Cancer Research CTRC-AACR San Antonio Breast Cancer Symposium AACR Conference on Molecular Diagnostics - 2008
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wu, W. K.K.
Right arrow Articles by Cho, C. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wu, W. K.K.
Right arrow Articles by Cho, C. H.
[Cancer Research 65, 5272-5277, June 15, 2005]
© 2005 American Association for Cancer Research

Cell and Tumor Biology

4-(Methylnitrosamino)-1-(3-Pyridyl)-1-Butanone from Cigarette Smoke Stimulates Colon Cancer Growth via ß-Adrenoceptors

William K.K. Wu1, Helen P.S. Wong1, Shi W. Luo1, Kevin Chan2, Fung Y. Huang1, Marco K.C. Hui1, Emily K.Y. Lam1, Vivian Y. Shin1, Yi N. Ye1, Ying H. Yang1 and Chi H. Cho1

1 Department of Pharmacology, Faculty of Medicine, The University of Hong Kong and 2 School of Chinese Medicine, Hong Kong Baptist University, Hong Kong, China

Requests for reprints: Chi H. Cho, Department of Pharmacology, The University of Hong Kong, 21 Sassoon Road, Hong Kong, China. Phone: 852-2819-9250; Fax: 852-2817-0859; E-mail: chcho{at}hkusua.hku.hk.


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
Cigarette smoking is a risk factor for colorectal cancer. It is suggested that 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a tobacco-specific nitrosamine, mediates the carcinogenic action of cigarette smoking by promoting cancer growth. In the present study, the proliferative response of a cultured colon cancer cell line HT-29 to NNK was determined. It was found that NNK dose-dependently stimulated HT-29 cell proliferation. In this regard, the stimulatory action of NNK was abolished by atenolol and ICI 118,551, a ß1- and ß2-selective antagonist, respectively. In addition, cell growth was stimulated by the nonselective adrenergic agonist, noradrenaline, and more effectively by the ß-selective agonist, isoproterenol. The second message cyclic AMP level for ß-adrenoceptor activation was elevated by isoproterenol and NNK treatment. These agents also up-regulated cyclooxygenase-2 expression, cytosolic phospholipase A2 expression, and prostaglandin E2 release. ß2-adrenoceptor blockade with ICI 118,551, in contrast, significantly decreased cyclooxygenase-2 expression, cytosolic phospholipase A2 expression and prostaglandin E2 release induced by NNK and isoproterenol. To conclude, it is proposed that NNK stimulates HT-29 cell proliferation through ß-adrenoceptors, preferentially ß2 receptors. Activation of the ß-adrenoceptors, and the consequent cyclic AMP elevation coupled with the downstream arachidonic acid pathway, is perhaps an important mechanistic cascade in the promotion of colon cancer growth. These findings partly elucidate the carcinogenic actions of cigarette smoke and shed new light on the novel modulatory role of ß-adrenoceptors in the development of colon cancer.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
Cigarette smoking is a putative risk factor for colorectal cancer. Despite the inconsistent association between cigarette smoking and colorectal cancer reported in early studies (1), it was documented that cigarette smoking was related to adenomatous colorectal polyps, which are the precursor to colorectal cancers (2). The risk of larger polyps is also positively associated with a longer duration of smoking (3, 4). In addition, the risk of adenoma remained elevated for up to 10 years after smoking cessation (5). In 2003, Colangelo et al. reported their findings concerning the correlation between smoking and mortality from colorectal cancer in a cohort of 39,299 subjects with an average of 26 years of follow-up, affirming the relationship between cigarette smoking and colorectal cancer (6). Colorectal cancer is the second most common cause of cancer death in the Western world (7). The molecular and cellular events involved in the pathogenesis of smoking-related colorectal cancer, however, remain understudied.

4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a tobacco-specific nitrosamine, has been reported to play a key role in the initiation and promotion of smoking-related malignancy (8). NNK is formed during the processing and curing of tobacco plants (9). It is also believed that NNK could be converted endogenously from nicotine, a major alkaloid in cigarette smoke (10). This tobacco-specific nitrosamine has a high affinity for ß-adrenergic receptors because of its structural resemblance to classical ß-adrenergic agonists (11). In this respect, it was found that ß-adrenoceptor antagonists could reverse the cancer-promoting action of NNK in human cell lines derived from pulmonary adenocarcinomas and pancreatic ductal carcinomas (12, 13). There is also another line of evidence supporting that the pharmacologic action of NNK could be mediated by nicotinic acetylcholine receptors (nAChRs), especially, {alpha}7-nAChR (14). In particular, in small cell lung carcinoma, it was found that NNK could promote cancer growth through activation of Raf-1/mitogen-activated protein kinase/c-myc signaling pathway through {alpha}7-nAChR activation (15). {alpha}-Bungarotoxin, a specific {alpha}7-nAChR antagonist, was able to reverse NNK-induced 5-lipoxygenase and cyclooxygenase-2 (COX-2) protein expressions in SW1116 cells in which activation of the arachidonic acid cascade was implicated in the proliferation of colon cancer cell (14). It is therefore speculated that the cancer-promoting action of NNK is likely mediated through the ß-adrenoceptors and nAChRs in colon cancer cells and its downstream arachidonic acid pathway, especially COX-2 activation in relation to colon cancer growth.

Several investigators have reported the expression of ß-adrenergic receptors in normal colon tissue (16, 17) but their modulatory action on cancer growth and their relationship to smoking-related colorectal cancer are elusive. In the present study, the modulatory roles of NNK in HT-29 colon cancer cell proliferation were clarified. Using selective agonists and antagonists of ß-adrenoceptors, we would like to delineate the differential involvement of these receptors (both ß1- and ß2) and their role in the promoting action of NNK on colon cancer growth.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Reagents and drugs. NNK was purchased from Chemsyn Science Laboratories (Lenexa, KS). Atenolol (ß1-selective antagonists), ICI 118,551 (ß2-selective antagonists), and isoproterenol (ß-selective agonists) were purchased from Sigma (St. Louis, MO). Antibodies for COX-2 and cytosolic phospholipase A2 (cPLA2) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Prostaglandin E2 (PGE2) enzyme immunoassay kit was obtained from R&D Systems (Minneapolis, MN).

Cell culture and viability assays. The human colon cancer cell lines HT-29, SW1116, SW480, and CCD18Co were obtained from the American Type Culture Collection (Manassas, VA) and maintained in RPMI 1640, supplemented with 10% fetal bovine serum, 100 units/mL penicillin and 100 µg/mL streptomycin at 37°C in a humidified atmosphere of 5% CO2 and 95% air. Cell viability was determined by trypan blue exclusion assay. In brief, cells treated with various agents were trypsinized and stained with 0.4% trypan blue. Viable cells were counted with a hemocytometer.

Cell proliferation assay. Cell proliferation was assessed as DNA synthesis. To evaluate DNA synthesis in cells, the incorporation of [3H]thymidine into DNA was determined. Briefly, 5 x 104 cells were seeded for attachment. They were incubated with various substances for 4 hours. In order to study the effects of the antagonists, cells were pretreated with 100 µmol/L atenolol or 50 µmol/L ICI 118,551 for 45 minutes prior to treatment with 100 nmol/L NNK or 100 µmol/L isoproterenol. In the next step, 0.5 µCi of [3H]thymidine was added to each well, and the cells were incubated for a further 4 hours. The final incorporation of [3H]thymidine into cells was measured with a liquid scintillation counter (LS-6500, Beckman Instruments, Inc., Pullerton, CA).

Reverse transcription-PCR for ß1-, ß2-adrenoceptor, epidermal growth factor receptor, cyclooxygenase-2, and ß-actin mRNA. Total RNA was isolated from colon cancer cells using TRIZOL Reagent (Life Technologies BRL, Gaithersburg, MD). Five micrograms of the total RNA was used to generate the first strand of cDNA by reverse transcription (Life Technologies BRL) in accordance with the manufacturer's instructions. Specific primers (Table 1) were designed to screen the expression of ß1-, ß2-adrenoceptors, and epidermal growth factor receptor (EGFR) as well as for semiquantitation of COX-2 mRNA expression. The PCR conditions for the detection of adrenoceptors mRNA expression were as follows: the template cDNA was first denatured at 94°C for 5 minutes. During 32 cycles of amplification, the denaturation step was at 94°C for 1 minute, the annealing step at 54°C for 1 minute and the extension step at 72°C for 1 minute. The final extension step was at 72°C for 7 minutes. The PCR condition for the semiquantitation of COX-2 mRNA expression was as follows: the template cDNA was first denatured at 94°C for 5 minutes. During 30 cycles of amplification, the denaturation step was at 94°C for 1 minute, the annealing step at 58°C for 1 minute and the extension step at 72°C for 1 minute. The final extension step was at 72°C for 7 minutes. The PCR products were electrophoresed on 1.5% (w/v) agarose gels containing 0.5 µg/mL ethidium bromide. Gel photographs were then analyzed semiquantitatively in a multianalyzer (Bio-Rad Laboratories, Hercules, CA).


View this table:
[in this window]
[in a new window]
  		Table 1. Oligonucleotide sequence of primers used for RT-PCR

 
Determination of cyclic AMP. Intracellular cyclic AMP (cAMP) assay was done as described previously (18). Briefly, 1 x 106 cells had been treated with or without NNK at 100 nmol/L or isoproterenol at 100 µmol/L for 30 minutes. The cAMP level was measured by RIA according to the manufacturer's recommendation (Amersham Pharmacia Biotech, Braunschweig, Germany). The cAMP level was expressed as picomoles per milligram of protein.

Western blot. After the same procedures of incubation as described in the cell proliferation assay, protein expressions for COX-2 and cPLA2 were determined by Western blot. Briefly, cells were lysed in radioimmunoprecipitation assay buffer [50 mmol/L Tris-HCl (pH 7.5), 150 mmol/L sodium chloride, 0.5% {alpha}-cholate acid, 0.1% SDS, 2 mmol/L EDTA, 1% Triton X-100, and 10% glycerol], containing 1.0 mmol/L phenylmethylsulfonyl fluoride and 1 µg/mL aprotinin. After sonication for 30 seconds on ice and centrifuging for 20 minutes at 12,000 rpm at 4°C, the supernatant was collected and protein concentration was determined by protein assay kit (Bio-Rad) using bovine serum albumin as standard. Fifty micrograms of protein samples were resolved on SDS-PAGE and transferred to Hybond C nitrocellulose membranes (Amersham Corporation, Arlington Heights, IL). The membrane was probed with COX-2 or cPLA2 antibodies overnight at 4°C and incubated for 1 hour with secondary antibodies conjugated with peroxidase. The signals on the membranes were visualized by enhanced chemiluminescence (Amersham Corporation, Arlington Heights, IL) and exposed on X-ray film (Fuji Photo Film Co., Ltd., Tokyo, Japan). Quantification of the protein bands was carried out by video densitometry (Scan Marker III, Microtek, Carson, CA).

Prostaglandin E2 assay. After the same procedures of incubation as described in the cell proliferation assay, cells were lysed in enzyme immunoassay buffer [50 mmol/L Tris-HCl (pH 7.4), 100 mmol/L NaCl, 1 mmol/L CaCl2, 1 mg/mL D-glucose, and 28 µmol/L indomethacin] for 30 seconds on ice by sonication. Samples were then centrifuged for 15 minutes at 12,000 rpm at 4°C. The supernatant was used for the determination of PGE2 using the PGE2 enzyme immunoassay kit according to the manufacturer's instructions. The optical densities were determined by the MRX microplate reader (Dynex Technologies, Chantilly, VA) at 405 nm.

Statistical analysis. Results are expressed as the mean ± SE of determinations done in triplicate, and statistical comparisons are based on Student's t test or ANOVA. P < 0.05 was considered to be significant.


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
HT-29 and two other colon cell lines expressed ß-adrenoceptors mRNA. As ß-adrenoceptors have been implicated in the mediation of cancer-promoting action on pulmonary and pancreatic cancer cells (12, 13), the mRNA expression of ß1- and ß2-adrenoceptors was screened in three different colon cell lines using RT-PCR method. It was found that mRNA expression of both ß1- and ß2-adrenoceptors was detected in HT-29, SW1116, and SW480 (Fig. 1). Notably, HT-29 strongly expressed mRNA of ß2-adrenoceptor. In contrast, expression of ß1-adrenoceptor was just barely detectable in HT-29. In addition, EGFR mRNA was also highly expressed in these colon cancer cells. Human colon fibroblasts also expressed ß1- and ß2- adrenoceptors and EGFR in a similar pattern.



View larger version (47K):
[in this window]
[in a new window]
  		Figure 1. RT-PCR revealed expression of ß1-, ß2-adrenoceptors, and EGFR mRNA in three colon cancer cell lines (SW1116, HT-29, and SW480). Expression of mRNA of these receptors was also analyzed in human colon fibroblast CCD18Co. M, 50 bp marker; +, ß-actin; –, negative control (PCR product without reverse transcription).

 
NNK increased HT-29 cell proliferation which was reversed by ß-adrenoceptor blockade. To study the effect of NNK on proliferation of colon cancer cells, we examined change in [3H]thymidine incorporation in response to NNK in cultured HT-29 cells. In Fig. 2A, NNK significantly enhanced [3H]thymidine incorporation into HT-29 cells in a concentration-dependent manner. Giving a dose of NNK as low as 100 nmol/L significantly increased HT-29 DNA synthesis by about 21% when compared with the control group. Herein, we chose 100 nmol/L NNK as the working concentration for subsequent experiments in this study.



View larger version (19K):
[in this window]
[in a new window]
  		Figure 2. A, the effects of 4-hour treatment with NNK on HT-29 DNA synthesis. Incubation with NNK increased DNA synthesis in HT-29 cells in a concentration-dependent manner. B, the effects of ß1- and ß2-adrenoceptor blockade with atenolol and ICI 118, 551 on NNK-induced HT-29 cell proliferation. HT-29 cells were pretreated with atenolol or ICI 118,551 before incubation with NNK. Pretreatment with both ß1- and ß2-antagonists reversed the NNK-induced cell proliferation. ß2-Selective antagonist produced a more prominent effect in blocking the NNK-enhanced cell proliferation. C, the effects of ß-selective and nonselective adrenoceptor agonists on HT-29 cell proliferation. Incubation with both isoproterenol or noradrenaline at a dose of 100 µmol/L significantly increased HT-29 cell proliferation. However, treating cells with ß-selective agonist isoproterenol at a dose of 1 µmol/L produced a similar effect, signifying the involvement of ß-adrenoceptors in colon cancer cell growth. *, P < 0.05; **, P < 0.005, significantly different from the untreated control group. ++, P < 0.005, significantly different from the NNK-treated group.

 
To elucidate which receptor isoform mediates the stimulatory action, ß1-selective antagonist atenolol and ß2-selective antagonist ICI 118,551 were used. Both agents completely and significantly dampened NNK-induced HT-29 cell proliferation (Fig. 2B), signifying the involvement of both ß1- and ß2-adrenoceptors in this biological process. ICI 118,551 was more potent than atenolol in blocking the action of NNK on cell proliferation, indicating that the ß2-adrenoceptor might play a dominant role in colon cancer growth. The cell viability in all treatment groups was confirmed to be comparable to that in the control group by trypan blue exclusion assay.

ß-Adrenoceptor agonists increased HT-29 cell proliferation. To further elucidate the role of ß-adrenoceptor in the control of colon cancer cell proliferation, noradrenaline and isoproterenol (nonselective versus ß-selective) were employed to determine the effect of adrenergic stimulation on HT-29 cell proliferation. It was found that both noradrenaline and isoproterenol at a dose of 100 µmol/L significantly increased HT-29 cell proliferation by 20% after a 4-hour incubation (Fig. 2C). However, the ß-selective agonist isoproterenol produced similar promoting effects at a dose of as low as 1 µmol/L which was 100 times less than the nonselective agonist noradrenaline to induce proliferation, indicating that ß-adrenoceptor was largely involved in the promotion of colon cancer cell growth.

Cyclic AMP level was elevated upon NNK and isoproterenol treatment in HT-29. To establish that activation of ß-adrenoceptor in HT-29 cells produced a functional response in cancer cells, intracellular levels of cAMP in HT-29 were measured. One hundred micromolar of isoproterenol produced consistent and significant responses on DNA synthesis and cAMP production in HT-29 cells (Figs. 2 and 3). One hundred nanomolar of NNK provoked similar responses indicating that the tumorigenic action of NNK is much more potent than that of isoproterenol. NNK or isoproterenol incubation at these concentrations for 30 minutes markedly increased cAMP levels by 3- and 4.5-fold, respectively (Fig. 3).



View larger version (11K):
[in this window]
[in a new window]
  		Figure 3. The effects of 30-minute treatment with isoproterenol or NNK on HT-29 intracellular cAMP levels. Incubation with isoproterenol or NNK significantly increased cAMP levels in HT-29 cells showing that the activation of ß-adrenoceptors produced a functional response. **, P < 0.005; ***, P < 0.0005, significantly different from the untreated control group.

 
NNK and isoproterenol up-regulated cyclooxygenase-2, cytosolic phospholipase A2 expression and prostaglandin E2 release which were abolished by ß-adrenoceptor antagonists. Previous studies showed that activation of arachidonic acid cascade by NNK could stimulate cell proliferation in the colon. Nevertheless, whether activation of the cascade was under the control of ß-adrenoceptor in the colon was unknown. To this end, it was found that NNK and isoproterenol significantly increased COX-2 mRNA and protein expression, cPLA2 protein expression, and PGE2 release in HT-29 cells. In contrast, both ß1- and ß2-selective antagonists significantly suppressed COX-2, cPLA2 expression, and PGE2 release in which ß2-selective antagonist produced a more prominent effect (Figs. 4 and 5).



View larger version (30K):
[in this window]
[in a new window]
  		Figure 4. A, expression of COX-2 mRNA in HT-29 cells was increased by NNK and isoproterenol as revealed by RT-PCR analysis. Blockade with ß1- and ß2-selective antagonists reversed the effects. Lane 1, control; lane 2, 100 nmol/L NNK; lane 3, 100 nmol/L NNK + 100 µmol/L atenolol; lane 4, 100 nmol/L NNK + 50 µmol/L ICI 118,551; lane 5, 100 µmol/L isoproterenol; lane 6, 100 µmol/L isoproterenol + 100 µmol/L atenolol; lane 7, 100 µmol/L isoproterenol + 50 µmol/L ICI 118,551; lane 8, 100 µmol/L atenolol; lane 9, 50 µmol/L ICI 118,551. B, Western blot analysis of COX-2 protein expression. *, P < 0.05; ***, P < 0.0005, significantly different from the control group. ++, P < 0.005, +++, P < 0.0005, significantly different from the NNK treated group. #, P < 0.05; ###, P < 0.0005, significantly different from the isoproterenol treated group.

 


View larger version (26K):
[in this window]
[in a new window]
  		Figure 5. A, protein expression of cPLA2 in HT-29 cells was increased by NNK and isoproterenol. Blockade with ß1- and ß2-selective antagonists reversed the effects. B, expression of PGE2 in HT-29 cells was increased by NNK and isoproterenol. Blockade with ß1- and ß2-selective antagonists reversed the effects. *, P < 0.05; **, P < 0.005, ***, P < 0.0005, significantly different from the control group. +++, P < 0.0005, significantly different from the NNK-treated group. ###, P < 0.0005, significantly different from the isoproterenol-treated group.

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
NNK is a highly carcinogenic tobacco-specific nitrosamine but its role in smoking-related colorectal cancer carcinogenesis remained unknown. In the present study, we showed that all screened colon carcinoma cell lines expressed ß1- and ß2-adrenoceptor mRNAs. In addition, NNK was found to stimulate colon cancer HT-29 cell proliferation and the effect could be abolished by ß1- or ß2-antagonists. Treatment with NNK and ß-adrenoceptor agonist also increased the intracellular cAMP level in HT-29 cells. All these findings suggested that ß-adrenoceptors mediated the mitogenic action of NNK via a functional signal transduction pathway.

Previous studies showed that arachidonic acid cascade was activated during colon cancer growth (19–21). Here we report for the first time that NNK and ß-adrenoceptors agonist in HT-29 cells increased cPLA2 protein expression, COX-2 mRNA, and protein expression, as well as PGE2 release which could also be abolished by ß1- or ß2-blockade, suggesting that ß-adrenoceptors are functionally coupled to the arachidonic acid cascade in colon cancer cells. In this respect, it was reported that protein kinase A activated by cAMP binding was able to phosphorylate Src and subsequently activate the Erk1/2 cascade (22). Activation of Erk1/2 has been linked to the up-regulation of COX-2 activity (23). It is therefore possible that the effect of ß-adrenoceptor stimulation by NNK on arachidonic acid cascade activation was signaling through cAMP-dependent Src-mediated pathway. Alternatively, persistently activated ß-adrenoceptors might recruit ß-arrestin, which consequently phosphorylates the downstream protein tyrosine kinases and phospholipase C{gamma} (24, 25). Phospholipase C{gamma} generates the second messenger, diacylglycerol, which is an activator for the protein kinase C{alpha} (26). It has also been shown that protein kinase C{alpha} could regulate the activity of arachidonic acid cascade.

The above findings clearly indicated that the ß-adrenoceptors in fact assumed an important role in the progression of smoking-related colorectal cancer. Our data suggested that the activation of ß-adrenoceptors would functionally result in increased cell proliferation, accompanied with the up-regulation of arachidonic acid cascade. This finding might partially explain the protective effects of nonsteroidal antiinflammatory drugs and COX-2 inhibitors in the treatment and prevention of colorectal cancer in which multiple extracellular mitogenic signals have been reported to converge on the downstream arachidonic acid cascade (14, 27). More noteworthy was that treating HT-29 cells with ß2-antagonist alone in the absence of NNK or ß-adrenoceptor agonist stimulation significantly down-regulated the basal cPLA2 expression, COX-2 expression, along with the concomitant inhibition of cell proliferation. The above observation suggested that the basal activation of arachidonic acid cascade and cell proliferation were actually dependent on the constitutive activity of ß-adrenoceptor-mediated pathway, which was susceptible to ß-adrenoceptor blockade. One of the explanations was that the cancer cells might secrete low levels of epinephrine or norepinephrine to self-stimulate their ß-adrenoceptors in an autocrine or paracrine manner.

Cigarette smoking is a risk factor for colorectal cancer. In this study, we were able to show that the cancer-promoting action of tobacco-specific nitrosamine NNK was ß-adrenoceptor-mediated. The possible involvement of ß-adrenoceptors in colorectal cancer implicated that other factors that increased the levels of endogenous ß-adrenoceptor agonist would promote colorectal cancer progression. In the context of smoking-related colorectal cancer, it was well documented that nicotine could induce secretion of norepinephrine from adrenal glands (28, 29). It is therefore possible that the elevated level of norepinephrine induced by nicotine might actually account for one of the cancer-promoting mechanisms of cigarette smoking.

Activation of ß-adrenoceptors has been reported to stimulate pulmonary and pancreatic carcinoma cell growth as well as colon cancer cell migration (12, 13, 30). To date, there are at least three reports showing a negative relationship between the use of ß-blockers and cancer risk (31–33). In these studies, ß-blockers rather than diuretics or calcium channel blockers for cardiovascular diseases reduced the cancer risk. Pertaining to colorectal cancer, polymorphisms in the ß2- and ß3-adrenoceptor genes are also found to be associated with the cancer risk (34), further substantiating the role of ß-adrenoceptors in the carcinogenesis of colorectal cancer. These findings along with our experimental data might shed new light on the understanding of the carcinogenic mechanism in smoking-related colorectal cancer and possibly open up new chemoprophylactic and therapeutic avenues for the prevention and treatment of cancers.


   Acknowledgments
 
Grant support: Hong Kong Research Grants Council (HKU 7312/04M).

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.


   Footnotes
 
Note: W.K.K. Wu and H.P.S. Wong contributed equally to the paper.

Received 1/21/05. Revised 3/24/05. Accepted 4/ 7/05.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Vineis P, Alavanja M, Buffler P, et al. Tobacco and cancer: recent epidemiological evidence. J Natl Cancer Inst 2004;96:99–106.[Free Full Text]
  2. Terry MB, Neugut AI, Bostick RM, et al. Risk factors for advanced colorectal adenomas: a pooled analysis. Cancer Epidemiol Biomarkers Prev 2002;11:622–9.[Abstract/Free Full Text]
  3. Reid ME, Marshall JR, Roe D, et al. Smoking exposure as a risk factor for prevalent and recurrent colorectal adenomas. Cancer Epidemiol Biomarkers Prev 2003;12:1006–11.[Abstract/Free Full Text]
  4. Giovannucci E, Rimm EB, Stampfer MJ, et al. A prospective study of cigarette smoking and risk of colorectal adenoma and colorectal cancer in U.S. men. J Natl Cancer Inst 1994;86:183–91.[Abstract/Free Full Text]
  5. Chao A, Thun MJ, Jacobs EJ, et al. Cigarette smoking and colorectal cancer mortality in the cancer prevention study II. J Natl Cancer Inst 2000;92:1888–96.[Abstract/Free Full Text]
  6. Colangelo LA, Gapstur SM, Gann PH, Dyer AR. Cigarette smoking and colorectal carcinoma mortality in a cohort with long-term follow-up. Cancer 2004;100:288–93.[CrossRef][Medline]
  7. CRC CancerStats. London: Cancer Research Campaign; 1999.
  8. Wu WK, Cho CH. The pharmacological actions of nicotine on the gastrointestinal tract. J Pharmacol Sci 2004;94:348–58.
  9. Adams JD, Lee SJ, Vinchkoski N, Castonguay A, Hoffmann D. On the formation of the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone during smoking. Cancer Lett 1983;17:339–46.[CrossRef][Medline]
  10. Hecht SS, Hochalter JB, Villalta PW, Murphy SE. 2'-Hydroxylation of nicotine by cytochrome P450 2A6 and human liver microsomes: formation of a lung carcinogen precursor. Proc Natl Acad Sci U S A 2000;97:12493–7.[Abstract/Free Full Text]
  11. Schuller HM, Tithof PK, Williams M, Plummer H III. The tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone is a ß-adrenergic agonist and stimulates DNA synthesis in lung adenocarcinoma via ß-adrenergic receptor-mediated release of arachidonic acid. Cancer Res 1999;59:4510–5.[Abstract/Free Full Text]
  12. Schuller HM, Cole B. Regulation of cell proliferation by ß-adrenergic receptors in a human lung adenocarcinoma cell line. Carcinogenesis 1989;10:1753–5.[Abstract/Free Full Text]
  13. Weddle DL, Tithoff P, Williams M, Schuller HM. ß-Adrenergic growth regulation of human cancer cell lines derived from pancreatic ductal carcinomas. Carcinogenesis 2001;22:473–9.[Abstract/Free Full Text]
  14. Ye YN, Liu ES, Shin VY, Wu WK, Cho CH. The modulating role of nuclear factor-{kappa}B in the action of {alpha}7-nicotinic acetylcholine receptor and cross-talk between 5-lipoxygenase and cyclooxygenase-2 in colon cancer growth induced by 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone. J Pharmacol Exp Ther 2004;311:123–30.[Abstract/Free Full Text]
  15. Jull BA, Plummer HK III, Schuller HM. Nicotinic receptor-mediated activation by the tobacco-specific nitrosamine NNK of a Raf-1/MAP kinase pathway, resulting in phosphorylation of c-myc in human small cell lung carcinoma cells and pulmonary neuroendocrine cells. J Cancer Res Clin Oncol 2001;127:707–17.[Medline]
  16. Schultheiss G, Diener M. Adrenoceptor-mediated secretion across the rat colonic epithelium. Eur J Pharmacol 2000;403:251–8.[CrossRef][Medline]
  17. Kennedy MF, Tutton PJ, Barkla DH. Adrenergic factors involved in the control of crypt cell proliferation in jejunum and descending colon of mouse. Clin Exp Pharmacol Physiol 1983;10:577–86.[Medline]
  18. Naro F, Sette C, Vicini E, et al. Involvement of type 4 cAMP-phosphodiesterase in the myogenic differentiation of L6 cells. Mol Biol Cell 1999;10:4355–67.[Abstract/Free Full Text]
  19. Koehne CH, Dubois RN. COX-2 inhibition and colorectal cancer. Semin Oncol 2004;31:12–21.
  20. Ferrandez A, Prescott S, Burt RW. COX-2 and colorectal cancer. Curr Pharm Des 2003;9:2229–51.[CrossRef][Medline]
  21. Mann JR, DuBois RN. Cyclooxygenase-2 and gastrointestinal cancer. Cancer J 2004;10:145–52.[Medline]
  22. Fredriksson JM, Lindquist JM, Bronnikov GE, Nedergaard J. Norepinephrine induces vascular endothelial growth factor gene expression in brown adipocytes through a ß-adrenoreceptor/cAMP/protein kinase A pathway involving Src but independently of Erk1/2. J Biol Chem 2000;275:13802–11.[Abstract/Free Full Text]
  23. Brambilla R, Neary JT, Cattabeni F, et al. Induction of COX-2 and reactive gliosis by P2Y receptors in rat cortical astrocytes is dependent on ERK1/2 but independent of calcium signalling. J Neurochem 2002;83:1285–96.[CrossRef][Medline]
  24. Luttrell LM, Ferguson SS, Daaka Y, et al. ß-Arrestin-dependent formation of ß2 adrenergic receptor-Src protein kinase complexes. Science 1999;283:655–61.[Abstract/Free Full Text]
  25. Cook SJ, Wakelam MJ. Epidermal growth factor increases sn-1,2-diacylglycerol levels and activates phospholipase D-catalysed phosphatidylcholine breakdown in Swiss 3T3 cells in the absence of inositol-lipid hydrolysis. Biochem J 1992;285:247–53.
  26. Kim JH, Kim SW, Jung PJ, Yon C, Kim SC, Han JS. Phosphatidylcholine-specific phospholipase C and RhoA are involved in the thyrotropin-induced activation of phospholipase D in FRTL-5 thyroid cells. Mol Cells 2002;14:272–80.[Medline]
  27. Rice PL, Washington M, Schleman S, Beard KS, Driggers LJ, Ahnen DJ. Sulindac sulfide inhibits epidermal growth factor-induced phosphorylation of extracellular-regulated kinase 1/2 and Bad in human colon cancer cells. Cancer Res 2003;63:616–20.[Abstract/Free Full Text]
  28. Miele E. The nicotinic stimulation of the cat adrenal medulla. Arch Int Pharmacodyn Ther 1969;179:343–51.[Medline]
  29. Narita S, Shibata O, Waki S, Egashira K, Suzuki T. Adrenaline and noradrenaline secretion of the adrenal gland in conscious dogs in response to pilocarpine and nicotine. Tohoku J Exp Med 1973;110:303–5.[Medline]
  30. Masur K, Niggemann B, Zanker KS, Entschladen F. Norepinephrine-induced migration of SW 480 colon carcinoma cells is inhibited by ß-blockers. Cancer Res 2001;61:2866–9.[Abstract/Free Full Text]
  31. Jick H, Jick S, Derby LE, Vasilakis C, Myers MW, Meier CR. Calcium-channel blockers and risk of cancer. Lancet 1997;349:525–8.[CrossRef][Medline]
  32. Pahor M, Guralnik JM, Salive ME, Corti MC, Carbonin P, Havlik RJ. Do calcium channel blockers increase the risk of cancer? Am J Hypertens 1996;9:695–9.[CrossRef][Medline]
  33. Algazi M, Plu-Bureau G, Flahault A, Dondon MG, Le MG. Could treatments with ß-blockers be associated with a reduction in cancer risk? Rev Epidemiol Sante Publique 2004;52:53–65.[Medline]
  34. Takezaki T, Hamajima N, Matsuo K, et al. Association of polymorphisms in the ß-2 and ß-3 adrenoceptor genes with risk of colorectal cancer in Japanese. Int J Clin Oncol 2001;6:117–22.[CrossRef][Medline]



This article has been cited by other articles:


Home page
 J. Pharmacol. Exp. Ther.Home page
X. Liu, W. K. K. Wu, L. Yu, Z. J. Li, J. J. Y. Sung, S. T. Zhang, and C. H. Cho
Epidermal Growth Factor-Induced Esophageal Cancer Cell Proliferation Requires Transactivation of {beta}-Adrenoceptors
J. Pharmacol. Exp. Ther., July 1, 2008; 326(1): 69 - 75.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
L. Mastrangelo, A. Cassidy, F. Mulholland, W. Wang, and Y. Bao
Serotonin Receptors, Novel Targets of Sulforaphane Identified by Proteomic Analysis in Caco-2 Cells
Cancer Res., July 1, 2008; 68(13): 5487 - 5491.
[Abstract] [Full Text] [PDF]

Home page
 Toxicol SciHome page
H. P. S. Wong, L. Yu, E. K. Y. Lam, E. K. K. Tai, W. K. K. Wu, and C.-H. Cho
Nicotine Promotes Colon Tumor Growth and Angiogenesis through {beta}-Adrenergic Activation
Toxicol. Sci., June 1, 2007; 97(2): 279 - 287.
[Abstract] [Full Text] [PDF]

Home page
 Toxicol SciHome page
V. Y. Shin, W. K. K. Wu, K. M. Chu, M. W. L. Koo, H. P. S. Wong, E. K. Y. Lam, E. K. K. Tai, and C. H. Cho
Functional Role of {beta}-Adrenergic Receptors in the Mitogenic Action of Nicotine on Gastric Cancer Cells
Toxicol. Sci., March 1, 2007; 96(1): 21 - 29.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wu, W. K.K.
Right arrow Articles by Cho, C. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wu, W. K.K.
Right arrow Articles by Cho, C. H.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online