC maturation induced by gp96-PC, which was reflected in surface density of costimulatory and MHC Ⅱmolecules, was correlated with the secretion of IL-12 and with the T cellactivating potential in vitro. Conclusion: Heat shock protein 96 could be isolated and purified from H22 cells and could induce maturation of dendritic cell. Our findings might be relevance to the use of DC vaccine in therapy of human tumors.
Chunxia CaoWei YangYonglie ChuQingguang LiuLiang YuCheng' en Pan
关键词:GP96MATURATION
白藜芦醇诱导肿瘤细胞凋亡和细胞周期阻滞的作用及机制被引量:24
2005年
目的:探讨白藜芦醇(RES)诱导人肝癌HepG2、人慢粒K562肿瘤细胞凋亡和细胞周期阻滞的作用及机制.方法:应用形态学方法,Annexin V荧光染色检测HepG2,K562细胞凋亡的发生,应用流式细胞术(FCM)检测细胞周期,应用四唑蓝比色法(MTT)检测HepG2,K562肿瘤细胞对,RES的药物敏感性.结果:RES对人肝癌HepG2细胞的生长有明显的抑制作用,并诱导肿瘤细胞发生凋亡.凋亡细胞表现为细胞固缩,核染色质碎裂.Annexin V荧光染色检测HepG2细胞凋亡率为33.7%,用FCM检测细胞周期明显阻止于G1期,而K562细胞凋亡率为9.97%.MTT检测表明RES对人肝癌HepG2细胞有剂量-效应关系.但RES对人K562细胞的生长无明显的抑制作用.结论:RES通过诱导HepG2细胞凋亡抑制肿瘤的生长,并有剂量-效应关系.同时RES对肿瘤细胞的抑制及诱导凋亡的作用有细胞选择性.
朱青张王刚王立锋王芳杨安钢
关键词:白藜芦醇细胞凋亡细胞周期HEPG2K562
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