[Objective]The study aimed to screen the starch-degrading bacterium in bagasse and carry on the identification of strains s2g5-1 and s3g4-8.[Method]By using a variety of selective media,varieties of starch degrading b...[Objective]The study aimed to screen the starch-degrading bacterium in bagasse and carry on the identification of strains s2g5-1 and s3g4-8.[Method]By using a variety of selective media,varieties of starch degrading bacterium were isolated from the sugar cane bagasse form different stages of natural fermentation,then,primary screening and secondary screening were performed.[Result] Starch-degrading strains s2g5-1 and s3g4-8 were screened,and they were identified as Bacillus amyloliquefaciens according to their morphological,physiological,biochemical and molecular characteristics.[Conclusion]The research provided theoretical basis for factory application of bagasse.展开更多
目的:验证G2/S期应答相关蛋白1(G2 and S phase-expressed protein 1,GTSE1)在原发性肝癌组织及细胞株中高表达,并探讨其表达下调对肝癌细胞凋亡的影响。方法:采用内置R程序对GEO(Gene Expression Omnibus)数据库中4项原发性肝癌临床研...目的:验证G2/S期应答相关蛋白1(G2 and S phase-expressed protein 1,GTSE1)在原发性肝癌组织及细胞株中高表达,并探讨其表达下调对肝癌细胞凋亡的影响。方法:采用内置R程序对GEO(Gene Expression Omnibus)数据库中4项原发性肝癌临床研究的基因表达矩阵数据进行分析,筛选肝癌与癌旁组织差异表达基因。采用蛋白质印迹法验证3例肝癌组织和3株肝癌细胞中GTSE1蛋白表达水平。向肝癌Hep-G2和Bel-7402细胞中分别转染GTSE1 siRNA后,采用CCK-8和FCM法分别检测GTSE1表达下调对肝癌细胞增殖、凋亡及5-氟尿嘧啶(5-fluorouracil,5-FU)敏感性的影响,进一步采用蛋白质印迹法检测p53含量及其亚细胞定位,以及Bcl-2家族成员的表达变化。结果:4项研究中原发性肝癌组织表达水平均上调和下调的基因分别为51和146个。其中,上调基因产物GTSE1蛋白在3例临床肝癌组织标本和3株肝癌细胞Hep-G2、Bel-7402和SMMC-7721中均得到进一步验证(P值均<0.001)。siRNA转染后,肝癌Hep-G2和Bel-7402细胞中GTSE1基因沉默效率>70%。与对照组相比,GTSE1表达下调后肝癌Hep-G2和Bel-7402细胞增殖被明显抑制,并随着时间延长,抑制作用逐渐增强(P值均<0.05);GTSE1表达下调后肝癌Hep-G2和Bel-7402细胞周期被阻滞在G1期,同时细胞凋亡率明显升高,且细胞对5-FU的敏感性明显增强(P值均<0.01)。GTSE1表达下调后的肝癌Hep-G2和Bel-7402细胞中总p53及磷酸化P53表达水平均明显上调(P值均<0.05),且细胞核内p53水平升高(P值均<0.01);Bcl-2家族促凋亡成员Bax和Bak表达水平均上调,而抗凋亡成员Bcl-2表达水平下调(P值均<0.05)。结论:下调肝癌中GTSE1蛋白表达能通过活化p53通路促进细胞凋亡,并增加肝癌细胞对化疗药物的敏感性。展开更多
基金Supported by the Central Public-interest Scientific Institution Basal Research Fund(2008hzs1J021,2009hzs1J033)~~
文摘[Objective]The study aimed to screen the starch-degrading bacterium in bagasse and carry on the identification of strains s2g5-1 and s3g4-8.[Method]By using a variety of selective media,varieties of starch degrading bacterium were isolated from the sugar cane bagasse form different stages of natural fermentation,then,primary screening and secondary screening were performed.[Result] Starch-degrading strains s2g5-1 and s3g4-8 were screened,and they were identified as Bacillus amyloliquefaciens according to their morphological,physiological,biochemical and molecular characteristics.[Conclusion]The research provided theoretical basis for factory application of bagasse.
文摘目的:验证G2/S期应答相关蛋白1(G2 and S phase-expressed protein 1,GTSE1)在原发性肝癌组织及细胞株中高表达,并探讨其表达下调对肝癌细胞凋亡的影响。方法:采用内置R程序对GEO(Gene Expression Omnibus)数据库中4项原发性肝癌临床研究的基因表达矩阵数据进行分析,筛选肝癌与癌旁组织差异表达基因。采用蛋白质印迹法验证3例肝癌组织和3株肝癌细胞中GTSE1蛋白表达水平。向肝癌Hep-G2和Bel-7402细胞中分别转染GTSE1 siRNA后,采用CCK-8和FCM法分别检测GTSE1表达下调对肝癌细胞增殖、凋亡及5-氟尿嘧啶(5-fluorouracil,5-FU)敏感性的影响,进一步采用蛋白质印迹法检测p53含量及其亚细胞定位,以及Bcl-2家族成员的表达变化。结果:4项研究中原发性肝癌组织表达水平均上调和下调的基因分别为51和146个。其中,上调基因产物GTSE1蛋白在3例临床肝癌组织标本和3株肝癌细胞Hep-G2、Bel-7402和SMMC-7721中均得到进一步验证(P值均<0.001)。siRNA转染后,肝癌Hep-G2和Bel-7402细胞中GTSE1基因沉默效率>70%。与对照组相比,GTSE1表达下调后肝癌Hep-G2和Bel-7402细胞增殖被明显抑制,并随着时间延长,抑制作用逐渐增强(P值均<0.05);GTSE1表达下调后肝癌Hep-G2和Bel-7402细胞周期被阻滞在G1期,同时细胞凋亡率明显升高,且细胞对5-FU的敏感性明显增强(P值均<0.01)。GTSE1表达下调后的肝癌Hep-G2和Bel-7402细胞中总p53及磷酸化P53表达水平均明显上调(P值均<0.05),且细胞核内p53水平升高(P值均<0.01);Bcl-2家族促凋亡成员Bax和Bak表达水平均上调,而抗凋亡成员Bcl-2表达水平下调(P值均<0.05)。结论:下调肝癌中GTSE1蛋白表达能通过活化p53通路促进细胞凋亡,并增加肝癌细胞对化疗药物的敏感性。
