title: An alternatively spliced survivin variant is positively regulated by p53 and sensitizes leukemia cells to chemotherapy.
abstract: Survivin is a unique member of the inhibitor of apoptosis protein family, and its expression is regulated by p53. Recent identification of several functionally divergent survivin variants augments the complexity of survivin action as well as its regulation. Here we report that survivin-2B (retaining a part of intron 2 as a cryptic exon) is positively regulated by p53, and its overexpression plays a role in sensitizing leukemia cells to chemotherapeutic drug doxorubicin. Doxorubicin treatment activated p53, downregulated survivin and survivin-DeltaEx3 but upregulated survivin-2B in EU-3, an acute lymphocytic leukemia (ALL) cell line with wild-type (wt)-p53 phenotype. In contrast, doxorubicin treatment failed to induce these alterations in EU-6 cells, a mutant-p53 ALL cell line. To specify the role of wt-p53 in regulating survivin and its variants, a temperature-sensitive p53 mutant plasmid p53-143 was transfected into EU-4, a p53-null ALL cell line, to establish a subline EU-4/p53-143. When EU-4/p53-143 cell culture was shifted from 37.5 degrees C to the wt-p53-permissive temperature (32.5 degrees C), the expression of survivin and survivin-DeltaEx3 was decreased whereas survivin-2B expression was increased, confirming the distinct regulatory effect of p53 on survivin and its variants. To clarify the role of survivin-2B in the process of apoptosis, survivin-2B cDNA was cloned into pcDNA3HA vector and transfected into EU-4 cells. Enforced expression of survivin-2B in EU-4 cells inhibited cell growth and sensitized these cells to doxorubicin-induced apoptosis. These results suggest that survivin-2B variant is a proapoptotic factor and its expression is upregulated by p53.
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Ningxi
authors.initials: N
authors.lastname: Zhu
authors.forename: Lubing
authors.initials: L
authors.lastname: Gu
authors.forename: Harry W
authors.initials: HW
authors.lastname: Findley
authors.forename: Fengzhi
authors.initials: F
authors.lastname: Li
authors.forename: Muxiang
authors.initials: M
authors.lastname: Zhou
mesh_terms: D017398:Alternative Splicing; D000970:Antineoplastic Agents / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D001483:Base Sequence; D002455:Cell Division; D002460:Cell Line; D017931:DNA Primers; D004317:Doxorubicin / Q000494:pharmacology; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D007938:Leukemia / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D009363:Neoplasm Proteins; D000077022:Survivin; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000502:physiology*; D015854:Up-Regulation
chemical_list: D000970:Antineoplastic Agents; C107489:BIRC5 protein, human; D017931:DNA Primers; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D000077022:Survivin; D016159:Tumor Suppressor Protein p53; D004317:Doxorubicin