The NK-1 receptor is expressed in human primary gastric and colon adenocarcinomas and is involved in the antitumor action of L-733,060 and the mitogenic action of substance P on human gastrointestinal cancer cell lines.
OBJECTIVE
It has been demonstrated that substance P (SP) and neurokinin-1 (NK-1) receptor antagonist L-733,060 induces cell proliferation and inhibition, respectively, in several human cancer cell lines. At present, it is unknown whether such actions are exerted on human gastric and colon adenocarcinomas. We carried out an in vitro study of the growth-inhibitory capacity of L-733,060 against human gastric and colon adenocarcinomas.


METHODS
A coulter counter was used to determine viable cell numbers followed by application of the tetrazolium compound MTS. Immunoblot analysis was used to determine the NK-1 receptors and the DAPI method was applied to demonstrate apoptosis. Immunohistochemistry was used to demonstrate NK-1 receptors in primary human gastric and colon adenocarcinomas.


RESULTS
We observed the presence of several NK-1 receptor isoforms in human gastric and colon adenocarcinomas. Nanomolar concentrations of SP increased the growth of both cell lines and micromolar concentrations of L-733,060 inhibited the growth of such cell lines, with and without previous administration of SP. L-733,060 inhibited the growth of the 23132/87 and SW-403 cell lines in a dose-dependent manner. After administration of L-733,060, apoptosis was observed in both cell lines. In both human primary gastric and colon adenocarcinomas, a high density of NK-1 receptors was observed. Immunoreactivity, showing a diffuse cytoplasmic staining, was observed in the epithelial cells of normal and tumor glands and in numerous stromal elements.


CONCLUSIONS
We demonstrated that NK-1 receptors were expressed in 23132/37 and SW-403 cell lines and in human primary gastric and colon adenocarcinomas, that SP is a mitogen and that the antitumor action of L-733,060 on both human cell lines occurs through the NK-1 receptor. Data also indicate that the cell death observed is produced by apoptosis. These data suggest that the NK-1 receptor is a new and promising target in the treatment of human gastrointestinal adenocarcinomas.
Rosso
Marisa
M
Virgen del Rocío University Children's Hospital, Sevilla, Spain.
Robles-Frías
María Jose
MJ
Coveñas
Rafael
R
Salinas-Martín
Manuel Vicente
MV
Muñoz
Miguel
M
D000230:Adenocarcinoma / Q000737:chemistry* / Q000188:drug therapy; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000737:chemistry* / Q000188:drug therapy / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007150:Immunohistochemistry; D008934:Mitogens / Q000494:pharmacology*; D064729:Neurokinin-1 Receptor Antagonists; D010880:Piperidines / Q000494:pharmacology*; D018040:Receptors, Neurokinin-1 / Q000032:analysis* / Q000502:physiology; D013274:Stomach Neoplasms / Q000737:chemistry* / Q000188:drug therapy / Q000473:pathology; D013373:Substance P / Q000494:pharmacology*
D000970:Antineoplastic Agents; D008934:Mitogens; D064729:Neurokinin-1 Receptor Antagonists; D010880:Piperidines; D018040:Receptors, Neurokinin-1; C103788:3-((3,5-bis(trifluoromethyl)phenyl)methyloxy)-2-phenylpiperidine; D013373:Substance P