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作 者:左江成[1] 吕建新[1] 金丽琴[2] 邹立林[1] 李东[1] 明镇寰[1]
机构地区:[1]温州医学院细胞与分子医学研究所,浙江温州325035 [2]温州医学院临床医学部生物化学教研室,浙江温州325035
出 处:《中国病理生理杂志》2008年第2期237-241,共5页Chinese Journal of Pathophysiology
基 金:浙江省自然科学基金资助项目(No.301036)
摘 要:目的:通过观察细脚拟青霉多糖的体外抗肿瘤活性,初步探讨其作用机制。方法:采用水提,乙醇沉淀,DEAE-纤维素和Sephadex G-100柱层析分离纯化多糖PTPS-I;以PTPS-I和经PTPS-I刺激培养的人外周血单个核细胞(MNC)的条件培养基(PTPS-I-MNC-CM)作用于体外培养的红白血病细胞K562、喉癌细胞Hep2和肝癌细胞SMMC-7721,观察PTPS-I和PTPS-I-MNC-CM对其增殖活性的影响;用cell counting kit-8(CCK-8)检测MNC的增殖活性;以荧光定量PCR检测MNC中TNF-α和IL-6 mRNA的表达。结果:PTPS-I-MNC-CM能抑制K562,Hep2和SMMC-7721细胞的增殖(均为P<0.01);PTPS-I不能抑制肿瘤细胞的增殖(P>0.05),没有显现直接的细胞毒作用。PTPS-I能促进MNC的增殖(P<0.01)和增强TNF-α和IL-6 mRNA的表达(P<0.05,P<0.01)。结论:PTPS-I具有免疫调节活性,通过免疫调节发挥抗肿瘤作用。AIM: To investigate the antitumor activity and mechanism of Paecilomyces tenuipes polysaccharide (PTPS). METHODS. PTPS-Ⅰ was obtained by water extraction and alcohol precipitation, and purified by DEAE - cellulose and Sephadex G - 100 chromatography. Human erythroleukemia cell line K562, laryngocarcinoma cell line Hep2 and hepatic carcinoma cell line SMMC -7721were co- cultured with PTPS-Ⅰ or the conditioned medium which prepared with PTPS-Ⅰ - stimulated human mononuclear cells ( PTPS-Ⅰ - MNC - CM ), and the proliferation of tumor cells was determined. The cell counting kit - 8 ( CCK - 8) was used to determine the proliferation of MNCs. The FQ - RT - PCR was applied to investigate the expression of TNF - α and IL - 6 mRNA in MNCs. RESULTS: PTPS-Ⅰ - MNC - CM inhibited the proliferation of K562, Hep2 and SMMC -7721 cells in vitro (P 〈0. 01 ). Cytotoxicity of PTPS-Ⅰ against K562, Hep2 and SMMC - 7721 cells was not observed ( P 〈 0.01 ). PTPS-Ⅰ stimulated the proliferation of MNCs ( P 〈 0. 01 ) and significantly enhanced the expression of TNF - α and IL - 6 mRNA in MNCs ( P 〈 0. 05, P 〈 0. 01 ). CONCLUSION : The results suggest that PTPS-Ⅰ is an immunomodulator and its antitumoral activity is through the immunomodulatory mechanism rather than the direct cytotoxicity against tumor cells.
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