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

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.