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Tytuł oryginału: Diminished expression of the type II receptor for TGFá (TGFáRII) in T lymphocytes from patients with sezary syndrome is not due to mutations in the receptor's poly-A tract: limitations of the standard RT-PCR in cDNA sequence analysis of homopolymeric base stretches.
Autorzy: Zhang Qian, Capocasale Renold J., Fox Floyd E., Bedian Vahe, Vonderheid Eric C., Rook Alain, Moore Jonni S., Nowell Peter C., Haines Dale S., Wasik Mariusz A.
Źródło: Arch. Immunol. Ther. Exp. 2002: 50 (6) s.421-429, il., tab., bibliogr. 26 poz.
Sygnatura GBL: 304,223

Hasła klasyfikacyjne GBL:
  • genetyka
  • hematologia

    Typ dokumentu:
  • praca kliniczna
  • tytuł obcojęzyczny

    Wskaźnik treści:
  • ludzie

    Streszczenie angielskie: Perihperal blood lymphocytes from patients with Sezary syndrome (SzS) frequently demonstrate decreased surface expression of transforming growth factor á receptor II (TGFáRII). The mechanism of this low TGFáRII expression remains unknown. Because mutations within the poly-A tract of the TGFáRII sequence (nucleotides 709 - 718) were shown to result in diminished TGFáRII expression in other types of malignant tumors, we examined the sequence of the TGFáRII poly-A tract in two SzS-derived cell lines and in peripheral blood SzS cells from 17 SzS patients and 4 control, healthy individuals using DNA sequencing and single-stranded conformation polymorphism (SSCP) analysis. A standard bidirectional, automated sequence analysis of the RT-PCR-generated cDNA TGFáRII fragment showed a heterogenous population of the normal length, 10-, with admixed, shortened, 9-base poly-A stretches. Surprisingly, this mixture was present not only in the cells from 5 SzS patients and 2 SzS cell lines, but also in cells from 2 healthy control individuals. Importantly, the proportion of the shortened, 9-base fragments was markedly reduced or practically eliminated when the procedure was modified by usage of high-fidelity DNA polymerase, labeled primers and/or cloned RT-PCR products, which indicates that the presence of the shortened, 9-base fragments represented a procedural phenomenon rather than a true deletional mutation within an allele of the TGFáRII gene. Accordingly, SSCP analysis of genomic DNA did not reveal any mutations within the poly-A tract-containing region. These results indicate ...

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