Prognostic Impact of Circulating Tumor Cell Detected Using a Novel Fluidic Cell Microarray Chip System in Patients with Breast Cancer.

Various types of circulating tumor cell (CTC) detection systems have recently been developed that show a high CTC detection rate. However, it is a big challenge to find a system that can provide better prognostic value than CellSearch in head-to-head comparison. We have developed a novel semi-automated CTC enumeration system (fluidic cell microarray chip system, FCMC) that captures CTC independently of tumor-specific markers or physical properties. Here, we compared the CTC detection sensitivity and the prognostic value of FCMC with CellSearch in breast cancer patients. FCMC was validated in preclinical studies using spike-in samples and in blood samples from 20 healthy donors and 22 breast cancer patients in this study. Using spike-in samples, a statistically higher detection rate (p=0.010) of MDA-MB-231 cells and an equivalent detection rate (p=0.497) of MCF-7 cells were obtained with FCMC in comparison with CellSearch. The number of CTC detected in samples from patients that was above a threshold value as determined from healthy donors was evaluated. The CTC number detected using FCMC was significantly higher than that using CellSearch (p=0.00037). CTC numbers obtained using either FCMC or CellSearch had prognostic value, as assessed by progression free survival. The hazard ratio between CTC+ and CTC- was 4.229 in CellSearch (95% CI, 1.31 to 13.66; p=0.01591); in contrast, it was 11.31 in FCMC (95% CI, 2.245 to 57.0; p=0.000244). CTC detected using FCMC, like the CTC detected using CellSearch, have the potential to be a strong prognostic factor for cancer patients.

Pharmacogenetics of hormone replacement therapy for climacteric symptoms.

Hormone replacement therapy (HRT) is highly effective for women suffering from climacteric symptoms, with occasionally severe side effects. To determine which women needs HRT for climacteric symptoms indeed, pharmacogenetical approach for HRT was performed. Under the condition of minimal HRT, 33 patients required HRT for more than 1 year and the remaining 156 did not. Three single nucleotide polymorphisms (SNPs) in estrogen receptor alpha (ERalpha) gene and 3 SNPs and a microsatellite polymorphism in estrogen receptor beta (ERbeta) gene were analyzed using LightTyper and PCR. Homozygous for 18 CA repeats of D14S1026 (OR 8.00, 95% CI 2.56-25.02, P<0.001) and rs1256049 (OR 6.35, 95% CI 2.38-16.92, P=0.004) in ERbeta associated with minimal HRT. In contrast, rs1271572 in 789bp upstream region of ERbeta (OR 0.30, 95% CI 0.14-0.65, P=0.002) gene decreased HRT. rs2228480 in ERalpha gene also increased HRT. Tailored decisions can be expected on the future use of HRT referring genetic polymorphisms of individuals.

Influence of CYP11B2 gene polymorphism on the prevalence of hypertension and the blood pressure in Japanese men: interaction with dietary salt intake.

AIMS: CYP11B2 gene encodes a key enzyme for the production of aldosterone. Our aim is to investigate the association of -344T/C polymorphism with hypertension in Japanese men. The interaction between genotypes and dietary salt intake was also considered. METHODS: Three hundred and ten Japanese male workers participated in this study. Daily salt intake was calculated from a food frequency questionnaire. Melting curve analysis was used to determine CYP11B2 genotypes. RESULTS: There was a significant association between the CT + TT genotype and higher prevalence of hypertension (odds ratio: 3.03; p = 0.014). The association presented in a recessive manner and was strongest in the high-salt intake group (odds ratio: 9.44; p = 0.049). Only in the high-intake group, systolic blood pressure (SBP) was significantly higher in the CT + TT group than in the CC group (p = 0.038). The SBP had a positive correlation with salt intake in the CT + TT group (p for linear trend = 0.021), but not in the CC group (p for interaction = 0.011). CONCLUSIONS: CYP11B2 gene -344C/T polymorphism affects the risk of hypertension in Japanese men and high-salt intake levels strengthen this association. This gene-diet interaction warrants further study to elucidate the efficacy of salt restriction as an antihypertensive therapy in different genotypes.

The interaction between the interleukin 6 receptor gene genotype and dietary energy intake on abdominal obesity in Japanese men.

Previous reports have shown that the Asp358Ala (T/G) polymorphism of the interleukin 6 receptor (IL6R) gene is associated with obesity and type 2 diabetes mellitus, but few studies have examined this association in the Japanese population. We performed the current study to investigate the relationship between the IL6R Asp358Ala (T/G) polymorphism and obesity in healthy Japanese men. Two hundred eighty-five healthy Japanese men (age, 46.1 +/- 11.5 years [mean +/- SD]; waist circumference [WC], 83.9 +/- 9.3 cm; body mass index, 23.3 +/- 3.3 kg/m(2)) employed by a Japanese company were enrolled in this study. Height, weight, and WC were measured, and daily energy intake levels were assessed by self-reported questionnaires. Genotyping of polymorphisms was performed by using melting curve analysis; no association was found between IL6R genotype and WC or body mass index. However, when the subjects were stratified by IL6R genotype, an association between WC and dietary energy intake level was found in the TT + GT-type subjects (P for linear regression = .048), but not in GG subjects (P for linear regression = .555). In addition, logistic regression analysis revealed that the interaction of IL6R (GG vs TT + GT) genotypes and dietary energy intake levels affected risk for abdominal obesity (P for interaction = .030). We concluded that the IL6R Asp358Ala (T/G) polymorphism appears to interact with energy intake and affect abdominal obesity in Japanese men. The interaction of this genotype and energy intake warrants further study.

Polymorphisms in pro- and anti-inflammatory cytokine genes and susceptibility to atherosclerosis: a pathological study of 1503 consecutive autopsy cases.

Atherosclerosis is a chronic inflammatory disease in the intima of the arterial wall, where cytokines play a crucial role in the pathogenesis of this disease. However, the question of whether or not genetic variations in the cytokine genes could influence the development of atherosclerosis has been poorly investigated. We investigated the relationship of nine common single-nucleotide polymorphisms (SNPs) in tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-10, IL-4 and transforming growth factor (TGF)-beta1 with the atherosclerotic severity in 10 different arteries based on 1503 consecutive autopsies of elderly Japanese subjects registered in the Japanese SNPs for geriatric research (JG-SNP) study. The -1031C allele of TNF-alpha was a significant protective factor for atherogenesis in the carotid, femoral and intracranial arteries [odds ratio (OR): 0.72, 0.73 and 0.70, respectively]. The -511T of IL-1beta and the +29T of TGF-beta1 were significant risk factors for atherogenesis in the subclavian and intracranial arteries (OR: 1.35 and 1.48, respectively). In contrast, conventional risk factors for atherogenesis, such as hypertension and diabetes mellitus, conferred independent risks for almost all arteries. Functional SNPs in TNF-alpha, IL-1beta and TGF-beta1 genes play a role in atherogenesis, although their influences are less pronounced than those of conventional risk factors and appear to be limited to specific arteries in the Japanese elderly.

Interaction of angiotensin I-converting enzyme insertion-deletion polymorphism and daily salt intake influences hypertension in Japanese men.

The contribution of angiotensin I-converting enzyme insertion-deletion polymorphism (ACE I/D) to salt-sensitivity hypertension has been extensively studied by means of salt-loading tests, but whether or not the interaction with daily salt intake affects blood pressure still remains to be clarified. We therefore conducted a cross-sectional study of 284 Japanese male workers (age range, 20-64 years) to examine the effect of ACE I/D genotype and daily salt intake on hypertension. Blood pressure was measured and the ACE I/D was identified by polymerase chain reaction (PCR). Daily salt intake was calculated from a food frequency questionnaire (FFQ). In multivariate analyses, we explored the interaction of ACE I/D and salt intake by means of logistic regression analysis and multiple linear regression analysis. ACE I/D per se was not associated with blood pressure levels or hypertension. ACE I/D interacted with daily salt intake and correlated with hypertension (p for interaction = 0.047). In the ID+II genotype, hypertension was increased by high salt intake (p = 0.005), while in the DD genotype it was not (p = 0.257). The interaction was more prominent in the overweight group (p = 0.039) than in non-overweight group. In the overweight group, high salt intake induced a 10.5 mmHg higher diastolic blood pressure in the ID+II genotype than in the DD genotype (p = 0.042). Our results suggest that ACE I/D and daily salt intake constitute a gene-environment interaction, which may be further modulated by overweight.

Association found between the promoter region polymorphism in the apolipoprotein A-V gene and the serum triglyceride level in Japanese schoolchildren.

The purpose of this study was to assess the influence of single nucleotide polymorphism 3 (SNP3) of the apolipoprotein A-V ( APOA5) gene on the serum triglyceride (TG) level in Japanese schoolchildren. To determine the frequency of the genotype, we analyzed 552 schoolchildren. The frequencies of the T/T, T/C and C/C genotypes of the APOA5 gene were 225 (40.8%), 263 (47.6%) and 64 (11.6%), respectively. The serum TG level was significantly different among the genotypic groups after adjustments for age, gender and obesity index (T/T 71.6+/-34.8 mg/dl, T/C 80.7+/-36.1 mg/dl, C/C 94.4+/-69.4 mg/dl, P<0.0001). The odds ratio (95% confidence interval) for hypertriglyceridemia of the C allele was 2.4 (1.0-6.2). Our data suggested that the T/C promoter region polymorphism of the APOA5 gene appears to be a genetic risk factor for hypertriglyceridemia in Japanese children.