Multiple Gene Blotting to Screen for Cytokine RNA Expression: Application toEndothelioma Cells

摘要

BACKGROUND: Because normal and tumor cells can secrete different cytokines, ithas been a matter of educated guesswork to select which factors to study. Nevertheless, many can be missed, so we have developed a method to screen for unanticipated cytokine expression. EXPERIMENTAL DESIGN: Amplification products from polymerase chain reactions (PCR) of different cytokine genes were spotted onto nylon membranes, and probed with radioactive cDNAs made from mouse endothelioma cells. RESULTS: The presence of interleukin-3 was detected, and confirmed by conventional Northern blotting and polymerase chain reaction (PCR). CONCLUSIONS: This method may be useful to survey cells for the expression of cytokine genes.