OROncology Reports1021-335X1791-2431D.A. Spandidos10.3892/or.2013.2926or-31-02-0995ArticlesPolymorphisms of the pri-miR-34b/c promoter and TP53 codon 72 are associated with risk of colorectal cancerOHJISU1*KIMJONG WOO2*LEEBO EUN3JANGMOON JU1CHONGSO YOUNG1PARKPIL WON1HWANGSEONG GYU13OHDOYEUN13KIMNAM KEUN3
Department of Internal Medicine, CHA University, Seongnam, Republic of Korea
Department of Surgery, CHA University, Seongnam, Republic of Korea
Institute for Clinical Research, CHA Bundang Medical Center, CHA University, Seongnam, Republic of KoreaCorrespondence to: Professor Nam Keun Kim, Institute for Clinical Research, CHA Bundang Medical Center, CHA University, 59 Yatap-ro, Bundang-gu, Seongnam 463-712, Republic of Korea, E-mail: nkkim@cha.ac.kr
The microRNA (miR)-34 family is a direct transcriptional target of tumor-suppressor TP53 and loss of miR-34 function may impair TP53-mediated cell cycle arrest and apoptosis. In the present study, we investigated whether the single nucleotide polymorphisms (SNPs) rs4938723 (T>C) in the promoter region of miR-34b/c and Arg72Pro (G>C) in codon 72 of TP53 are independently or complementarily associated with the risks and clinical outcomes of colorectal cancer (CRC) and whether the combined effect of these SNPs and metabolic risk factors are related to CRC. We evaluated the SNPs in 545 CRC patients and 428 healthy controls using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequence analysis. We found that the GC and GC/CC genotypes of TP53 Arg72Pro were associated with decreased risk of CRC (adjusted OR = 0.727 for GC; OR = 0.735 for GC/CC). The combined genotypes of TT-GC and CC-GG were significantly associated with reduced CRC risk (adjusted OR = 0.628 for TT-GC; OR = 0.381 for CC-GG, respectively). The SNP rs4938723 and diabetes mellitus (DM) together were associated with an increased CRC risk, but the SNP TP53 Arg72Pro CC with DM showed a protective effect against CRC. These findings indicate that rs4938723 in the promoter region of pri-miR-34b/c and the SNP in TP53 codon 72 were related to decreased risk of CRC in the population studied and those metabolic diseases and genetic variants influence each other with regard to CRC susceptibility.
microRNA-34b/cTP53 Arg72Propolymorphismrisk and prognosismetabolic diseasescolorectal cancerIntroduction
Colorectal cancer (CRC) is one of the most common types of cancer (the third and fourth most common cancer in women and men, respectively) in the world and the second leading cause of cancer-related mortality in developed countries. More than 1.2 million cases are diagnosed globally each year, with ~600,000 deaths (1,2). In order to find new diagnostic and therapeutic tools to reduce CRC-related morbidity, it is key to understand the etiology and biology of CRC. Progression to CRC is caused by an accumulation of various genetic and epigenetic alterations, leading to transformation from a normal tissue to a malignant, and potentially metastatic, tumor. CRC develops through two main genetic pathways that are characterized by different forms of genomic instability, the chromosomal instability (CIN, 85%) and microsatellite instability (MSI, 15%) pathways (3–5). Many tumor-suppressor genes and oncogenes have been described, and the discovery of new tumor markers, including those for CRC, continues at a rapid pace.
A new group of biomarkers, microRNAs (miRs), has recently been established. miRs are 20–25-nucleotide non-coding RNAs that negatively regulate gene transcription at the transcriptional or post-transcriptional level by interacting with the 3′ untranslated regions (UTRs) of specific messenger RNAs (mRNAs) and are key modulators in the control of biological processes, such as cell development, differentiation, proliferation and apoptosis (6–9). Estimates based on bioinformatics and microarray analyses suggest that >30% of all genes are regulated by multiple miRs (10). miR-34s form an evolutionarily conserved miR family, with miR-34a, miR-34b and miR-34c occurring in vertebrates. The miR-34a and miR-34b/c loci are regulated directly by interaction with TP53, which induces apoptosis, cell cycle arrest and senescence (11–13). In addition, reduced miR-34a expression is a frequent feature of both pancreatic tumors and neuroblastomas (14,15) and reduced miR-34b/c expression has been observed in non-small cell lung cancer (16). CpG methylation of miR-34b/c has been found in CRC (2,17), oral squamous cell carcinoma (18), and malignant melanoma, where it correlated with metastatic potential (19).
The downregulation of miR-34 family members in cancer suggests that these miRs function as tumor-suppressor genes, suggesting a possible role as prognostic markers (20). Several mechanisms regulating miR expression, including gene amplification, deletion, epigenetic alterations and single-nucleotide substitution, have been implicated, but not demonstrated (20,21). Although single nucleotide polymorphisms (SNPs) in miRs are not considered functionally important, nucleotide variations in primary (pri)- or precursor (pre)-miRs may affect miR processing and modify miR expression (22). Recently, studies reported that a potentially functional SNP, rs4938723, in the promoter region of pri-miR-34b/c may contribute to susceptibility to hepatocellular carcinoma (23), CRC (24), endometrial cancer (25) and survival in breast cancer (26). However, there are few reports on the relationship between SNPs in the miR-34b/c promoter and risk and prognostic significance in CRC patients.
In the present study, we investigated whether the SNPs rs4938723 (T>C) in the promoter region of miR-34b/c and Arg72Pro (G>C) in codon 72 of TP53 are independently or complementarily associated with the risk and clinical outcomes of CRC and whether the combined effect of these SNPs and metabolic risks (diabetes and hypertension) is related to progression of CRC, which is known to be associated with metabolic disease (27,28) in the Korean population.
Materials and methodsPatients and clinical samples
From June 2000 to January 2009, blood samples were collected from 545 patients diagnosed with CRC at CHA Bundang Medical Center of CHA University in South Korea. We retrospectively obtained information concerning the age; gender; underlying conditions [hypertension (HTN), diabetes mellitus (DM), body mass index (BMI), smoking and alcohol consumption]; tumor size, stage and site; time to progression; and time to mortality. We estimated homocysteine and folic acid levels. The American Joint Committee on Cancer: Classification and Stage Groupings, 7th edition was used for tumor assessment. The cancer-free control group consisted of 428 individuals who were randomly selected from participants in a health-screening program to exclude those with a history of cancer and other medical diseases. All study subjects provided written consent and all were ethnic Koreans. The subjects’ recruitment was approved by the Institutional Review Board of CHA Bundang Medical Center.
Genotyping
DNA was extracted from leukocytes using a G-DEX™ II Genomic DNA Extraction kit (iNtRON Biotechnology, Seongnam, Korea), according to the manufacturer’s instructions. The SNPs miR-34b/c rs4938723 and TP53 Arg72Pro rs1042522 were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assays. Primer sequences used for amplification of rs4938723 were: (forward) 5′-CCT CTG GGA ACC TTC TTT GAC CAA T-3′ and (reverse) 5′-TGA GAT CAA GGC CAT ACC ATT CAA GA-3′. Primer sequences used for amplification of TP53 Arg72Pro were: (forward) 5′-TTG CCG TCC CAA GCA ATG GAT GA-3′ and (reverse) 5′-TCT GGG AAG GGA CAG AAG ATG AC-3′. For each of the polymorphisms, 30% of the PCR assays were randomly chosen and repeated, followed by DNA sequencing to validate the RFLP findings. Sequencing was performed using an ABI 3730xl DNA Analyzer (Applied Biosystems, Foster City, CA, USA). The concordance of the quality control samples was 100%.
Statistical analysis
The genotypes for each SNP were analyzed as a three-group categorical variable (reference model) and were also grouped according to the dominant and recessive model. Odds ratios (ORs), adjusted odds ratios (AORs) and 95% confidence intervals (CIs) were used to calculate the strength of association. To analyze baseline characteristics, we used a Chi-square test for categorical data when comparing patient and control baseline data. Multivariate analysis was performed to select independent risk factors for CRC among genotypes and clinical variables using logistic regression analysis. The overall survival was compared using the Kaplan-Meier method and potential variables were verified by multivariate analysis using a Cox regression model. All tests were two-tailed, and a P-value <0.05 was deemed to indicate a statistically significant difference. Analyses were performed using GraphPad Prism 4.0 (GraphPad Software, San Diego, CA, USA) and MedCalc version 11.1.1.0 (MedCalc Software, Ostend, Belgium). The distribution of allele frequencies for the rs4938723 (T>C) and TP53 Arg72Pro (G>C) gene polymorphisms were calculated by Chi-square test to determine whether the observed genotype distributions conformed to the expected Hardy-Weinberg equilibrium (29).
ResultsPopulation
Baseline characteristics of patients with CRC and controls are shown in Table I. The mean age was 62 years in CRC patients and 61 years in controls. Of the CRC patients, 302 (55.4%) were male. Approximately 61.5% of CRC cases had HTN and 33.6% had DM; these were significantly higher values than observed among controls (34.1 and 6.1%, respectively) (P<0.001). However, the number of smokers in the control group exceeded that for CRC patients. Tumors occurred most frequently in the rectum and proximal colon. With regard to tumor-node-metastasis (TNM) staging, the majority of patients (80.7%) had stage II–III disease.
Variant genotypes of TP53 codon 72 are related to reduced CRC risk
Genotype frequencies of miR-34b/c rs4938723 and TP53 Arg72Pro rs1042522 in CRC patients and controls are shown in Table II. In multivariate analysis, the variant genotypes of TP53 Arg72Pro GC and GC/CC were associated with a significantly decreased risk of CRC compared with the wild-type GG genotype (AOR = 0.727, 95% CI = 0.550–0.960 for GC; AOR = 0.735, 95% CI = 0.565–0.958 for GC/CC). However, no overall association was observed between miR-34b/c rs4938723 and CRC risk in our study population. The observed genotype frequencies for miR-34b/c rs4938723 (T>C) and TP53 Arg72Pro (G>C) polymorphisms in the cases and controls were all as expected for Hardy-Weinberg equilibrium (P>0.05).
Combined genotype effects of rs4938723 and TP53 (TT/GC and CC/GG) are associated with decreased CRC risk in colon cancer patients
Combined genotype analyses were conducted to evaluate the combined effects of the two polymorphisms on the risk of CRC (Table III). Nine combined genotypes were estimated from the two polymorphisms in CRC patients. In the multivariate analysis, combined genotypes TT/GC and CC/GG were associated with significantly decreased CRC risk when compared with the wild-type TT/GG genotype (AOR = 0.628, 95% CI = 0.422–0.934 for TT/GC; AOR = 0.381, 95% CI = 0.183–0.793 for CC/GG, respectively). This association was observed only in patients with colon, not rectal, cancer and, moreover, was observed only in proximal colon cancer patients (data not shown).
SNP rs4938723 with DM is associated with increased CRC risk
Table IV shows CRC risk by combined genetic-environmental effects (HTN, DM, homocysteine and folic acid). TP53 Arg72Pro GG and all genotypes of rs4938723 with HTN were significantly associated with increased risk of CRC. All DM patients, in particular, showed strong positive association with CRC, regardless of genotype. The polymorphism rs4938723 with DM was associated with a significantly increased CRC risk compared with wild-type TT and TP53 Arg72Pro CC with DM significantly decreased the risk when compared with wild-type GG. No differences were observed between homocysteine and folic acid levels for any genotype (data not shown). A homocysteine level >11.7 μmol/l was associated with increased risk of CRC in rs4938723 TC and TC/CC, but with decreased risk for TP53 Arg72Pro SNPs. Folic acid level <4.45 ng/ml was associated with an increased CRC risk, but the rs4938723 and TP53 Arg72Pro polymorphisms decreased the risk of CRC when compared with wild-type (Table IV).
Polymorphisms rs4938723 and TP53 Arg72Pro show a trend toward, but are not significantly associated with, improved survival
The 3-year survival rate was estimated in each miR-34b/c rs4938723 and TP53 Arg72Pro patient group (Table V). In the multivariate analysis, overall survival of the variant miR-34b/c rs4938723 and TP53 Arg72Pro genotypes was more evident in subjects carrying polymorphisms than in wild-type. However, we did not find a significant association between miR-34b/c rs4938723 and TP53 Arg72Pro genotypes and survival of CRC (Fig. 1). Stratified analyses (Table VI) showed that the risk reduction effect of the variant TP53 Arg72Pro GC/CC genotypes was significant in subjects without HTN (AOR = 0.635, 95% CI = 0.418–0.966). However, no significant interactions were observed between miR-34b/c rs4938723 TC/CC and other clinical features.
Discussion
In the present study on CRC in a Korean population, we investigated the correlation between the SNPs rs4938723, in the promoter region of pri-miR-34b/c and TP53 Arg72Pro and the risk of CRC. We found that the TP53 Arg72Pro polymorphism was significantly associated with a decreased risk of CRC. The combined genotypes rs4938723 CC and TP53 GG had a tendency to protect individuals against CRC. This finding suggests that SNPs in the promoter regions of miRs may have important roles in the etiology of CRC, providing novel biomarkers for malignancies. This tendency was shown for cancer of the colon, particularly the proximal colon, but not for rectal cancer. Further investigation of the relationship between the site of cancer and genotype is required. Recently, studies reported that TP53 influences the expression of miR-34b/c in a one-way relationship. However, rs4938723 TT and TP53 Arg72Pro GC were also associated with decreased CRC risk in the present study, which suggests that TP53 and miR-34b/c may interact bidirectional and supports the concept that CRC is a complex disease involving multiple genes.
We also found that HTN or DM was associated with a significantly increased risk of CRC, regardless of genotype (Table IV). SNP rs4938723 with DM was associated with increased CRC risk when compared with wild-type, but TP53 Arg72Pro CC with DM was associated with decreased risk when compared with wild-type. Moreover, decreased levels of homocysteine and folic acid showed a positive correlation with the risk of CRC (Table VII). At folic acid levels of <4.45 ng/ml, both the rs4938723 and TP53 codon 72 SNPs were related to a decreased risk of CRC. At homocysteine levels >11.70 μmol/l, rs4938723 was positively correlated with CRC risk, but the TP53 codon 72 SNP was inversely correlated. These findings suggest that genetic factors and environmental factors, such as metabolic diseases, influence each other in CRC and may implicate miRs as pathophysiologic linkers/modulators between metabolic syndrome and cancer.
Rs4938723 is located within the CpG island of pri-miR-34b/c (423-bp upstream of the transcription start site), making it a potential binding site for GATA-X transcription factors. According to the web-based SNP analysis tool TFSEARCH 1.3, GATA family members bind to promoters of many genes and directly activate or suppress expression of target genes and may be involved in carcinogenesis (30). Therefore, SNP rs4938723 (T>C) may affect the expression level of miR-34b/c. Ectopic miR-34b/c caused cell cycle arrest in the G1 phase (31) and miR-34b/c inhibited proliferation and colony formation in soft agar (11). The relationship between miR expression in tumors and prognosis, with regard to both the treatment response rate and survival, is of increasing interest. This polymorphism has been reported to be associated with susceptibility to hepatocellular carcinoma (23), CRC (24), survival of breast cancer (26), renal cell carcinoma (32), non-small cell lung (33) and oral cancer (18).
Arg72Pro is the most widely investigated of the variations in the TP53 gene. The 72Arg allele induces apoptosis more efficiently than the 72Pro allele (34). It has been reported that homozygosity for Pro of TP53 Arg72Pro is potentially a risk factor for cancer of the lung, esophagus, stomach, breast, nasopharynx, urothelium and prostate (35–37). In CRC, meta-analysis was performed to estimate the effect of the TP53 Arg72Pro polymorphism and CRC risk, and no significant association was identified (38). In contrast to that result, we found a negative relationship between the TP53 Arg72Pro polymorphism and CRC risk in the Korean population.
Our study has several limitations. First, the enrolled patients were selected at a single institution in Korea and the sample size may limit the statistical power, especially for evaluating the connection between SNPs and CRC risk and survival in detailed genotype subgroups. Second, as the study was hospital-based, the cases and controls could not be representative of the general population. This possible selection bias could not be avoided. Finally, the mechanisms underlying the effects of genetic polymorphisms on the levels of pre/mature miRs and the identity of the miR target genes remain unknown and should be determined in future studies.
In conclusion, although we identified no significant prognostic value of SNPs for CRC, we found that SNPs rs4938723 and TP53 Arg72Pro show a trend toward improved survival and that the TP53 Arg72Pro CC genotype and dominant model (GC + CC) significantly decrease the risk of CRC. Additionally, the combined genotype rs4938723 and TP53 Arg72Pro (TT/GC and CC/GG) was significantly associated with decreased CRC risk in the Korean population. Also, SNP rs4938723 with DM was more closely related to increased CRC risk than the wild-type genotype. Future studies in multiethnic populations are warranted to validate our results and to define the functional effects of these SNPs on CRC.
Acknowledgements
This study was partially supported by a National Research Foundation (NRF) of Korea Grant funded by the Korean Government (2009-0075784 and 2012R1A1A2007033), and a Priority Research Centers Program Grant, administered by the NRF and funded by the Ministry of Education (2009-0093821), Republic of Korea.
ReferencesSiegelRNaishadhamDJemalACancer statistics, 2012CA Cancer J Cin6210292012HrašovecSGlavačDMicroRNAs as novel biomarkers in colorectal cancerFront Genet31802012CunninghamJMChristensenERTesterDJHypermethylation of the hMLH1 promoter in colon cancer with microsatellite instabilityCancer Res58345534601998ToyotaMAhujaNOhe-ToyotaMHermanJGBaylinSBIssaJPCpG island methylator phenotype in colorectal cancerProc Natl Acad Sci USA96868186861999ShenLToyotaMKondoYIntegrated genetic and epigenetic analysis identifies three different subclasses of colon cancerProc Natl Acad Sci USA10418654186592007IorioMVCroceCMMicroRNA dysregulation in cancer: diagnostics, monitoring and therapeutics. A comprehensive reviewEMBO Mol Med41431592012FaziFNerviCMicroRNA: basic mechanisms and transcriptional regulatory networks for cell fate determinationCardiovasc Res795535612008MeisterGTuschlTMechanisms of gene silencing by double-stranded RNANature4313433492004PillaiRSBhattacharyyaSNFilipowiczWRepression of protein synthesis by miRNAs: how many mechanisms?Trends Cell Biol171181262007LimLPLauNCGarrett-EngelePMicroarray analysis shows that some microRNAs downregulate large numbers of target mRNAsNature4337697732005CorneyDCFlesken-NikitinAGodwinAKWangWNikitinAYMicroRNA-34b and MicroRNA-34c are targets of p53 and cooperate in control of cell proliferation and adhesion-independent growthCancer Res67843384382007HeLHeXLimLPA microRNA component of the p53 tumour suppressor networkNature447113011342007TarasovVJungPVerdoodtBDifferential regulation of microRNAs by p53 revealed by massively parallel sequencing: miR-34a is a p53 target that induces apoptosis and G1-arrestCell Cycle6158615932007ChangTCWentzelEAKentOATransactivation of miR-34a by p53 broadly influences gene expression and promotes apoptosisMol Cell267457522007WelchCChenYStallingsRLMicroRNA-34a functions as a potential tumor suppressor by inducing apoptosis in neuroblastoma cellsOncogene26501750222007BommerGTGerinIFengYp53-mediated activation of miRNA34 candidate tumor-suppressor genesCurr Biol17129813072007ToyotaMSuzukiHSasakiYMaruyamaRImaiKShinomuraYTokinoTEpigenetic silencing of microRNA-34b/c and B-cell translocation gene 4 is associated with CpG island methylation in colorectal cancerCancer Res68412341322008KozakiKImotoIMogiSOmuraKInazawaJExploration of tumor-suppressive microRNAs silenced by DNA hypermethylation in oral cancerCancer Res68209421052008LujambioACalinGAVillanuevaAA microRNA DNA methylation signature for human cancer metastasisProc Natl Acad Sci USA10513556135612008KongYWFerland-McColloughDJacksonTJBushellMmicroRNAs in cancer managementLancet Ooncol13e249e2582012LujambioARoperoSBallestarEGenetic unmasking of an epigenetically silenced microRNA in human cancer cellsCancer Res67142414292007IwaiNNarabaHPolymorphisms in human pre-miRNAsBiochem Biophys Res Commun331143914442005XuYLiuLLiuJA potentially functional polymorphism in the promoter region of miR-34b/c is associated with an increased risk for primary hepatocellular carcinomaInt J Cancer1284124172011GaoLBLiLJPanXMLiZHLiangWBBaiPZhuYHZhangLA genetic variant in the promoter region of miR-34b/c is associated with a reduced risk of colorectal cancerBiol Chem3944154202013HirokiESuzukiFAkahiraJNagaseSItoKSugawaraJMikiYSuzukiTSasanoHYaegashiNMicroRNA-34b functions as a potential tumor suppressor in endometrial serous adenocarcinomaInt J Cancer131E395E4042012BensenJTTseCKNyanteSJBarnholtz-SloanJSColeSRMillikanRCAssociation of germline microRNA SNPs in pre-miRNA flanking region and breast cancer risk and survival: the Carolina Breast Cancer StudyCancer Causes Control24109911092013AhmedRLSchmitzKHAndersonKERosamondWDFolsomARThe metabolic syndrome and risk of incident colorectal cancerCancer10728362006MoghaddamAAWoodwardMHuxleyRObesity and risk of colorectal cancer: a meta-analysis of 31 studies with 70,000 eventsCancer Epidemiol Biomarkers Prev16253325472007Wittke-ThompsonJKPluzhnikovACoxNJRational inferences about departures from Hardy-Weinberg equilibriumAm J Hum Genet769679862005ChouJProvotSWerbZGATA3 in development and cancer differentiation: cells GATA have it!J Cell Physiol22242492010XiangYFanSCaoJHuangSZhangLPAssociation of the microRNA-499 variants with susceptibility to hepatocellular carcinoma in a Chinese populationMol Biol Rep39701970232012LinJHorikawaYTamboliPClagueJWoodCGWuXGenetic variations in microRNA-related genes are associated with survival and recurrence in patients with renal cell carcinomaCarcinogenesis31180518122010HuZChenJTianTGenetic variants of miRNA sequences and non-small cell lung cancer survivalJ Clin Invest118260026082008DumontPLeuJIDella PietraACIIIGeorgeDLMurphyMThe codon 72 polymorphic variants of p53 have markedly different apoptotic potentialNat Genet333573652003IrarrázabalCERojasCAracenaRMárquezCGilLChilean pilot study on the risk of lung cancer associated with codon 72 polymorphism in the gene of protein p53Toxicol Lett14469762003KurodaYTsukinoHNakaoHImaiHKatohTp53 codon 72 polymorphism and urothelial cancer riskCancer Lett18977832003SuzukiKMatsuiHOhtakeNA p53 codon 72 polymorphism associated with prostate cancer development and progression in JapaneseJ Biomed Sci104304352003DahabrehIJLinardouHBouzikaPVarvarigouVMurraySTP53 Arg72Pro polymorphism and colorectal cancer risk: a systematic review and meta-analysisCancer Epidemiol Biomarkers Prev19184018472010
(A) Overall survival, (B) relapse free survival according to miR-34b/c rs4938723 polymorphisms in CRC patients. CRC, colorectal cancer.
Baseline characteristics of CRC patients and controls.
Variable n (%)
Controls (n=428)
CRC patients (n=545)
OR (95% CI)
P-valuea
Age (years, mean ± SD)
60.75±11.73
62.07±12.15
1.008 (0.998–1.019)
0.134
Gender (male)
172 (40.2)
302 (55.4)
1.379 (1.100–1.729)
0.005
Hypertension
146 (34.1)
335 (61.5)
1.802 (1.429–2.272)
<0.0001
Diabetes mellitus
26 (6.1)
183 (33.6)
10.120 (6.657–15.380)
<0.0001
Body mass index ≥25.0 kg/m2
96 (22.4)
143 (26.2)
1.170 (0.877–1.560)
0.285
Smoking
140 (32.7)
125 (22.9)
0.701 (0.534–0.921)
0.010
Tumor size ≥5 cm
NA
319 (58.5)
Tumor site
Proximal colon
NA
180 (33.0)
Distal colon
NA
123 (22.6))
Mixed colon
NA
7 (1.3)
Rectum
NA
225 (41.3)
TNM stage
I
NA
55 (10.1)
II
NA
228 (41.8)
III
NA
212 (38.9)
IV
NA
49 (9.0)
Chi-square test for categorical data, two-sided t-test for continuous data.
CRC, colorectal cancer; OR, odds ratio; CI, confidence interval; SD, standard deviation; TNM, tumor-node-metastasis; NA, not applicable.
Genotype frequencies of miR-34b/c rs4938723 and TP53 Arg72Pro in CRC patients and controls.
Characteristics n (%)
Controls (n=428)
CRC patients (n=545)
AOR (95% CI)a
P-valueb
P-valuec
Colon (n=310)
AOR (95% CI)a
P-valueb
Rectum (n=230)
AOR (95% CI)a
P-valueb
miR-34b/c rs4938723
TT
216 (50.5)
272 (49.9)
1.000
158 (51.0)
1.000
114 (49.6)
1.000
TC
171 (40.0)
233 (42.8)
1.083 (0.830–1.414)
0.557
0.557
131 (42.3)
1.045 (0.769–1.420)
0.777
98 (42.6)
1.101 (0.785–1.542)
0.578
CC
41 (9.5)
40 (7.3)
0.780 (0.487–1.250)
0.302
0.302
21 (6.7)
0.700 (0.398–1.232)
0.216
18 (7.8)
0.859 (0.471–1.569)
0.621
Dominant model (TT vs. TC + CC)
1.025 (0.796–1.321)
0.848
0.848
0.978 (0.730–1.311)
0.883
1.052 (0.763–1.451)
0.758
Recessive model (TT + TC vs. CC)
0.754 (0.478–1.190)
0.225
0.450
0.687 (0.397–1.188)
0.179
0.821 (0.459–1.467)
0.505
HWE P
0.402
0.301
0.376
0.628
TP53 Arg72Pro
GG
145 (33.9)
222 (40.7)
1.000
130 (41.9)
1.000
88 (38.3)
1.000
GC
218 (50.9)
247 (45.3)
0.727 (0.550–0.960)
0.025
0.050
135 (43.5)
0.675 (0.489–0.931)
0.016
111 (48.3)
0.827 (0.582–1.175)
0.289
CC
65 (15.2)
76 (14.0)
0.757 (0.511–1.120)
0.164
0.302
45 (14.6)
0.769 (0.491–1.204)
0.251
31 (13.4)
0.762 (0.459–1.265)
0.294
Dominant model (GG vs. GC + CC)
0.735 (0.565–0.958)
0.023
0.046
0.699 (0.516–0.946)
0.020
0.815 (0.583–1.138)
0.229
Recessive model (GG + GC vs. CC)
0.903 (0.631–1.293)
0.577
0.577
0.947 (0.627–1.431)
0.797
0.860 (0.542–1.366)
0.523
HWE P
0.250
0.583
0.305
0.667
Adjusted by age, gender, hypertension and diabetes mellitus.