AIM: To clarify the role of high in normal-1 (HIN-1) gene promoter methylation during gastric cancer development. METHODS: Gastric cancer cell lines and tissue specimens were analyzed for expression of HIN-1 mRNA and ...AIM: To clarify the role of high in normal-1 (HIN-1) gene promoter methylation during gastric cancer development. METHODS: Gastric cancer cell lines and tissue specimens were analyzed for expression of HIN-1 mRNA and protein using the semi-quantitative reverse transcription polymerase chain reaction and immunohistochemistry. The methylation of the HIN-1 gene promoter was detected in gastric carcinoma cells and tissues using methylation-specific polymerase chain reaction. The 3-(4,5-dimethylthiazol-2yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium cell viability assay and flow cytometry were used to assess the changes in behaviors of gastric cancer cells with or without 5-aza-2’-deoxycytidine treatment. RESULTS: HIN-1 was not expressed in 4 of 5 gastric cancer cell lines. The demethylation reagent 5-aza-2’-deoxycytidine was able to induce or upregulate HIN-1 expression in gastric cancer cell lines, which is associated with reduction of tumor cell viability. Furthermore, methylation of the HIN-1 gene promoter was shown in 57.8% (26/45) of the primary gastric cancer and 42.1% (17/38) of adjacent tissue samples, but was not shown in normal gastric mucosa (0/10). From the clinicopathological data of the patients, methylation of the HIN-1 gene promoter was found to be associated with tumor differentiation (P = 0.000). CONCLUSION: High methylation of HIN-1 gene promoter results in silence of HIN-1 expression in gastric cancer. 5-aza-2’-deoxycytidine reverses HIN-1 methylation and reduces viability of gastric cancer cells.展开更多
目的研究凝血酶对人肺上皮细胞及正常人肺成纤维细胞释放单核细胞趋化蛋白-1(MCP-1)的调节作用。方法人肺支气管上皮细胞(normal human bronchial epithelial cells,NHBE)、人肺癌上皮细胞系A549及正常人肺成纤维细胞(normal human lung...目的研究凝血酶对人肺上皮细胞及正常人肺成纤维细胞释放单核细胞趋化蛋白-1(MCP-1)的调节作用。方法人肺支气管上皮细胞(normal human bronchial epithelial cells,NHBE)、人肺癌上皮细胞系A549及正常人肺成纤维细胞(normal human lung fibroblasts,NH-LF)分别培养于96孔或6孔板培养板内,凝血酶诱导4h和24h后,收集上清液及细胞裂解液。ELISA方法检测MCP-1蛋白水平;实时荧光定量RT-PCR技术检测MCP-1mRNA水平。结果凝血酶可诱导NHBE、A549及NH-LF细胞释放MCP-1,但NH-LF细胞的MCP-1释放量远远高于NHBE及A549细胞;凝血酶还可诱导NH-LF细胞mRNA表达。结论人肺成纤维细胞可能是肺内表达MCP-1的主要来源细胞,因此在肺纤维化时可能起重要作用。展开更多
河套蜜瓜(Cucumis melo L. CV Hetao)是1种典型的呼吸跃变型果实,极其不耐贮藏。本文研究了不同浓度的1-甲基环丙烯(1-MCP)处理对在常温(20℃)贮藏条件下河套蜜瓜品质的影响。结果表明,100nl/l和300nl/l的处理与对照相比由于显著降低呼...河套蜜瓜(Cucumis melo L. CV Hetao)是1种典型的呼吸跃变型果实,极其不耐贮藏。本文研究了不同浓度的1-甲基环丙烯(1-MCP)处理对在常温(20℃)贮藏条件下河套蜜瓜品质的影响。结果表明,100nl/l和300nl/l的处理与对照相比由于显著降低呼吸强度,推迟呼吸跃变时间,从而有效延缓了河套蜜瓜硬度的下降速度,可溶性固形物降解得到抑制,维持了较高的可溶性糖,可滴定酸含量和高的糖酸比,降低了蜜瓜的转黄率、腐烂率及失重率,保持了贮藏期间的品质。500nl/l和1000 nl/l的1-MCP处理因为完全抑制了河套蜜瓜的呼吸使果实不能完熟,从而加重腐烂而影响了贮藏品质。展开更多
基金Supported by National Basic Research Program (973 Program No. 2010CB912802)the Postdoctoral Fund of China, No. 20080441314
文摘AIM: To clarify the role of high in normal-1 (HIN-1) gene promoter methylation during gastric cancer development. METHODS: Gastric cancer cell lines and tissue specimens were analyzed for expression of HIN-1 mRNA and protein using the semi-quantitative reverse transcription polymerase chain reaction and immunohistochemistry. The methylation of the HIN-1 gene promoter was detected in gastric carcinoma cells and tissues using methylation-specific polymerase chain reaction. The 3-(4,5-dimethylthiazol-2yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium cell viability assay and flow cytometry were used to assess the changes in behaviors of gastric cancer cells with or without 5-aza-2’-deoxycytidine treatment. RESULTS: HIN-1 was not expressed in 4 of 5 gastric cancer cell lines. The demethylation reagent 5-aza-2’-deoxycytidine was able to induce or upregulate HIN-1 expression in gastric cancer cell lines, which is associated with reduction of tumor cell viability. Furthermore, methylation of the HIN-1 gene promoter was shown in 57.8% (26/45) of the primary gastric cancer and 42.1% (17/38) of adjacent tissue samples, but was not shown in normal gastric mucosa (0/10). From the clinicopathological data of the patients, methylation of the HIN-1 gene promoter was found to be associated with tumor differentiation (P = 0.000). CONCLUSION: High methylation of HIN-1 gene promoter results in silence of HIN-1 expression in gastric cancer. 5-aza-2’-deoxycytidine reverses HIN-1 methylation and reduces viability of gastric cancer cells.
文摘目的研究凝血酶对人肺上皮细胞及正常人肺成纤维细胞释放单核细胞趋化蛋白-1(MCP-1)的调节作用。方法人肺支气管上皮细胞(normal human bronchial epithelial cells,NHBE)、人肺癌上皮细胞系A549及正常人肺成纤维细胞(normal human lung fibroblasts,NH-LF)分别培养于96孔或6孔板培养板内,凝血酶诱导4h和24h后,收集上清液及细胞裂解液。ELISA方法检测MCP-1蛋白水平;实时荧光定量RT-PCR技术检测MCP-1mRNA水平。结果凝血酶可诱导NHBE、A549及NH-LF细胞释放MCP-1,但NH-LF细胞的MCP-1释放量远远高于NHBE及A549细胞;凝血酶还可诱导NH-LF细胞mRNA表达。结论人肺成纤维细胞可能是肺内表达MCP-1的主要来源细胞,因此在肺纤维化时可能起重要作用。
