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[Cancer Research 63, 8549-8553, December 1, 2003]
© 2003 American Association for Cancer Research

Regular Articles

Polymorphism of the Cyclin D1 Gene, CCND1, and Risk for Incident Sporadic Colorectal Adenomas

Ryan C. Lewis1, Roberd M. Bostick12, Dawen Xie1, Zonglin Deng1, Michael J. Wargovich3, Michael F. Fina4, Walter M. Roufail5 and Kim R. Geisinger6

1 Department of Epidemiology and Biostatistics, Norman J. Arnold School of Public Health, University of South Carolina, Columbia, South Carolina;
2 Departments of Family and Preventive Medicine and
3 Pathology and Microbiology, School of Medicine, University of South Carolina, Columbia, South Carolina;
4 Forsyth Medical Center, Piedmont Gastroenterology Specialists, Winston-Salem, North Carolina; and
5 Departments of Medicine and
6 Pathology, Wake Forest University, School of Medicine, Winston-Salem, North Carolina


   ABSTRACT
Top
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Cyclin D1, encoded by the CCND1 gene and activated by the adenomatous polyposis coli-ß-catenin-T-cell factor/lymphoid enhancing factor pathway, induces G1 to S-phase cell cycle transition, promoting cell proliferation. A recently described codon 242, exon 4, G to A single nucleotide polymorphism (A870G) produces a longer half-life cyclin D1. To investigate whether CCND1 genotype influences risk for colorectal adenoma, we genotyped CCND1 by PCR/RFLP on 161 incident sporadic adenoma cases and 213 controls ages 30–74 years in a North Carolina colonoscopy-based case-control study. At least one polymorphic A allele was found in 68% of cases and 60% of controls. Having an A allele was associated with increased risk for adenoma: the age- and sex-adjusted odds ratio (OR) was 1.5 [95% confidence interval (CI) 1.0–2.4], a finding that was stronger for those whose adenomas were multiple (OR 2.9, 95% CI 1.4–6.0), larger (>=1 cm; OR 2.4, 95% CI 1.2–4.8), had moderate to severe dysplasia (OR 2.1, 95% CI 1.1–3.8), or were in the right side of the colon (OR 3.6, 95% CI 1.3–10.0). Joint risk factor multivariate analyses revealed stronger positive associations among those who were older (>57 years; OR 2.8, 95% CI 1.4–5.5), male (OR 2.8, 95% CI 1.3–5.7), currently smoked (OR 2.7, 95% CI 1.3–5.7), or currently drank alcohol (OR 2.2, 95% CI 1.2–4.2) if they had an A allele and stronger inverse associations among those who used nonsteroidal anti-inflammatory drugs (OR 0.4, 95% CI 0.2–0.9) or had higher calcium intakes (OR 0.4, 95% CI 0.2–0.9) if they had no A allele. These data support the hypothesis that the CCND1 A870G polymorphism may increase risk for colorectal neoplasms.


   INTRODUCTION
Top
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
In the United States, colorectal cancer is currently the third most common cancer, with an estimated 147,500 new cases expected to be diagnosed in 2003 (1) . Ecologic and migrant studies indicate that environmental factors, dietary preferences, and other lifestyle factors greatly influence colorectal cancer risk and account for the marked 10-fold difference in incidence rates between developed and underdeveloped countries (2 , 3) . Data from numerous analytical observational studies (4 , 5) indicate increased risk for colorectal cancer with high calorie/fat diets, red meat intake, physical inactivity, large body mass, smoking, alcohol consumption, and certain occupational exposures. Likewise, lower risk has been frequently associated with diets high in fiber, fruits and vegetables, calcium, and antioxidants, as well as usage of NSAIDs.7 Still, although various environmental factors clearly play a role in colorectal cancer etiology, they do not appear to be the sole determinants.

Sporadic cancers, those for which no traceable pattern of inheritance can be assigned, represent the majority (70%) of all colorectal cancers. A small portion (~5%) of cases occurs in people with either inflammatory bowel diseases or well-defined highly penetrant, autosomal dominantly inherited genetic predispositions such as familial adenomatous polyposis and HNPCC (6 , 7) . The remaining cases occur in individuals with a positive family history, which is associated with a 2–3-fold greater risk for colorectal cancer. Familial adenomatous polyposis results from a loss of function of the APC gene, and HNPCC arises from a loss of function of one of several MMR genes. Silencing of these same genes (i.e., APC and MMR) through somatic mutations or epigenetic mechanisms account for ~80 and 15% of sporadic colorectal cancers, respectively.

Mutations in these pathways are likely to have downstream effects on oncoproteins such as cyclin D1. Regulated by the normal APC-ß-catenin-TCF/LEF pathway, the cyclin D1 protein is involved in cell cycle transition from G1 to S phase (8 , 9) . Therefore, mutations in this pathway are likely to increase expression of cyclin D1 and possibly increase cell proliferation, which may ultimately contribute to the development of colorectal cancer (10 , 11) . Overexpression of cyclin D1 has also been hypothesized to modify the effect of MMR mutations. Although the mechanism is not known, it is suggested that increased cyclin D1 expression contributes to increased microsatellite instability, hence influencing the age of onset of HNPCCs (12 , 13) . In addition, injection of antisense cyclin D1 cDNA was found to suppress growth of cancer cells in nude mice (14) . These findings suggest that cyclin D1 may be involved in multiple pathways of colorectal carcinogenesis.

Cyclin D1 is commonly overexpressed in a variety of epithelial cancers, including colorectal carcinomas (15) . Elevated levels of cyclin D1 have been observed in 30% of human adenocarcinomas and adenomatous polyps (adenomas) of the colon, as well as in 40% of colorectal cancers (16 , 17) . Another study found that both mRNA and protein levels of cyclin D1 were elevated in 50% of colon carcinomas (18) . However, the increased cyclin D1 expression is not the result of gene amplification (8) .

A polymorphic germ-line mutation in the CCND1 gene, which encodes for cyclin D1, was recently reported and may be involved in human carcinogenesis (19) . This particular polymorphism, a G to A mutation located at codon 242 on exon 4 among a splice donor site, additionally enhances CCND1’s alternate splice capability (19) . Both alleles are capable of yielding two fully functional distinct transcripts, a full ‘transcript a’ and a truncated ‘transcript b’; however, the dominant A allele is more closely associated with the truncated transcript, and the G allele tends to produce mostly the full transcript (19 , 20) . The truncated transcript yields a cyclin D1 protein with a longer half-life, thus possibly allowing an over accumulation of cyclin D1 in the cell, in turn, promoting increased cell proliferation (19 , 20) , and thereby increasing the risk for developing colorectal cancer.

There have been few human studies on the association of CCND1 polymorphisms and risk for colorectal cancer. Of five case-control studies, two found significant positive associations for genotype and an increased risk for colorectal cancer (21 , 22) , but three did not (12 , 13 , 23) . However, two of the latter three studies found a younger age of cancer onset with the A allele (12) and the cyclin D1 truncated transcript (13) , whereas the third found a significant positive association with decreased survival time and overexpression of cyclin D1 (23) .

Thus, it is evident from these studies and from cyclin D1’s association with both the APC and MMR colon carcinogenesis pathways that the A/G polymorphism of cyclin D1 may be important in risk for colorectal neoplasms. However, there are no known published epidemiological studies that have evaluated CCND1 genotypes and risk for incident sporadic colorectal adenomatous polyps or levels of epithelial cell proliferation in normal appearing colorectal mucosa and none that have investigated potential interactions between dietary and other environmental risk factors and CCND1 genotypes and risk for colorectal neoplasms. Herein we report a molecular epidemiological study to investigate the association between CCND1 genotypes and risk for colorectal neoplasia, whether the association differs according to adenoma characteristics, whether CCND1 genotype interacts with environmental exposures to modify risk for incident sporadic colorectal adenomas, and whether CCND1 genotypes are associated with levels of epithelial cell proliferation in normal appearing colorectal mucosa.


   MATERIALS AND METHODS
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Study Design.
From 1994 to 1997, the MAP case-control study was conducted to assess the validity of colonic epithelial cell proliferation as a biomarker of risk for sporadic colorectal adenomas. Before beginning the study, MAP was approved by the Institutional Review Board of Wake Forest University School of Medicine in accordance with an assurance filed with and approved by the Department of Health and Human Services. Informed consent was obtained from each participant. Eligibility criteria for study subjects consisted of English-speaking adults from 30 to 74 years of age, of either sex, and any race who were scheduled for elective outpatient colonoscopy by one of three large gastroenterology practices in Winston-Salem and Charlotte, North Carolina. Patients were accrued over a 24-month period. Cases were identified as eligible colonoscopy patients who were determined to have study index pathologist-confirmed incident adenomatous polyps according to criteria adapted from the National Polyp Study (24) . Controls consisted of all eligible colonoscopy patients with no previous history of adenomas and who were found to be free of such tumors. People with familial polyposis, Gardner’s syndrome, ulcerative colitis, Crohn’s disease, bowel resection, newly diagnosed recurrent adenomatous polyps, and incident colon cancer were excluded as were patients with past or prevalent cancer other than nonmelanoma skin cancer.

Patients completed mailed questionnaires before their colonoscopies (to avoid bias) regarding family history of polyps or colon cancer, medical history, dietary information (via a semiquantitative Willett 153-item food frequency questionnaire), physical activity (via a modified Paffenbarger questionnaire), reproductive variables, body fat distribution, and their reason(s) for and the sequence of events leading to colonoscopy. Blood was drawn and stored at -70°C for possible later measurement of various genotypes.

Preparation for colonoscopy included a 12-h fast and bowel cleansing with polyethylene glycol (GoLYTELY). Subjects willing to undergo biopsies had four quadrant biopsies taken from normal appearing mucosa in the rectum (10 cm above the anus), sigmoid colon, and cecum for a total of 16 biopsies. Information recorded included number of polyps, polyp size, polyp type (adenoma, hyperplastic, mixed, or other), adenoma subtype (tubular, villous, or tubulovillous), and the degree of dysplasia. In addition, all normal tissue biopsies collected were analyzed for colonic epithelial cell proliferation using immunohistochemical detection of PCNA as previously described by Bostick et al. (25) .

Study Population and Sample Size.
Among all three clinical sites, 2246 colonoscopy patients were identified. Of these, 669 were eligible on initial screening (eligibility rate 29.8%), and of these, 633 were willing to discuss the study, 617 of these were contacted, and 417 of these signed consent and had study colonoscopies (consent rate 63.1%). Of the 417 participants, 259 had some type of polyp, and of these, 179 had adenomas. Nine of the 417 total patients were subsequently determined ineligible for the study, and an additional 8 patients had incident colon cancer and were not eligible for the primary case-control analyses; thus, 400 possible patients were available for genotypic analysis. Of these 400 patients, viable DNA was isolated from 374 (161 cases and 213 controls) for genotyping.

Laboratory Methods.
Genomic DNA was obtained from stored WBCs digested in 500 µl of lysis buffer [50 mM Tris/HCl (pH 8.5), 1 mM EDTA, 0.2% SDS, and 200 g/ml proteinase K] overnight at 55°C with shaking. The digestion was precipitated directly with isopropanol, and the pellets were washed with 70% ethanol. The genomic DNA pellets (50–100 µg) were dissolved in 300–800 µl of TE [10 mM Tris (pH 8)-1 mM EDTA] buffer, of which ~1 µl was used for each PCR reaction.

The CCND1 G/A polymorphism was detected by the PCR- RFLP method. According to the published sequence of the human CCND1 gene, we designed two primers (forward 5'-GTGAAGTTCATTTCCAATCCGC-3'; reverse 5'-GGGACATCACCCTCACTTAC-3'; Life Technologies, Inc.-Invitrogen) to amplify a 167-bp fragment of CCND1 gene at the junction of exon 4/intron 4. The PCR reactions were performed on a Perkin-Elmer GeneAmp System 9700 according to the manufacturer’s protocol. Specifically, these reactions were carried out in a 50-µl volume of 20 mM Tris-HCl (pH 8.4), 50 mM KCI, 1.0 mM MgCl2, 0.2 mM deoxynucleoside triphosphate, 1 unit of Taq polymerase (Life Technologies, Inc.-Invitrogen), and 0.4 µM of each oligonucleotide primer. The reactions were heated to 94°C for 1 min followed by 35 cycles of 94°C for 30 s, 60°C for 30 s, and 72°C for 40 s. At the end, the reactions were extended for 7 min at 72°C. For RFLP analyses, each PCR product was subjected to NciI digestion before electrophoresis. The DNA fragments were separated using 3% 2:1 Nusiev/SeaKem agarose gel. The allele types were determined as follows: a single 167-bp fragment for the AA genotype, two fragments of 145 and 22 bp for the GG genotype, and three fragments of 167, 145, and 22 bp for the AG genotype.

Statistical Analyses.
Allelic frequencies for polymorphic CCND1 A/G alleles were compared with those in previous study populations. Cyclin D1 genotype (GG, AG, AA) distributions for cases and controls were tested for adherence to the Hardy-Weinberg equilibrium.

All statistical inquiries were conducted using SAS Software version 8.2e from the SAS Institute in Cary, North Carolina. Descriptive comparisons (i.e., means, SD, frequencies as percents) of cases and controls were conducted using {chi}2 tests for categorical variables and analysis of covariance for continuous variables.

Logistic regression was used to calculate ORs and corresponding 95% CIs, adjusted only for age and sex, to estimate the strength of an association between CCND1 genotype and risk for incident sporadic colorectal adenomas. The effect of CCND1 genotype was analyzed using the a priori hypothesized low-risk, common GG genotype as the referent group. A test for trend was calculated across genotypes to detect a pattern of association.

Several risk factors were examined as possible confounders or effect modifiers of the CCND1 genotype-colorectal adenoma association. Among these were age, sex, race, body mass index, FHCC, smoking, alcohol consumption, NSAID use, total dietary intakes of fat, energy, calcium, sucrose, fiber, folate, meat, vegetables and fruit, and various antioxidant micronutrients. Criteria for inclusion of any covariate in the final model included: (a) biological plausibility; (b) whether it fit the model at P <= 0.1; and (c) whether it altered the OR for the primary exposure variable, CCND1 genotype, by 10% or more. Final models for genotype effects included only age and sex. Models involving assessing possible interactions between genotypes and various antiproliferative and proproliferative and other key risk factors included age, sex, FHCC, NSAIDs, smoking status, and total intakes of energy, calcium, and alcohol.

To examine separate and combined effects of CCND1 genotype and certain risk factors, stratified analyses were conducted. Continuous variables were dichotomized on median values for controls; furthermore, continuous dietary variables were categorized as sex specific. Criteria for assessing effect modifiers were based on previous literature, biological plausibility, and whether risk estimates differed substantially across strata.


   RESULTS
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
Selected characteristics of cases and controls are presented in Table 1Citation . On average, cases were more likely to be men, slightly older, and current smokers than controls. Controls were more likely to have histories of colon cancer in first-degree relatives and take NSAIDs or consume greater amounts of calcium.


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  		Table 1 Selected characteristics of cases and controls, MAP study, 1994–1997

 
Genotype analyses revealed equivalent allele distributions for cases (A = 42%, G = 58%) and controls (A = 38%, G = 62%). These frequencies were in accordance with those reported previously (12 , 13 , 19 , 22) . Hardy-Weinberg analyses detected no deviation from equilibrium among cases (P = 0.38) or controls (P = 0.78), and a {chi}2 test confirmed little difference between the groups (P = 0.19).

Age- and sex-adjusted associations of CCND1 genotypes and risk for colonic neoplasms are presented in Table 2Citation . A 50% risk increase (OR 1.5, 95% CI 1.0–2.4) was found to be associated with A allele carriers when compared with homozygous G individuals. This finding was unchanged with additional risk factor adjustment.


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  		Table 2 Age- and sex-adjusted associations of CCND1 genotypes and risk for incident sporadic colorectal adenomas, MAP study, 1994–1997

 
Age- and sex-adjusted associations of cyclin D1 genotypes and risk for incident sporadic adenomas according to adenoma characteristics are shown in Table 3Citation . The direct association with A allele carriers was stronger and statistically significant for multiple adenomas, and adenomas that were pedunculated, large, had higher levels of dysplasia, tubular, or located exclusively in the right colon.


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  		Table 3 Age- and sex-adjusted associations of CCND1 genotypes and risk for incident sporadic colorectal adenomas according to adenoma characteristics, MAP study, 1994–1997

 
Results of analyses to assess potential CCND1 genotype-environment interactions are shown in Table 4Citation . Interactions were suggested with age, sex, and alcohol consumption with statistically significant highest risk being found in A allele carriers who were also older (>57 years), men, or consumed more alcohol. These patterns were consistent with multiplicative interactions; however, the interaction terms were not statistically significant, perhaps because of the small sample size. Similarly, an additive interaction was found with smoking. Significant stronger inverse associations were found for NSAID use and calcium intake among those with the GG genotype.


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  		Table 4 Multivariate-adjusteda joint and combined associations of CCND1 genotypes and various risk factors for colorectal neoplasms and risk for incident sporadic colorectal adenomas, MAP study, 1994–1997

 
Intrarater scoring for colorectal epithelial cell proliferation was assessed by intraclass correlation coefficient and was 0.94. Mean levels of cellular proliferation did not differ according to genotype (data not shown). There was also no evidence that CCND1 genotype modified the association of cellular proliferation and risk for adenoma. Age of diagnosis of adenomas did not differ according to genotype (data not shown).


   DISCUSSION
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
The results of this study are consistent with a priori hypotheses of an increased risk for colorectal adenoma with the CCND1 variant allele. There was an overall 50% increase in risk for colorectal adenomas apparent for individuals with the A allele. Furthermore, the association was more pronounced and statistically significant among individuals with more advanced adenomas (i.e., >=1 cm, multiple, more dysplastic), suggesting that CCND1 may have not only a role in risk for adenomas but a strong role in tumor progression.

Our findings did not support an association between CCND1 genotype and colorectal epithelial cell proliferation in normal appearing colorectal mucosa. This suggests at least three possibilities: (a) our null finding may have been because of measurement error or chance; (b) there may be other mechanisms to explain the apparent association between CCND1 and risk for colorectal cancer; or (c) CCND1 genotype may have a minimal impact on epithelial cell proliferation in normal colorectal mucosa, but differences may be amplified in neoplastic tissue, thus promoting the progression from early to more advanced adenomas. On the one hand, our null finding is indirectly supported by the findings of no differences in cell proliferation by PCNA in 100 colorectal carcinomas according to genotype (23) ; but on the other hand, our findings of an adenoma-CCND1 genotype association were stronger for more advanced adenomas.

In addition, although our sample size was small and there were multiple comparisons, we found substantial, mostly statistically significant evidence that CCND1 genotype may modify associations of adenoma with age, sex, smoking, alcohol intake, NSAID use, and calcium intake. Among those with an A allele, risk was higher for men, smokers, alcohol users, and individuals > 57 years of age; and among those with the GG genotype, risk was much lower among those who took NSAIDs or consumed greater amounts of calcium.

Although one previous study of colon cancer in HNPCC carriers found that age of onset was associated with CCND1 genotype (21) , we found no such association among incident sporadic adenoma patients. This null finding, however, may be because of a lower correlation between age of adenoma onset and diagnosis in a general population than age of cancer incidence and diagnosis in HNPCC carriers.

The majority of tumor tissue studies conducted on cyclin D1 found overexpression in ~20–50% of adenomas and colorectal tumors (16 , 18 , 26, 27, 28) . A recent study found that cyclin D1 expression was increased in adenomas and additionally increased in malignant foci within adenomas (29) . These findings suggest that cyclin D1 overexpression may be an early-stage indicator in the multistage process of carcinogenesis (16 , 27) . Cyclin D1 overexpression was also found to be associated with such prognostic indicators as decreased survival and poor prognosis (23 , 30 , 31) . Still, all colorectal cancers do not overexpress cyclin D1, suggesting that the mechanism by which cyclin D1 is overexpressed may be limited to a subset of colorectal cancers, which is consistent with the heterogeneous pathways for developing colon cancer. However, alternatively, cyclin D1 overexpression may be lost during the progression from adenoma to carcinoma in some cases.

The mechanism for increased cyclin D1 expression in the colon is not through gene amplification as it is in parathyroid tumors (32) . CCND1 was found to be a polymorphic gene in which the normal occurring A870G single nucleotide mutation, located within the splice donor site in exon 4, is capable of influencing alternate splicing, ultimately leading to a truncated longer half-life protein (19) , and thus a sustained hyperproliferation that could promote carcinogenesis. However, as discussed above, our results suggest that this may be more evident in neoplastic, rather than normal colorectal tissue. Accumulation of the truncated cyclin D1 protein is primarily because of the absence of the PEST region responsible for rapid turnover via ubiquitin or calpain protease (33 , 34) .

The limited past literature on CCND1 gene polymorphism has been focused on its relevance to colorectal carcinoma. To our knowledge, there are no reports of investigations of CCND1 genotypes and risk for adenomas.

There have been five case-control studies that have investigated CCND1 genotypes and risk for colorectal carcinoma. Of these, two found statistically significant associations of CCND1 genotype and risk for colorectal cancer. The first, a hospital-based study of 156 incident sporadic colorectal cancer patients and 152 controls, reported a positive association with the A allele and risk for colorectal cancer (21) ; however, the association was only significant for the homozygous AA genotype, suggesting a recessive model of inheritance rather than the dominant A allele model suggested by our study and that of Betticher et al. (19) . The second, a genetic center study, involved both sporadic (n = 128) and familial (n = 206) colorectal cancer cases and 171 cancer-free controls (22) . The only statistically significant positive association was that between the dominant A allele and risk for familial colorectal cancer, a finding that was stronger in non-HNPCC cases than in HNPCC cases. Two other case-control studies, an 86-person (49 cases, 37 controls) study of HNPCC family members who were mutation carriers, and a 332-participant (146 cases, 186 controls) Finnish HNPCC family mutation carrier-based study, found significant associations among A allele carriers (12) and cyclin D1 transcripts (13) with age of colorectal cancer onset for HNPCC mutation carriers, respectfully. The fifth, a case-control study of CCND1 and colorectal cancer (n = 201) conducted on 100 cases identified from a tumor bank and 101 controls selected from blood donors, found a positive association with decreased survival time and increased protein expression but not genotype (23) . These findings suggest that CCND1 may influence risk for sporadic, familial, and HNPCCs.

This study has several strengths. It is, to our knowledge, the first to investigate the association of the novel polymorphic gene, CCND1, and colorectal adenomas. We were also able to assess this association according to adenomas along the spectrum of malignant potential and in interaction with various anti proliferative and proproliferative and other key risk factors. Other strengths of the study included minimizing recall bias by administration of questionnaires before diagnosis and avoiding case/control misclassification by full colonoscopic examination of all patients and index pathologist confirmation of all cases.

This study also has several limitations. First, the parent study was clinic based, and as a result, the study population may not be representative of the general population and may be at higher risk for colorectal neoplasms. Therefore, it is not unlikely that some patients receiving colonoscopies, who were classified as controls, will later develop polyps and thus had risk similar to that of the cases. Also, not uncommon in this type of case-control study, family history of colorectal cancer in first-degree relatives was highest among controls, thus possibly introducing a family history bias. However, these limitations would tend to attenuate associations; therefore, our findings may actually underestimate the influence of CCND1 genotypes on risk for colorectal neoplasms. Second, our sample size was small, and we made multiple, albeit a priori biologically plausible, comparisons. However, the findings were generally statistically significant and consistent with a priori hypotheses and thus warrant further study. Third, using genotype to assess risk may result in attenuated associations because genotypes do not always correspond to the expressed phenotypes, as suggested by the lack of association between CCND1 genotype and colorectal epithelial cell proliferation in normal appearing mucosa. Such validations are needed to establish whether risk estimates can reasonably be determined by genotype alone.

In summary, this study of CCND1 genotypes and risk for colorectal adenomas—the first to our knowledge—in conjunction with previous, albeit small, case-control studies of CCND1 and colon cancer, support hypotheses that the CCND1 A allele is associated with increased risk for colorectal neoplasms. Our findings also suggest that the association of this allele with adenomas is strongest for more advanced adenomas, possibly indicating that the role for the A allele may be stronger for adenoma progression than initiation. Our findings were not supportive of the hypothesis that differential risk for adenoma according to genotype is explained by differences in colorectal epithelial cell proliferation in normal appearing colorectal mucosa. Finally, our findings also suggest that CCND1 genotypes may differentially influence risk from various environmental risk factors.


   FOOTNOTES
 
Grant support: National Cancer Institute Grant R01 CA66539 (to R. M. B.).

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.

Requests for reprints: Roberd M. Bostick, Division of Population Studies, 15 Medical Park, Suite 301, Columbia, SC 29203. Phone: (803) 434-1613; Fax: (803) 434-1626; E-mail: rbostick{at}gw.mp.sc.edu

7 The abbreviations used are: TCF/LEF, T-cell factor/lymphoid enhancing factor; NSAID, nonsteroidal anti-inflammatory drug; APC, adenomatous polyposis coli; MMR, mismatch repair; MAP, Markers for Adenomatous Polyps; HNPCC, hereditary nonpolyposis colorectal cancer; FHCC, family history of colon cancer; PCNA, proliferating cell nuclear antigen; OR, odds ratio; CI, confidence interval. Back

Received 7/ 3/03. Revised 9/ 2/03. Accepted 9/ 4/03.


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Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 

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