Identification by differential display of a mRNA specifically induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) in T84 human colon carcinoma cells.
12-o-Tetradecanoylphorbol-13-Acetate (TPA) down-regulates the expression of the gene responsible for cystic fibrosis (CFTR). To understand the mechanism by which TPA down-regulates CFTR, we decided to study genes specifically induced by this phorbol ester in T84 human colon carcinoma cells, which highly express CFTR, using differential display. Several strategies that allowed us to overcome false-positive reactions in differential displays are described. We have detected different cDNAs obtained from mRNAs specifically induced by TPA. A cDNA fragment corresponding to a mRNA of approximately 2.2 kb was sequenced. Part of this sequence has been reported by others in GenBank and corresponds to a cDNA from a human lung library. The function is unknown and does not have any significant homology with other sequences. It is expressed after two hrs in T84 cells treated with TPA, reaching a maximum response by four hrs. The dose-response curve shows increased mRNA levels starting at 10 ng/ml of TPA and reaching a maximum by 50 ng/ml TPA (10-fold stimulation over control). This mRNA shows a rapid and large response to TPA and it might be involved in the differentiation of T84 cells induced by TPA.
Cafferata
E G
EG
Instituto de Investigaciones Bioquímicas, Fundación Campomar, Buenos Aires, Argentina.
Gonzalez-Guerrico
A M
AM
Pivetta
O H
OH
Santa-Coloma
T A
TA
D001483:Base Sequence; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D003001:Cloning, Molecular; D003110:Colonic Neoplasms / Q000235:genetics* / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D015536:Down-Regulation / Q000187:drug effects; D006801:Humans; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics*; D012334:RNA, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
C505032:CFTR protein, human; D018076:DNA, Complementary; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D013755:Tetradecanoylphorbol Acetate