Journal
LIFE SCIENCES
Volume 71, Issue 21, Pages 2543-2552Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/S0024-3205(02)02082-9
Keywords
HT 29 cell line; vasoactive intestinal peptide (VIP); stearyl-Nle(17)-neurotensin(6-11)VIP(7-28) (SNH); homeobox genes
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A lipophilic analog of vasoactive intestinal peptide (VIP), stearyl-Nle(17)-neurotensin(6-11)VIP(7-28) (SNE), that inhibited lung cancer growth, has been previously described. The mechanism of SNH inhibition of cancer growth is still being elucidated. The present study examined the effects of SNH on homeobox genes in the colon cancer cell line HT 29 that expresses VIP receptors. Homeobox genes contain a characteristic DNA sequence, coding for a stretch of 61 amino acid homeodomain that binds specific DNA motifs. While the HOX gene family contains a single homeodomain, the POU gene family contains an additional DNA binding homeodomain. HT 29 cells were incubated with SNH; RNA was extracted and subjected to reverse-transcription-polymerase chain reaction (RTPCR) with primers that matched the conserved area of the various HOX or POU genes. The PCR products that were altered by SNE treatment were sequenced. Three candidate SNH-responsive genes, the HOX A4, the HOX 135 and the PUO V transcription factor I (Oct-3) were identified. Semi-quantitative RT-PCR with specific primers confirmed the increase in HOX A4 and the decrease in Oct-3 expression levels following SNH treatment. Thus, the HOX A4 and the Oct-3 homeobox genes may partially mediate SNH activity on cancer cells. (C) 2002 Elsevier Science Inc. All rights reserved.
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