AIM:To investigate the promoter methylation status and mRNA expression of DKK-3 and WIF-1 gene in hepatocellular carcinoma(HCC).METHODS:DKK-3 and WIF-1 acted as Wnt-antagonists and tumor suppressors,but hypermethylati...AIM:To investigate the promoter methylation status and mRNA expression of DKK-3 and WIF-1 gene in hepatocellular carcinoma(HCC).METHODS:DKK-3 and WIF-1 acted as Wnt-antagonists and tumor suppressors,but hypermethylation of the gene promoter and low mRNA expression activated Wnt signaling aberrantly and induced the development of HCC.Methylation status of the DKK-3 and WIF-1 gene promoter was investigated using methylation specific polymerase chain reaction(PCR) in tumor and adjacent non-cancerous tissues from 33 HCC patients and 20 normal liver tissues served as control.The expression of DKK-3 and WIF-1 mRNA was also determined by real-time quantitative reverse transcriptase PCR.The relationship between methylation,mRNA expression,and clinical data,as well as methylation and mRNA expression of the two genes were analyzed.RESULTS:The methylation of DKK-3 and WIF-1 genes in HCC increased significantly compared with adjacent non-cancerous tissues and normal control tissues(χ2 =7.79,P < 0.05;χ2 = 4.89,P < 0.05),and no significant difference in methylation between adjacent non-cancerous tissues and normal control tissues was observed.In HCC tissues,significant differences in the DKK-3 promoter methylation were observed in age and cirrhosis,and significant differences of the WIF-1 promoter methylation were observed in HBsAg and cirrhosis.The average expression of DKK-3 mRNA in HCC and adjacent non-cancerous tissues was increased significantly compared with normal control tissues.The average expression of WIF-1 mRNA showed no significant difference among the three tissues.The mRNA expression of DKK-3 gene in HCC was decreased as the pathological grade increased.CONCLUSION:The aberrant promoter methylation and decreased expression of DKK-3 and WIF-1 may be an important mechanism in HCC,and may be a far-reaching significance in early diagnosis and therapy of HCC.展开更多
目曲：检测dickkopf（DKK1）在肝癌组织及多种恶性肿瘤细胞系中的表达情况。方法：采用酶联免疫吸附实验、real time PCR、RT—PCR及Western blot对DKK1在肝癌组织及多种恶性肿瘤细胞系的表达情况进行检测。结果：肝癌组织检测RT—PCR...目曲：检测dickkopf（DKK1）在肝癌组织及多种恶性肿瘤细胞系中的表达情况。方法：采用酶联免疫吸附实验、real time PCR、RT—PCR及Western blot对DKK1在肝癌组织及多种恶性肿瘤细胞系的表达情况进行检测。结果：肝癌组织检测RT—PCR，10／15肝癌中高表达；Real—time PCR方法检测，8／8肝癌中高表达；Western blot方法检测，5／8肝癌中高表达。酶联免疫吸附实验检测到DKK1在多种恶性肿瘤细胞系的细胞培养液上清液中均有表达，蛋白质表达量波动于5～210ng／mL之间；10种人恶性肿瘤细胞系在mRNA水平和蛋白质水平DKK1的表达均能检测到。结论：DKK1高表达于肝癌组织及广泛表达于多种恶性肿瘤细胞系，提示DKK1可能在肿瘤的发生、发展中起到重要作用。展开更多
背景:中医药可有效防治糖皮质激素性骨质疏松,但其防治机制尚不明确。DKK1是Wnt/β-catenin信号通路的抑制剂,糖皮质激素可诱导其上调。因此,DKK1蛋白是防治糖皮质激素性骨质疏松的重要靶点。目的:探讨左归丸防治糖皮质激素性骨质疏松中...背景:中医药可有效防治糖皮质激素性骨质疏松,但其防治机制尚不明确。DKK1是Wnt/β-catenin信号通路的抑制剂,糖皮质激素可诱导其上调。因此,DKK1蛋白是防治糖皮质激素性骨质疏松的重要靶点。目的:探讨左归丸防治糖皮质激素性骨质疏松中对DKK1的调控作用。方法:18只3月龄雌性SD大鼠随机均分为3组:空白组、模型组和左归丸组。模型组和左归丸组大鼠皮下注射地塞米松建立糖皮质激素性骨质疏松模型;空白组给予等体积生理盐水;左归丸组皮下注射地塞米松同时给予左归丸水提液灌胃。1个月后取大鼠腰椎进行micro-CT检测骨量及骨微细结构、压缩试验检测生物力学性能,q PCR测定腰椎DKK1、Runx2和CTSK m RNA表达;血清检测碱性磷酸酶活性。结果与结论:(1)与空白组比较,模型组体积骨密度、相对骨体积、骨小梁数量和骨小梁厚度显著降低(P<0.05),骨小梁间距和结构模型指数较空白组显著增大(P<0.05),血清碱性磷酸酶活性较空白组呈降低趋势,DKK1 m RNA表达显著上调(P<0.05),成骨相关因子Runx2 m RNA表达呈下调趋势,破骨相关因子CTSK m RNA呈上调趋势;(2)与模型组比较,左归丸组体积骨密度、相对骨体积和骨小梁数量显著提升(P<0.05),结构模型指数显著降低(P<0.05),骨小梁间距有减小趋势但差异无显著性意义,血清碱性磷酸酶活性较模型组呈升高趋势,DKK1 m RNA表达显著下调(P<0.05),Runx2 m RNA表达呈上调趋势,CTSK m RNA呈下调趋势;(3)与模型组比较,左归丸组椎体压缩强度显著提升(P<0.05);(4)结果说明,左归丸可能通过下调DKK1的表达防治糖皮质激素性骨质疏松。展开更多
基金Supported by The Natural Science Fund of Educational Department of Anhui Province,No. 2006KJ094A
文摘AIM:To investigate the promoter methylation status and mRNA expression of DKK-3 and WIF-1 gene in hepatocellular carcinoma(HCC).METHODS:DKK-3 and WIF-1 acted as Wnt-antagonists and tumor suppressors,but hypermethylation of the gene promoter and low mRNA expression activated Wnt signaling aberrantly and induced the development of HCC.Methylation status of the DKK-3 and WIF-1 gene promoter was investigated using methylation specific polymerase chain reaction(PCR) in tumor and adjacent non-cancerous tissues from 33 HCC patients and 20 normal liver tissues served as control.The expression of DKK-3 and WIF-1 mRNA was also determined by real-time quantitative reverse transcriptase PCR.The relationship between methylation,mRNA expression,and clinical data,as well as methylation and mRNA expression of the two genes were analyzed.RESULTS:The methylation of DKK-3 and WIF-1 genes in HCC increased significantly compared with adjacent non-cancerous tissues and normal control tissues(χ2 =7.79,P < 0.05;χ2 = 4.89,P < 0.05),and no significant difference in methylation between adjacent non-cancerous tissues and normal control tissues was observed.In HCC tissues,significant differences in the DKK-3 promoter methylation were observed in age and cirrhosis,and significant differences of the WIF-1 promoter methylation were observed in HBsAg and cirrhosis.The average expression of DKK-3 mRNA in HCC and adjacent non-cancerous tissues was increased significantly compared with normal control tissues.The average expression of WIF-1 mRNA showed no significant difference among the three tissues.The mRNA expression of DKK-3 gene in HCC was decreased as the pathological grade increased.CONCLUSION:The aberrant promoter methylation and decreased expression of DKK-3 and WIF-1 may be an important mechanism in HCC,and may be a far-reaching significance in early diagnosis and therapy of HCC.
文摘背景:中医药可有效防治糖皮质激素性骨质疏松,但其防治机制尚不明确。DKK1是Wnt/β-catenin信号通路的抑制剂,糖皮质激素可诱导其上调。因此,DKK1蛋白是防治糖皮质激素性骨质疏松的重要靶点。目的:探讨左归丸防治糖皮质激素性骨质疏松中对DKK1的调控作用。方法:18只3月龄雌性SD大鼠随机均分为3组:空白组、模型组和左归丸组。模型组和左归丸组大鼠皮下注射地塞米松建立糖皮质激素性骨质疏松模型;空白组给予等体积生理盐水;左归丸组皮下注射地塞米松同时给予左归丸水提液灌胃。1个月后取大鼠腰椎进行micro-CT检测骨量及骨微细结构、压缩试验检测生物力学性能,q PCR测定腰椎DKK1、Runx2和CTSK m RNA表达;血清检测碱性磷酸酶活性。结果与结论:(1)与空白组比较,模型组体积骨密度、相对骨体积、骨小梁数量和骨小梁厚度显著降低(P<0.05),骨小梁间距和结构模型指数较空白组显著增大(P<0.05),血清碱性磷酸酶活性较空白组呈降低趋势,DKK1 m RNA表达显著上调(P<0.05),成骨相关因子Runx2 m RNA表达呈下调趋势,破骨相关因子CTSK m RNA呈上调趋势;(2)与模型组比较,左归丸组体积骨密度、相对骨体积和骨小梁数量显著提升(P<0.05),结构模型指数显著降低(P<0.05),骨小梁间距有减小趋势但差异无显著性意义,血清碱性磷酸酶活性较模型组呈升高趋势,DKK1 m RNA表达显著下调(P<0.05),Runx2 m RNA表达呈上调趋势,CTSK m RNA呈下调趋势;(3)与模型组比较,左归丸组椎体压缩强度显著提升(P<0.05);(4)结果说明,左归丸可能通过下调DKK1的表达防治糖皮质激素性骨质疏松。
