Auxin plays critical roles in root formation and development. The components involved in this process, however, are not well understood. Here, we newly identified a peptide encoding gene, auxin-responsive endogenous p...Auxin plays critical roles in root formation and development. The components involved in this process, however, are not well understood. Here, we newly identified a peptide encoding gene, auxin-responsive endogenous polypeptide 1 （AREP1）, which is induced by auxin, and mediates root development in Arabidopsis. Expression of AREP1 was specific to the cotyledon and to root and shoot meristem tissues. Amounts of AREP1 transcripts and AREP1-green fluorescent protein fusion proteins were elevated in response to indoleacetic acid treatment. Suppression of AREP1 through RNAi silencing resulted in reduction of primary root length, increase of lateral root number, and expansion of adventitious roots, compared to the observations in wild-type plants in the presence of auxin. By contrast, transgenic plants overexpressing AREP1 showed enhanced growth of the primary root under auxin treatment. Additionally, rootmorphology, including lateral root number and adventitious roots, differed greatly between transgenic and wildtype plants. Further analysis indicated that the expression of auxin-responsive genes, such as IAA3, IAA7, IAA17, GH3.2, GH3.3, and SAUR-AC1, was significantly higher in AREP1 RNAi plants, and was slightly lower in AREP1 overexpressing plants than in wildtype plants. These results suggest that the novel endogenous peptide AREP1 plays an important role in the process of auxinmediated root development.展开更多
目的通过多肽组学分析,从子宫内膜癌组织中筛选获得具有潜在生物学活性的内源性多肽。方法收集3例子宫内膜样腺癌组织标本(以3例年龄匹配的健康女性子宫内膜样本为对照),通过液相色谱-串联质谱法(liquid chromatography-tandem mass spe...目的通过多肽组学分析,从子宫内膜癌组织中筛选获得具有潜在生物学活性的内源性多肽。方法收集3例子宫内膜样腺癌组织标本(以3例年龄匹配的健康女性子宫内膜样本为对照),通过液相色谱-串联质谱法(liquid chromatography-tandem mass spectrometry,LC-MS/MS)获得子宫内膜癌特异性内源性多肽,借助生物信息学方法筛选具有潜在活性的功能多肽,使用CCK-8实验、Transwell及划痕实验验证多肽对子宫内膜癌细胞株HEC-1-A增殖、侵袭、迁移的影响,并以PCR、Western Blot方法选择性观察多肽对Wnt/β-catenin/EMT通路相关分子表达的影响。结果鉴定获得序列为TAGNVLMH的多肽(PDAEP-1)对应前体为SLC38A10,其表达水平与子宫内膜癌患者5年生存率相关;该多肽对子宫内膜癌细胞的增殖、侵袭、迁移有明显的抑制作用,且作用后Vimentin、β-catenin两者的mRNA及蛋白水平均显著降低,而E-cadherin则上调。结论通过调变Wnt/β-catenin/EMT通路相关分子,内源性多肽PDAEP-1在体外有抑制子宫内膜癌细胞增殖、侵袭和迁移的抗肿瘤活性,具有潜在的诊疗应用价值。展开更多
目的探讨内源性多肽ECDP4在人外周血单核细胞(PBMCs)和子宫内膜癌Ishikawa细胞共培养体系下对PBMCs中T淋巴细胞亚群比例的影响。方法收集2020年9月至2020年11月手术切除的3例子宫内膜癌和3例配对正常子宫内膜组织标本。使用在线数据库Un...目的探讨内源性多肽ECDP4在人外周血单核细胞(PBMCs)和子宫内膜癌Ishikawa细胞共培养体系下对PBMCs中T淋巴细胞亚群比例的影响。方法收集2020年9月至2020年11月手术切除的3例子宫内膜癌和3例配对正常子宫内膜组织标本。使用在线数据库UniProt、ProteomicsDB、MaxQB、UniProt和The Human Protein Atlas分析多肽ECDP4的前体蛋白及其生物学功能。建立人子宫内膜癌Ishikawa细胞株和新鲜人PBMCs共培养模型,分为对照组(含5%胎牛血清培养基)和实验组(含5%胎牛血清培养基中加100 ng/L内源性多肽ECDP4),共培养96 h,收集PBMCs。用流式细胞术分析PBMCs中淋巴细胞亚群比例变化。结果在正常子宫内膜组织中,ECDP4表达量为7.33×10-4±1.13×10-4,而在子宫内膜癌组织中表达量为7.30×10^(-5)±8.47×10^(-6)(P=0.0016)。生物信息学分析显示,NOVA2是ECDP4的前体蛋白,但其并不是子宫内膜癌的预后因素。流式细胞术检测显示,相较于单纯培养PBMCs,共培养体系中CD4^(+)T淋巴细胞比例下调(51.31±11.05)%(P<0.001),CD8^(+)T淋巴细胞比例下调(52.98±5.86)%(P<0.001)。相较于对照组,在共培养体系中加入ECDP4后,CD4^(+)T淋巴细胞比例上调(15.58±2.34)%(P<0.001),Ki-67的比例上调(54.60±24.5)%(P=0.0043),干扰素γ(INF-γ)的比例上调(27.92±17.25)%(P=0.0486);CD8^(+)T淋巴细胞比例上调(20.04±6.70)%(P<0.001),Ki-67的比例上调(26.57±6.42)%(P<0.001),INF-γ比例上调(87.48±40.70)%(P=0.01)。结论多肽ECDP4作为NOVA2的降解产物在子宫内膜癌中显著低表达,其能够增加PBMCs中CD4^(+)T淋巴细胞及CD8^(+)T淋巴细胞的比例,促进CD4^(+)及CD8^(+)T淋巴细胞的增殖作用,同时增加INF-γ的分泌量。内源性多肽ECDP4具有进一步应用于临床研究的价值。展开更多
基金funded by the National Natural Science Foundation of China(30971557,30971816,and31300996)the Guangdong Natural Science Foundation(S2011010001433)
文摘Auxin plays critical roles in root formation and development. The components involved in this process, however, are not well understood. Here, we newly identified a peptide encoding gene, auxin-responsive endogenous polypeptide 1 （AREP1）, which is induced by auxin, and mediates root development in Arabidopsis. Expression of AREP1 was specific to the cotyledon and to root and shoot meristem tissues. Amounts of AREP1 transcripts and AREP1-green fluorescent protein fusion proteins were elevated in response to indoleacetic acid treatment. Suppression of AREP1 through RNAi silencing resulted in reduction of primary root length, increase of lateral root number, and expansion of adventitious roots, compared to the observations in wild-type plants in the presence of auxin. By contrast, transgenic plants overexpressing AREP1 showed enhanced growth of the primary root under auxin treatment. Additionally, rootmorphology, including lateral root number and adventitious roots, differed greatly between transgenic and wildtype plants. Further analysis indicated that the expression of auxin-responsive genes, such as IAA3, IAA7, IAA17, GH3.2, GH3.3, and SAUR-AC1, was significantly higher in AREP1 RNAi plants, and was slightly lower in AREP1 overexpressing plants than in wildtype plants. These results suggest that the novel endogenous peptide AREP1 plays an important role in the process of auxinmediated root development.
文摘目的通过多肽组学分析,从子宫内膜癌组织中筛选获得具有潜在生物学活性的内源性多肽。方法收集3例子宫内膜样腺癌组织标本(以3例年龄匹配的健康女性子宫内膜样本为对照),通过液相色谱-串联质谱法(liquid chromatography-tandem mass spectrometry,LC-MS/MS)获得子宫内膜癌特异性内源性多肽,借助生物信息学方法筛选具有潜在活性的功能多肽,使用CCK-8实验、Transwell及划痕实验验证多肽对子宫内膜癌细胞株HEC-1-A增殖、侵袭、迁移的影响,并以PCR、Western Blot方法选择性观察多肽对Wnt/β-catenin/EMT通路相关分子表达的影响。结果鉴定获得序列为TAGNVLMH的多肽(PDAEP-1)对应前体为SLC38A10,其表达水平与子宫内膜癌患者5年生存率相关;该多肽对子宫内膜癌细胞的增殖、侵袭、迁移有明显的抑制作用,且作用后Vimentin、β-catenin两者的mRNA及蛋白水平均显著降低,而E-cadherin则上调。结论通过调变Wnt/β-catenin/EMT通路相关分子,内源性多肽PDAEP-1在体外有抑制子宫内膜癌细胞增殖、侵袭和迁移的抗肿瘤活性,具有潜在的诊疗应用价值。
文摘目的探讨内源性多肽ECDP4在人外周血单核细胞(PBMCs)和子宫内膜癌Ishikawa细胞共培养体系下对PBMCs中T淋巴细胞亚群比例的影响。方法收集2020年9月至2020年11月手术切除的3例子宫内膜癌和3例配对正常子宫内膜组织标本。使用在线数据库UniProt、ProteomicsDB、MaxQB、UniProt和The Human Protein Atlas分析多肽ECDP4的前体蛋白及其生物学功能。建立人子宫内膜癌Ishikawa细胞株和新鲜人PBMCs共培养模型,分为对照组(含5%胎牛血清培养基)和实验组(含5%胎牛血清培养基中加100 ng/L内源性多肽ECDP4),共培养96 h,收集PBMCs。用流式细胞术分析PBMCs中淋巴细胞亚群比例变化。结果在正常子宫内膜组织中,ECDP4表达量为7.33×10-4±1.13×10-4,而在子宫内膜癌组织中表达量为7.30×10^(-5)±8.47×10^(-6)(P=0.0016)。生物信息学分析显示,NOVA2是ECDP4的前体蛋白,但其并不是子宫内膜癌的预后因素。流式细胞术检测显示,相较于单纯培养PBMCs,共培养体系中CD4^(+)T淋巴细胞比例下调(51.31±11.05)%(P<0.001),CD8^(+)T淋巴细胞比例下调(52.98±5.86)%(P<0.001)。相较于对照组,在共培养体系中加入ECDP4后,CD4^(+)T淋巴细胞比例上调(15.58±2.34)%(P<0.001),Ki-67的比例上调(54.60±24.5)%(P=0.0043),干扰素γ(INF-γ)的比例上调(27.92±17.25)%(P=0.0486);CD8^(+)T淋巴细胞比例上调(20.04±6.70)%(P<0.001),Ki-67的比例上调(26.57±6.42)%(P<0.001),INF-γ比例上调(87.48±40.70)%(P=0.01)。结论多肽ECDP4作为NOVA2的降解产物在子宫内膜癌中显著低表达,其能够增加PBMCs中CD4^(+)T淋巴细胞及CD8^(+)T淋巴细胞的比例,促进CD4^(+)及CD8^(+)T淋巴细胞的增殖作用,同时增加INF-γ的分泌量。内源性多肽ECDP4具有进一步应用于临床研究的价值。
