目的:探讨在髓母细胞瘤(medulloblastoma,MB)中磷酸化修饰对Suppressor of fused(Sufu)抑癌活性的调节和潜在机制。方法:免疫组化分析MB组织芯片中Sufu的表达及磷酸化水平。在MB细胞系DAOY中转染野生型Sufu或其磷酸化位点突变质粒,分别...目的:探讨在髓母细胞瘤(medulloblastoma,MB)中磷酸化修饰对Suppressor of fused(Sufu)抑癌活性的调节和潜在机制。方法:免疫组化分析MB组织芯片中Sufu的表达及磷酸化水平。在MB细胞系DAOY中转染野生型Sufu或其磷酸化位点突变质粒,分别通过细胞免疫化学染色、Western blot、CCK8、EdU标记、细胞流式检测等分析Sufu的核浆分布以及转染细胞的增殖、凋亡等指标,通过荧光素酶报告基因实验检测转染细胞Hedgehog(Hh)信号通路下游胶质瘤相关癌基因同源物(hlioma-associated oncogene homologue,Gli)的转录活性,Western blot检测鸟氨酸脱羧酶(ornithine decarboxylase,ODC1)的表达水平。结果:Sufu在MB组织的表达明显高于正常小脑,并呈现磷酸化的修饰状态。磷酸化Sufu主要聚集在细胞核。Sufu能抑制DAOY细胞的增殖及克隆形成,促进细胞凋亡。非磷酸化的Sufu突变体抑癌作用明显减弱。已知在Hh通路下游,Gli介导的转录活化以及ODC1介导的多胺合成增加均可以促进MB细胞的生长。磷酸化Sufu抑制Gli介导的转录活化,非磷酸化Sufu对Gli的抑制作用减弱,并促进ODC1的表达。结论:Sufu发生磷酸化修饰后,对MB的抑制作用增强,其机制可能与抑制Hh信号下游的Gli转导活性和细胞的多胺代谢有关。展开更多
Traditional Chinese red sufu is a popular condiment,and typical flavor is an important indicator of sensory qualities in commercial products.In this study,the volatile aroma analysis of Wangzhihe red sufu(WRS)from dif...Traditional Chinese red sufu is a popular condiment,and typical flavor is an important indicator of sensory qualities in commercial products.In this study,the volatile aroma analysis of Wangzhihe red sufu(WRS)from different fermentation stages and four typical red sufu products from different locations was carried out by gas chromatography-mass spectrometry(GC–MS),gas chromatography-mass spectrometry/olfactory(GC–MS/O),electronic nose(E-nose)and sensory evaluation.Results showed that 106 volatile compounds were identified in Wangzhihe red sufu during fermentation process,in which phenolics and alcohols were dominant at molded and salted phetze stages,while esters and alcohols became predominate at post-fermentation stage.The volatile aroma substances varied at each fermentation stage.Furthermore,86 volatiles,including 16 aroma-active compounds,were detected in four typical red sufu products.The multivariate analysis results showed the difference in samples from different fermentation stages and typical red sufu products according to GC–MS and E-nose analysis.Combined with flavor omics,discriminant model was established for effective discrimination of samples from different fermentation stages and locations,as well as references to sufu maturity extent.The study presented a new strategy for quality evaluation of red sufu,which could be the supplement to quality evaluation standards.展开更多
Samples of sufu, a traditional food product made by fermenting soybeans, were collected in the city of Anshun in Southwest China and their bacterial diversity was investigated. Nine samples were divided into groups A ...Samples of sufu, a traditional food product made by fermenting soybeans, were collected in the city of Anshun in Southwest China and their bacterial diversity was investigated. Nine samples were divided into groups A C, according to the markets in which they were collected, and the samples were subjected to pyrosequencing of the 16S rRNA V3 V4 regions for evaluation of the bacterial community. A total of 342 operational taxonomic units (OTUs) were determined. One of the samples (B2) included as many as 286 OTUs, and it also displayed the greatest bacterial richness. The most abundant bacterial diversity was shown in group B. F irmicutes was the dominant phylum in groups A and B, while P roteobacteria was the dominant phylum in group C. L actobacillus was the prevailing genus in group A, T etragenococcus in group B, and E mpedobacter in group C. The bacterial composition of sufu depends, to a large degree, on the manufacturer. The obtained results showed regional di erences in the bacterial community patterns of sufu products, which could contribute to a better understanding of the fermentation and ripening bioprocesses of sufu.展开更多
Objective： to investigate the expressions of beta-catenin, SUFU and VEGFR-2 proteins in medulloblastoma. Methods： Immunohistochemical staining with SP method was conducted to determine the expressions of beta-cateni...Objective： to investigate the expressions of beta-catenin, SUFU and VEGFR-2 proteins in medulloblastoma. Methods： Immunohistochemical staining with SP method was conducted to determine the expressions of beta-catenin, SUFU and VEGFR-2 in 33 cases of medulloblastoma and 10 normal cerebellar tissues. Results： the abnormal expression rates of beta-catenin and VEGFR-2 in medulloblastoma were significantly higher than that in normal tissue. While the positive expression of SUFU gene in medulloblastoma was significantly lower than that in 10 normal cerebellar tissues, A significant negative correlation was found between beta-catenin and SUFU proteins and a positive correlation between beta-catenin and VEGFR-2 was found in medulloblastoma. Conclusion： Beta-catenin, VEGFR-2 and SUFU have important effects on the pathogenesis and development of medulloblastoma.展开更多
Hedgehog信号通路在胚胎发育、组织再生中发挥重要的作用,且与癌症发生发展密切相关.其胞内调控组分Suppressor of Fused(SuFu)蛋白通过结合转录因子Gli(s),负调控该信号通路,但其作用的分子机制仍不甚清楚.在本项研究中,以人SuFu作为...Hedgehog信号通路在胚胎发育、组织再生中发挥重要的作用,且与癌症发生发展密切相关.其胞内调控组分Suppressor of Fused(SuFu)蛋白通过结合转录因子Gli(s),负调控该信号通路,但其作用的分子机制仍不甚清楚.在本项研究中,以人SuFu作为诱饵蛋白,利用酵母双杂交技术成功地筛选到1个新的相互作用因子—蛋白酶体成熟蛋白(POMP).通过免疫共沉淀、体外GST pull-down和免疫细胞化学实验验证其相互作用.为了探究POMP与SuFu的相互作用对Hedgehog信号通路的影响,构建了POMP的过表达质粒和干扰质粒(miR-RNAi)以及转录因子Gli活性检测系统,即荧光素酶报告基因法,结果显示,过表达SuFu蛋白时POMP正调控Hedgehog信号通路,而下调POMP的表达则抑制Gli的活性.该研究结果揭示了POMP新的生物学功能,为阐明Hedgehog信号通路的具体分子机制提供了新的线索.展开更多
以Hedgehog信号通路关键负调控因子Suppressor of Fused(SuFu)蛋白作为诱饵蛋白,利用酵母双杂交系统从人均一化cDNA文库中筛选与之相互作用的蛋白质,共获得160个阳性克隆,包括Hedgehog信号通路蛋白Gli3和一些肿瘤蛋白如乙醛脱氢酶1(ALDH...以Hedgehog信号通路关键负调控因子Suppressor of Fused(SuFu)蛋白作为诱饵蛋白,利用酵母双杂交系统从人均一化cDNA文库中筛选与之相互作用的蛋白质,共获得160个阳性克隆,包括Hedgehog信号通路蛋白Gli3和一些肿瘤蛋白如乙醛脱氢酶1(ALDH1A1)、激酶、转运蛋白、转录因子等.通过免疫共沉淀和免疫细胞化学实验证实,SuFu与ALDH1A1发生相互作用,后者可能参与Hedgehog信号通路的调控.展开更多
基金the National Key R&D program of China(2016YFD0400500)Beijing Postdoctoral Research Foundation(2018-ZZ-120)Chinese Academy of Engineering Project(2018-XY-28).
文摘Traditional Chinese red sufu is a popular condiment,and typical flavor is an important indicator of sensory qualities in commercial products.In this study,the volatile aroma analysis of Wangzhihe red sufu(WRS)from different fermentation stages and four typical red sufu products from different locations was carried out by gas chromatography-mass spectrometry(GC–MS),gas chromatography-mass spectrometry/olfactory(GC–MS/O),electronic nose(E-nose)and sensory evaluation.Results showed that 106 volatile compounds were identified in Wangzhihe red sufu during fermentation process,in which phenolics and alcohols were dominant at molded and salted phetze stages,while esters and alcohols became predominate at post-fermentation stage.The volatile aroma substances varied at each fermentation stage.Furthermore,86 volatiles,including 16 aroma-active compounds,were detected in four typical red sufu products.The multivariate analysis results showed the difference in samples from different fermentation stages and typical red sufu products according to GC–MS and E-nose analysis.Combined with flavor omics,discriminant model was established for effective discrimination of samples from different fermentation stages and locations,as well as references to sufu maturity extent.The study presented a new strategy for quality evaluation of red sufu,which could be the supplement to quality evaluation standards.
文摘Samples of sufu, a traditional food product made by fermenting soybeans, were collected in the city of Anshun in Southwest China and their bacterial diversity was investigated. Nine samples were divided into groups A C, according to the markets in which they were collected, and the samples were subjected to pyrosequencing of the 16S rRNA V3 V4 regions for evaluation of the bacterial community. A total of 342 operational taxonomic units (OTUs) were determined. One of the samples (B2) included as many as 286 OTUs, and it also displayed the greatest bacterial richness. The most abundant bacterial diversity was shown in group B. F irmicutes was the dominant phylum in groups A and B, while P roteobacteria was the dominant phylum in group C. L actobacillus was the prevailing genus in group A, T etragenococcus in group B, and E mpedobacter in group C. The bacterial composition of sufu depends, to a large degree, on the manufacturer. The obtained results showed regional di erences in the bacterial community patterns of sufu products, which could contribute to a better understanding of the fermentation and ripening bioprocesses of sufu.
文摘Objective： to investigate the expressions of beta-catenin, SUFU and VEGFR-2 proteins in medulloblastoma. Methods： Immunohistochemical staining with SP method was conducted to determine the expressions of beta-catenin, SUFU and VEGFR-2 in 33 cases of medulloblastoma and 10 normal cerebellar tissues. Results： the abnormal expression rates of beta-catenin and VEGFR-2 in medulloblastoma were significantly higher than that in normal tissue. While the positive expression of SUFU gene in medulloblastoma was significantly lower than that in 10 normal cerebellar tissues, A significant negative correlation was found between beta-catenin and SUFU proteins and a positive correlation between beta-catenin and VEGFR-2 was found in medulloblastoma. Conclusion： Beta-catenin, VEGFR-2 and SUFU have important effects on the pathogenesis and development of medulloblastoma.
文摘Hedgehog信号通路在胚胎发育、组织再生中发挥重要的作用,且与癌症发生发展密切相关.其胞内调控组分Suppressor of Fused(SuFu)蛋白通过结合转录因子Gli(s),负调控该信号通路,但其作用的分子机制仍不甚清楚.在本项研究中,以人SuFu作为诱饵蛋白,利用酵母双杂交技术成功地筛选到1个新的相互作用因子—蛋白酶体成熟蛋白(POMP).通过免疫共沉淀、体外GST pull-down和免疫细胞化学实验验证其相互作用.为了探究POMP与SuFu的相互作用对Hedgehog信号通路的影响,构建了POMP的过表达质粒和干扰质粒(miR-RNAi)以及转录因子Gli活性检测系统,即荧光素酶报告基因法,结果显示,过表达SuFu蛋白时POMP正调控Hedgehog信号通路,而下调POMP的表达则抑制Gli的活性.该研究结果揭示了POMP新的生物学功能,为阐明Hedgehog信号通路的具体分子机制提供了新的线索.
文摘以Hedgehog信号通路关键负调控因子Suppressor of Fused(SuFu)蛋白作为诱饵蛋白,利用酵母双杂交系统从人均一化cDNA文库中筛选与之相互作用的蛋白质,共获得160个阳性克隆,包括Hedgehog信号通路蛋白Gli3和一些肿瘤蛋白如乙醛脱氢酶1(ALDH1A1)、激酶、转运蛋白、转录因子等.通过免疫共沉淀和免疫细胞化学实验证实,SuFu与ALDH1A1发生相互作用,后者可能参与Hedgehog信号通路的调控.
