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张辉, 喻莉萍, 李旭, 黄常洪, 胡吉林, 姚飞虹, 李强国. La(Sal)2(Qu)对重组Fas基因酵母促凋亡作用[J]. 中国公共卫生, 2010, 26(11): 1357-1359. DOI: 10.11847/zgggws2010-26-11-07
引用本文: 张辉, 喻莉萍, 李旭, 黄常洪, 胡吉林, 姚飞虹, 李强国. La(Sal)2(Qu)对重组Fas基因酵母促凋亡作用[J]. 中国公共卫生, 2010, 26(11): 1357-1359. DOI: 10.11847/zgggws2010-26-11-07
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ZHANG Hui, YU Li-ping, LI Xu, . Apoptosis induced by La( Sal)2(Qu) in S. pombe with recombinant Fas gene[J]. Chinese Journal of Public Health, 2010, 26(11): 1357-1359. DOI: 10.11847/zgggws2010-26-11-07
Citation: ZHANG Hui, YU Li-ping, LI Xu, . Apoptosis induced by La( Sal)2(Qu) in S. pombe with recombinant Fas gene[J]. Chinese Journal of Public Health, 2010, 26(11): 1357-1359. DOI: 10.11847/zgggws2010-26-11-07
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La(Sal)2(Qu)对重组Fas基因酵母促凋亡作用

Apoptosis induced by La( Sal)2(Qu) in S. pombe with recombinant Fas gene

    摘要
  • 摘要: 目的观察氯化镧水杨酸8-羟基喹啉三元稀土配合物La(C7H5O3)2·(C9H6NO),对重组Fas基因酵母的促凋亡作用。方法抽提Jurkat细胞RNA;逆转录-聚合酶链反应(RT-PCR)制备Jurkat细胞的cDNA;PCR扩增人Fas基因;将人Fas基因克隆到pREP3X-HA质粒中构建pREP3X-HA-Fas穿梭载体,电转化穿梭载体到野生型粟酒裂殖酵母细胞中;应用蛋白印迹法(Western blotting)鉴定蛋白表达产物;应用La(Sal)2(Qu)诱导重组Fas酵母产生凋亡应答。结果经原位末端标记(TUNEL)法显微计数检测发现,对照组细胞膜泡形成、细胞核碎裂和细胞核溶解的细胞各占计数细胞总数的2.80%、3.74%和2.80%,而处理组分别为26.67%、22.86%和13.33%,差异有统计学意义(P<0.01);流式细胞术检测发现,对照组细胞凋亡检出率为1.24%,而处理组为50.17%,差异有统计学意义(P<0.01)。结论 La(Sal)2(Qu)可能通过Fas诱导酵母细胞凋亡。

     

    Abstract: ObjectiveTo observe the apoptosis induced by La(C7H5O3)2·(C9H6NO)in recombinant yeast.Meth-ods Total RNA of Jurkat cells was extracted by TRIZOL reagent.First-strand cDNA of Jurkat cells was synthesized by RT-PCR.The Fas genes were amplified by PCR and cloned into plasmid and formed a relevant shuttle carrier.The pREP3X-HA-Fas shuttle carriers were transformed into wild-type Schizosacharomyces pombe(S.pombe)by electroporation.The active recombination was identified by Western blotting and the apoptosis induced by La(C7H5O3)2·(C9H6NO)was observed.ResultsThe ratio of the bubble of cellular membrane,the fragmentation and caryolysis of cell nucleus induced by La(C7H5O3)2·(C9H6NO)were 26.67%,22.86%,and 13.33%,respectively,detected with terminal deoxynucleotidyl transferase dUTP nick end labeling in treated group with significant differences compared with those of the control group (2.80%,3.74%,and 2.80%,respectively,P<0.01 for all).The apoptosis rate increased in treated group compared with that of the control group detected by flow cytometry(50.17% vs 1.24%,control group vs treated group,P<0.01).ConclusionLa(C7H5O3)2·(C9H6NO)could induce apoptosis by Fas in S.pombe.

     

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