Detection of MRP1 mRNA in human tumors and tumor cell lines by in situ RT-PCR

Lorella Pascolo, Cristina Fernetti, Doroti Pirulli, Samanta Bogoni, Maria Victoria Garcia-Mediavilla, Andrea Spanò, Daniela Puzzer, Claudio Tiribelli, Antonio Amoroso, Sergio Crovella

Research output: Contribution to journalArticle

Abstract

The detection of the multridrug resistance-associated proteins is becoming increasingly important in assessing tumor sensitivity to treatment. In this work we describe a new, rapid, sensitive, and robust method for the detection of MRP1 expression based on direct RT-in situ-PCR technology and fluorochrome-modified (dCTP(Cy3)) nucleotides. MRP1 expression was found in both placenta (BeWo) and liver (Hep G2)-derived tumor cell line as well as in small cell lung carcinoma. In liver-derived cells, MRP1 expression was detected by RT-in situ-PCR but not by in situ hybridization, suggesting a higher sensitivity of in situ amplification for the low level of expression in Hep G2 cells. RT solution PCR confirmed the presence of MRP1 in BeWo and Hep G2 cells, although the level of the gene expression was lower in liver cells. This method represents a viable alternative to conventional immunohistochemistry, and may be useful in the evaluation of MRP1 expression in different tissue or cell lines. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)466-471
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume275
Issue number2
DOIs
Publication statusPublished - Aug 28 2000

Keywords

  • BeWo cells
  • Hep G2 cells
  • Human placenta
  • In situ hybridization
  • In situ PCR
  • MRP1

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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    Pascolo, L., Fernetti, C., Pirulli, D., Bogoni, S., Garcia-Mediavilla, M. V., Spanò, A., Puzzer, D., Tiribelli, C., Amoroso, A., & Crovella, S. (2000). Detection of MRP1 mRNA in human tumors and tumor cell lines by in situ RT-PCR. Biochemical and Biophysical Research Communications, 275(2), 466-471. https://doi.org/10.1006/bbrc.2000.3339