CCK2 receptor expression transforms non-tumorigenic human NCM356 colonic epithelial cells into tumor forming cells.
Expression of gastrin and cholecystokinin 2 (CCK(2)) receptor splice variants (CCK(2)R and CCK(2i4sv)R) are upregulated in human colonic adenomas where they are thought to contribute to tumor growth and progression. To determine the effects of ectopic CCK(2) receptor variant expression on colonic epithelial cell growth in vitro and in vivo, we employed the non-tumorigenic colonic epithelial cell line, NCM356. Receptor expression was induced using a retroviral expression vector containing cDNAs for either CCK(2i4sv)R or CCK(2)R. RT-PCR and intracellular Ca(2+) ([Ca(2+)](i)) imaging of RIE/CCK(2)R cells treated with conditioned media (CM) from NCM356 revealed that NCM356 cells express gastrin mRNA and secrete endogenous, biologically active peptide. NCM356 cells expressing either CCK(2)R or CCK(2i4sv)R (71 and 81 fmol/mg, respectively) grew faster in vitro, and exhibited an increase in basal levels of phosphorylated ERK (pERK), compared with vector. CCK(2) receptor selective antagonist, YM022, partially inhibited the growth of both receptor-expressing NCM356 cells, but not the control cells. Inhibitors of mitogen activated protein kinase pathway (MEK/ERK) or protein kinase C (PKC) isozymes partially inhibited the elevated levels of basal pERK and in vitro growth of receptor-expressing cells. Vector-NCM356 cells did not form tumors in nude mice, whereas, either CCK(2) receptor-expressing cells formed large tumors. Autocrine activation CCK(2) receptor variants are sufficient to increase in vitro growth and tumorigenicity of non-transformed NCM356 colon epithelial cells through a pathway involving PKC and the MEK/ERK axis. These findings support the hypothesis that expression of gastrin and its receptors in human colonic adenomas contributes to tumor growth and progression.
Chao
Celia
C
Department of Surgery, University of Texas Medical Branch, Galveston, Texas, USA.
Han
Xueliang
X
Ives
Kirk
K
Park
Jeseong
J
Kolokoltsov
Andrey A
AA
Davey
Robert A
RA
Moyer
Mary P
MP
Hellmich
Mark R
MR
D000236:Adenoma / Q000473:pathology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002277:Carcinoma / Q000235:genetics; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division / Q000235:genetics; D003106:Colon / Q000378:metabolism / Q000473:pathology / Q000502:physiology*; D015179:Colorectal Neoplasms / Q000235:genetics / Q000473:pathology*; D017931:DNA Primers; D018450:Disease Progression; D005755:Gastrins / Q000235:genetics / Q000378:metabolism; D014644:Genetic Variation; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism / Q000473:pathology / Q000502:physiology*; D051379:Mice; D009154:Mutation; D009367:Neoplasm Staging; D043887:Receptor, Cholecystokinin B / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D005755:Gastrins; D043887:Receptor, Cholecystokinin B; D002118:Calcium
J Clin Invest. 2000 Aug;106(4):533-9
10953028
Ann Intern Med. 1989 Nov 1;111(9):738-42
2802432
J Biol Chem. 2005 Sep 30;280(39):33368-73
16079138
Int J Cancer. 1999 May 5;81(3):376-86
10209952
Ann N Y Acad Sci. 2004 Apr;1014:97-109
15153424
Digestion. 1999;60(3):210-5
10343134
Cancer Lett. 2007 Jul 8;252(1):19-35
17188424
Gastroenterology. 1993 Jul;105(1):22-30
8514038
Gastroenterology. 1998 Aug;115(2):275-80
9679032
Proc Natl Acad Sci U S A. 1997 Jan 21;94(2):751-6
9012857
Biochim Biophys Acta. 2004 Jul 6;1704(1):1-10
15238241
Cell. 2000 Jan 7;100(1):57-70
10647931
Pharmacol Rev. 1999 Dec;51(4):745-81
10581329
Oncogene. 2007 Feb 15;26(7):1013-9
16909104
Curr Opin Pharmacol. 2007 Dec;7(6):583-92
17997137
Cell. 1996 Oct 18;87(2):159-70
8861899
Nat Rev Genet. 2002 Jun;3(6):415-28
12042769
J Surg Res. 2005 Dec;129(2):313-21
16051276
Gut. 2000 Dec;47(6):820-4
11076881
Gut. 1997 Sep;41(3):330-2
9378387
Cancer Lett. 2005 May 10;222(1):95-105
15837546
Clin Cancer Res. 2000 Dec;6(12):4719-24
11156225
FEBS Lett. 2001 May 4;496(1):25-30
11343700
Cardiovasc Hematol Disord Drug Targets. 2006 Jun;6(2):85-100
16787194
Nat Rev Mol Cell Biol. 2008 Jul;9(7):517-31
18568040
J Biol Chem. 2000 Oct 13;275(41):32122-8
10913157
Am J Surg. 1995 Feb;169(2):190-5; discussion 195-6
7840378
Am J Physiol Gastrointest Liver Physiol. 2003 Jan;284(1):G75-84
12488236
J Biol Chem. 2002 Dec 13;277(50):48755-63
12239223
Nat Rev Cancer. 2001 Oct;1(1):55-67
11900252
Annu Rev Physiol. 2001;63:119-39
11181951
Cancer Res. 2001 Jan 15;61(2):625-31
11212260
Am J Physiol. 1997 Oct;273(4):G891-8
9357832
Br J Cancer. 2002 Aug 27;87(5):567-73
12189558
J Virol. 2005 Jan;79(2):756-63
15613303
Am J Physiol. 1999 Jun;276(6):G1363-72
10362639
Br J Cancer. 2006 Apr 24;94(8):1107-15
16622436