Objective： To investigate the possible correlation between osteoglycin expression and gelatinases activity of tumor cells. Methods： Eukaryotic expression plasmid and antisense expression plasmid of osteoglycin were...Objective： To investigate the possible correlation between osteoglycin expression and gelatinases activity of tumor cells. Methods： Eukaryotic expression plasmid and antisense expression plasmid of osteoglycin were constructed, and transfected into mouse hepatocarcinoma Hca-F cells and Hca-P cells, respectively. RT-PCR and Western blot were employed to analyze osteoglycin expression in tumor cells. Zymographic analysis was used to observe the possible correlation between osteoglycin expression and gelatinases activity of tumor cells cultured under different conditions. Results： High expression of osteoglycin after transfection of osteoglycin gene attenuated secretion of gelatinases in Hca-F cells cultured with extract of lymph node （P〈0.05）. However, transfection of osteoglycin gene into Hca-F cells failed to influence gelatinases activity of tumor cells cultured with extracts of liver and spleen or in DMEM medium. Effective suppression of osteoglycin expression in Hca-P cells by osteoglycin shRNA resulted in enhanced activity of gelatinases in Hca-P cells cultured with extract of lymph node （P〈0.05）. However, inhibition of osteoglycin expression in Hca-P cells failed to influence gelatinases activity of tumor cells cultured with extracts of liver and spleen or in DMEM medium. Conclusion： Osteoglycin expression influences gelatinases activity of murine hepatocarcinoma cells cultured with extract of lymph node. Regulation of gelatinases activity might be one of the mechanisms by which osteoglycin contribute to lymphatic metastasis suppression.展开更多
AIM:To investigate the possible correlation between osteoglycin expression and gelatinase activity of mouse hepatocarcinoma Hca-F cells.METHODS:A eukaryotic expression plasmid pIRE Spuro3 osteoglycin(+)was constructed...AIM:To investigate the possible correlation between osteoglycin expression and gelatinase activity of mouse hepatocarcinoma Hca-F cells.METHODS:A eukaryotic expression plasmid pIRE Spuro3 osteoglycin(+)was constructed and transfected into Hca-F cells to investigate the possible correlation between osteoglycin expression and gelatinase activity of Hca-F cells cultured with extract of lymph node,liver,spleen or in DMEM medium.The activity of gelatinases was examined through zymographic analysis.RESULTS:High expression of osteoglycin attenuated the gelatinase activity of Hca-F cells cultured with extract of lymph node,and at the same time,decreased the metastatic potential of Hca-F cells to peripheral lymph nodes in vivo.CONCLUSION:High expression of osteoglycin decreases the gelatinase activity of Hca-F cells cultured with extract of lymph node;regulation of gelatinase activity might be one of mechanisms that osteoglycin contributes to lymphatic metastasis suppression.展开更多
目的探究骨甘蛋白(OGN)对胃癌(GC)细胞干性与化疗耐药性的影响。方法实时荧光聚合酶链反应(RT-PCR)、蛋白质印迹法、免疫组织化学(IHC)联合癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库分析OGN在胃癌细胞和组织中的表达。CCK-8...目的探究骨甘蛋白(OGN)对胃癌(GC)细胞干性与化疗耐药性的影响。方法实时荧光聚合酶链反应(RT-PCR)、蛋白质印迹法、免疫组织化学(IHC)联合癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库分析OGN在胃癌细胞和组织中的表达。CCK-8和克隆形成评估细胞增殖能力变化,通过流式细胞术对细胞凋亡进行测定,CCK-8检测OGN在GC细胞化疗抵抗中的作用。蛋白质印迹法检测细胞干性相关分子标志物表达变化。结果OGN在GC细胞表达低于其在胃黏膜细胞中表达(P<0.05);过表达OGN的GC细胞侵袭能力减弱(P<0.05),凋亡增加(P<0.05)。在铂类和氟尿嘧啶处理后,细胞存活比例下降,化疗抵抗减轻(P<0.05),OGN诱导了化疗耐药。过表达OGN抑制了干性标志物的表达。结论OGN在GC组织中低表达,过表达的OGN能够抑制细胞干性,影响化疗耐药。展开更多
基金supported by the National Natural Science Foundation of China (No.30500586)
文摘Objective： To investigate the possible correlation between osteoglycin expression and gelatinases activity of tumor cells. Methods： Eukaryotic expression plasmid and antisense expression plasmid of osteoglycin were constructed, and transfected into mouse hepatocarcinoma Hca-F cells and Hca-P cells, respectively. RT-PCR and Western blot were employed to analyze osteoglycin expression in tumor cells. Zymographic analysis was used to observe the possible correlation between osteoglycin expression and gelatinases activity of tumor cells cultured under different conditions. Results： High expression of osteoglycin after transfection of osteoglycin gene attenuated secretion of gelatinases in Hca-F cells cultured with extract of lymph node （P〈0.05）. However, transfection of osteoglycin gene into Hca-F cells failed to influence gelatinases activity of tumor cells cultured with extracts of liver and spleen or in DMEM medium. Effective suppression of osteoglycin expression in Hca-P cells by osteoglycin shRNA resulted in enhanced activity of gelatinases in Hca-P cells cultured with extract of lymph node （P〈0.05）. However, inhibition of osteoglycin expression in Hca-P cells failed to influence gelatinases activity of tumor cells cultured with extracts of liver and spleen or in DMEM medium. Conclusion： Osteoglycin expression influences gelatinases activity of murine hepatocarcinoma cells cultured with extract of lymph node. Regulation of gelatinases activity might be one of the mechanisms by which osteoglycin contribute to lymphatic metastasis suppression.
基金Supported by National Natural Science Foundation of China,No.30500586
文摘AIM:To investigate the possible correlation between osteoglycin expression and gelatinase activity of mouse hepatocarcinoma Hca-F cells.METHODS:A eukaryotic expression plasmid pIRE Spuro3 osteoglycin(+)was constructed and transfected into Hca-F cells to investigate the possible correlation between osteoglycin expression and gelatinase activity of Hca-F cells cultured with extract of lymph node,liver,spleen or in DMEM medium.The activity of gelatinases was examined through zymographic analysis.RESULTS:High expression of osteoglycin attenuated the gelatinase activity of Hca-F cells cultured with extract of lymph node,and at the same time,decreased the metastatic potential of Hca-F cells to peripheral lymph nodes in vivo.CONCLUSION:High expression of osteoglycin decreases the gelatinase activity of Hca-F cells cultured with extract of lymph node;regulation of gelatinase activity might be one of mechanisms that osteoglycin contributes to lymphatic metastasis suppression.
文摘目的探究骨甘蛋白(OGN)对胃癌(GC)细胞干性与化疗耐药性的影响。方法实时荧光聚合酶链反应(RT-PCR)、蛋白质印迹法、免疫组织化学(IHC)联合癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库分析OGN在胃癌细胞和组织中的表达。CCK-8和克隆形成评估细胞增殖能力变化,通过流式细胞术对细胞凋亡进行测定,CCK-8检测OGN在GC细胞化疗抵抗中的作用。蛋白质印迹法检测细胞干性相关分子标志物表达变化。结果OGN在GC细胞表达低于其在胃黏膜细胞中表达(P<0.05);过表达OGN的GC细胞侵袭能力减弱(P<0.05),凋亡增加(P<0.05)。在铂类和氟尿嘧啶处理后,细胞存活比例下降,化疗抵抗减轻(P<0.05),OGN诱导了化疗耐药。过表达OGN抑制了干性标志物的表达。结论OGN在GC组织中低表达,过表达的OGN能够抑制细胞干性,影响化疗耐药。
