Her-2/neu amplification and breast cancer survival: results from the Shanghai breast cancer study.

Her-2/neu is a member of the epidermal growth factor receptor family that has been found to be overexpressed or amplified in approximately 20-30% of breast cancers. Negative prognosticators and a shortened survival have been shown to be associated with these changes in Her-2/neu, but previous studies have consisted of predominantly Caucasian populations. Additionally, chromogenic in situ hybridization (CISH) has been suggested to be a potential alternative to fluorescent in situ hybridization (FISH), the expensive and labor-intensive gold standard assay currently used for Her-2/neu amplification. This study evaluated breast cancer samples from 313 Chinese women participating in the Shanghai breast cancer study, of which 100 (32%) were found to have Her-2/neu amplification by either FISH or CISH methodologies. After a mean follow-up period of 6.67 years, Her-2/neu amplification was found to be significantly associated with an increased hazard of death, regardless of which assay was used to detect amplification. Patients with Her-2/neu amplification were approximately 60% more likely to die of the disease (HR: 1.6, 95% CI: 1.0-2.6) than patients without amplification, even after adjusting for age, stage, menopausal status, chemotherapy, radiotherapy and tamoxifen treatment. Furthermore, the negative prognostic effect of Her-2/neu varied by cancer stage, with greater risks of death evident among later stage patients. This study supports a negative prognostic role for Her-2/neu in breast cancer survival among a Chinese population, irrespective of whether FISH or CISH is used to detect amplification of the Her-2/neu gene.

Haplotype analyses of CYP19A1 gene variants and breast cancer risk: results from the Shanghai Breast Cancer Study.

Estrogens play a central role in the etiology of breast cancer. The CYP19A1 gene encodes aromatase, a key enzyme in the biosynthesis of estrogens. Several single nucleotide polymorphisms (SNP) or haplotypes in the CYP19A1 gene have been evaluated in relation to breast cancer risk. However, the results have been inconsistent. In this study, we constructed haplotypes of the CYP19A1 gene using 19 haplotype-tagging SNPs in Chinese women and evaluated the variation of this gene in relation to breast cancer risk in a population-based case-control study involving 1,140 cases and 1,244 community controls of the Shanghai Breast Cancer Study. Five common haplotypes in block 1, three common haplotypes in block 2, five common haplotypes in block 3, and four common haplotypes in block 4 were identified. No apparent association was observed between common haplotypes and breast cancer risk in analyses including all subjects nor in analyses stratified by menopausal status. Similarly, no statistically significant differences were found between cases and controls in the genotype distributions of the 19 individual SNPs and the (TTTA)(n) repeat polymorphism evaluated in the study. No overall association of breast cancer risk with common CYP19A1 gene variants among Chinese women was observed in this large-scale, comprehensive study. Further studies are needed to explore CYP19A1 gene-environment interactions in relation to breast cancer risk.

Polymorphisms in the CYP19A1 (aromatase) gene and endometrial cancer risk in Chinese women.

Aromatase, encoded by the CYP19A1 gene, is a key enzyme in estradiol biosynthesis, which catalyzes the conversion of androstenedione and testosterone to estrone and estradiol, respectively. Given the critical role of estrogen in the development of endometrial cancer risk, we evaluated genetic polymorphisms of the CYP19A1 gene, including rs1065779, rs700519, rs28566535, rs752760, and rs1870050, in association with endometrial cancer in a population-based case-control study conducted in Shanghai, China. Genotypes of 1,040 incident endometrial cancer cases and 1,031 frequency-matched controls were included in the study. We applied a logistic regression model to derive adjusted odds ratios (OR) and their 95% confidence intervals (95% CI). Six common haplotypes with a frequency >or=5% were estimated; the highest frequency haplotype was GCACA (27.8% in cases and 26.2% in controls). We observed an inverse association between CYP19A1 haplotype TCATC and endometrial cancer in our population (OR, 0.76; 95% CI, 0.62-0.92). An inverse association was found between endometrial cancer and single nucleotide polymorphism rs1870050 in the promoter region with ORs of 0.81 (95% CI, 0.68-0.97) and 0.58 (95% CI, 0.42-0.80) for the AC and CC genotypes, respectively. We observed a multiplicative interaction between single nucleotide polymorphism rs700519 and body mass index among postmenopausal women (P = 0.01), with stronger associations between rs700519 genotypes and endometrial cancer risk among heavier (body mass index, >or=25) postmenopausal women. In summary, our data show that polymorphisms in the CYP19A1 gene may contribute to endometrial carcinogenesis.

Association of endometrial cancer risk with a functional polymorphism (Asp(327)Asn) in the sex hormone-binding globulin gene.

BACKGROUND: Sex steroid hormones play a central role in the development of endometrial cancer. Sex hormone-binding globulin (SHBG) modulates the bioavailability of circulating sex hormones and regulates their signaling systems in target cells. Epidemiologic studies have demonstrated that blood SHBG levels are associated inversely with the risk of endometrial cancer. A functional polymorphism, an amino acid substitution of asparagine (Asp) for aspartic acid (Asn) at residue 327 (Asp(327)Asn) (reference sequence 6259), in the SHBG gene recently was identified and has been associated with an increased half-life and elevated blood levels of SHBG. The authors tested the hypothesis that this genetic variance is associated with a reduced risk of endometrial cancer in the Shanghai Endometrial Cancer Study, a population-based, case-control study that was conducted in urban Shanghai, China between 1997 and 2003. METHODS: This study included 1037 women with newly diagnosed endometrial cancer ages 30 years and 69 years and 1031 age-matched controls from the community who had completed an in-person interview and donated a blood and/or buccal cell sample to the study. Genotyping for the Asp(327)Asn polymorphism was performed by using the TaqMan method. Odds ratio (ORs) and 95% confidence intervals (95% CIs) derived from logistic regression models were used to evaluate the association between the genetic variation and endometrial cancer risk. RESULTS: The allele frequencies of Asn were 15.0% in cases and 17.1% in controls (P = .06). The variant Asn(327) allele was associated with a reduced risk of endometrial cancer in postmenopausal women (OR, 0.72; 95% CI, 0.55-0.93) but not in premenopausal women (OR, 1.02; 95% CI, 0.74-1.42). The inverse association was more pronounced among postmenopausal women who had a low body mass index (OR, 0.48; 95% CI, 0.31-0.76) or longer years of menstruation (OR, 0.64; 95% CI, 0.46-0.89), although the results from tests for multiplicative interaction were not statistically significant. CONCLUSIONS: The current results suggested that the codon 327 Asn allele in the SHBG gene may be related to a reduced risk of endometrial cancer among postmenopausal women.

E-cadherin promoter polymorphisms are not associated with the aggressiveness of prostate cancer in Caucasian patients.

BACKGROUND: -160C-->A and -347G-->GA polymorphisms in the promoter region decrease E-cadherin gene transcription. Decreased E-cadherin expression predicts poor outcome among patients with cancer. We sought to investigate whether -160C-->A and/or -347G-->GA polymorphisms were associated with the aggressiveness of prostate cancer. METHODS: TaqMan single nucleotide polymorphism genotyping assay (Applied Biosystems, Foster City, CA) was used to detect -160C-->A and -347G-->GA polymorphisms in deoxyribonucleic acid from the paraffin-embedded prostate tissues of 98 Caucasian patients. RESULTS: The genotype frequencies were -160C/C: 48% (47 of 98); -160C/A: 44% (43 of 98); -160A/A: 8% (8 of 98); -347G/G: 68% (67 of 98); -347G/GA: 28% (27 of 98); and -347GA/GA: 4% (4 of 98). Using the chi-square test, we found that the polymorphisms -160C-->A and -347G-->GA were not related to other clinical and pathologic parameters (i.e., age, prostate-specific antigen level, Gleason grade, and clinical stage) (P > 0.05). In combination analysis, there was no significant relationship between patients with both -160C/C and -347G/G, and these same parameters (P > 0.05). Using the log-rank test, we found no significant difference in relapse-free survival and overall survival between patients with -160C/C and those with -160A/C or -160A/A (P = 0.0764 and 0.2746, respectively), and also no significant difference between patients with -347G/G and those with -347GA/G or -347GA/GA (P = 0.9416 and 0.7367, respectively). There was also no significant difference in relapse-free survival and overall survival between patients with homozygosities of -160C/-347G and patients with other genotypes (P = 0.1418 and 0.2434, respectively). CONCLUSION: We conclude that E-cadherin -160C-->A and/or -347G-->GA polymorphisms are not associated with the aggressiveness of prostate cancer in Caucasian patients.

Genetic polymorphisms of the CYP19A1 gene and breast cancer survival.

The CYP19A1 protein (aromatase) plays a critical role in estrogen biosynthesis and thus may be related to the progression of breast cancer. We examined the association between CYP19A1 genetic polymorphisms and breast cancer survival in a cohort of 1,136 patients who were recruited as part of a population-based case-control study in Shanghai, China from 1996 to 1998 and who has donated a DNA sample to the study. Patients were followed for cancer recurrence and mortality through July 2005. Nineteen haplotype tagging single-nucleotide polymorphisms (SNP) in the CYP19A1 gene were evaluated. For each of the five SNPs located in haplotype block 2, patients homozygous for the minor alleles had a reduced 5-year disease-free survival rate compared with those carrying the major allele. The age-adjusted hazard ratios (HR) and 95% confidence intervals (95% CI) were 1.5 (1.1-2.1), 2.1 (1.2-3.6), 1.5 (1.1-2.0), 1.4 (1.0-2.0), and 1.4 (1.0-2.0) for hCV1664178, rs12900137, rs730154, rs936306, and rs1902586, respectively. Haplotype analyses showed that the haplotype CCCTA (all minor alleles of the five SNPs in block 2) was associated with decreased disease-free survival (HR, 1.9; 95% CI, 1.1-3.3). The nonsynonymous SNP, rs700519 (Arg264Cys), located in haplotype block 4, was also associated with breast cancer survival. The age-adjusted HR for the Cys/Cys (T/T) genotype was 2.2 (95% CI, 1.2-4.1) for overall survival and 2.1 (95% CI, 1.1-3.9) for disease-free survival, compared with those carrying the Arg (C) allele. These results suggest that polymorphisms in the CYP19A1 gene may have effects on breast cancer prognosis.

Immunohistochemical expressions of Ki-67, cyclin D1, beta-catenin, cyclooxygenase-2, and epidermal growth factor receptor in human colorectal adenoma: a validation study of tissue microarrays.

BACKGROUND: Tissue microarray (TMA) holds promise as a high-throughput method for the analysis of biomarkers in tissue specimens. The validity and reliability of this method, however, may vary for different biomarkers in different tissue specimens. OBJECTIVES: In this study, we evaluated the validity and reliability of using TMA to assess biomarkers in colorectal adenomas. METHODS: Sixty-three consecutive patients with colorectal adenomas were recruited in this study. Two TMA blocks were constructed using four punches from each adenoma (one periphery, one deep, and two middle zones). The immunostaining of five markers (Ki-67, cyclin D1, beta-catenin, cyclooxygenase-2, and epidermal growth factor receptor) was analyzed, and the concordance between data obtained from TMAs and standard whole-tissue sections was evaluated by Spearman's correlation and kappa analysis. RESULTS: Colorectal adenoma exhibited zonal, heterogeneous expression patterns for all five markers. The concordance rates for the semiquantitative evaluation of markers between data from TMAs and whole sections ranged from 87% to 93% with corresponding kappa statistics of 77% to 90%. In addition, both quantitative and semiquantitative methods were used to score TMA sections, and good correlations between these two methods were shown for all five markers with intraclass correlation coefficients ranging from 0.5 to 0.8. CONCLUSION: Our study indicates that TMA can be used to reliably assess the expression levels of Ki-67, cyclin D1, beta-catenin, cyclooxygenase-2, and epidermal growth factor receptor in colorectal adenoma tissues.

Population-based case-control study of VEGF gene polymorphisms and breast cancer risk among Chinese women.

Vascular endothelial growth factor (VEGF) is a major angiogenic factor involved in a number of pathologic processes, including neovascularization, a crucial step in the development of solid malignancies. Using data and specimens collected in the Shanghai Breast Cancer Study, a population-based case-control study conducted in urban Shanghai, China from 1996 to 1998, we evaluated the association of VEGF gene polymorphisms with breast cancer risk. Included in this study were 1,093 cases and 1,184 age-matched controls who had completed an in-person interview and donated a blood sample to the study. Polymorphisms in the promoter region (T -460C), 5' untranslated region (C +405G), and 3'untranslated region (C936T) were genotyped using the Taqman allelic discrimination assay. No statistically significant case-control difference was found for the C +405G and T -460C polymorphisms. However, the C936T polymorphism was associated with a reduced risk of breast cancer. Compared with CC genotype carriers, women who had the TT genotype showed a decreased risk [odds ratio (OR), 0.65; 95% confidence interval (95% CI) 0.41-1.02], and the inverse association was restricted to premenopausal women (OR, 0.45; 95% CI, 0.25-0.79). Six common haplotypes were identified. Compared with the most common haplotype (-460T/405C/936C), the -460T/405G/936T haplotype was associated with a reduced risk of breast cancer (OR, 0.67; 95% CI, 0.43-1.04), particularly in premenopausal women (OR, 0.47; 95% CI, 0.27-0.81). Our study suggests that the VEGF C936T polymorphism might be a susceptibility factor for breast cancer among Chinese women.

Association of genetic polymorphisms in the VEGF gene with breast cancer survival.

The vascular endothelial growth factor (VEGF) is an important regulator of angiogenesis and vascular permeability. VEGF overexpression has been associated with advanced stage and poor survival of several cancers. We evaluated the association of functional polymorphisms in the VEGF gene with breast cancer survival in a cohort of 1,193 breast cancer patients who were recruited as part of a population-based case-control study in Shanghai, China from 1996 to 1998 and followed for cancer recurrence and mortality between March 2000 and December 2002. Included in the study were three functional polymorphisms (C-460T, G+405C, and C+936T) in the VEGF gene. Carrying the -460C or +405G allele was associated with decreased overall survival. The age-adjusted hazard ratios (HR) were 1.5 [95% confidence interval (95% CI), 0.9-2.5] for -460CC genotype carriers and 1.6 (95% CI, 1.0-2.5) for +405GG genotype carriers compared with noncarriers. Further analyses showed that the -460T/+450C/+936C haplotype was related to increased survival (HR, 0.57; 95% CI, 0.4-0.9), whereas the -460C/+405G/+936T haplotype was associated with nonsignificantly decreased survival (HR, 2.1; 95% CI, -0.9 to 4.7). The C+936T polymorphism alone was not related to overall or disease-free survival. This study suggests that VEGF polymorphisms may be a significant genetic marker for breast cancer prognosis.

FISH analysis of BRCA1 copy number in paraffin-embedded ovarian cancer tissue samples.

We investigated the BRCA1 gene copy number in unselected ovarian malignancies. Both additional genes (amplification) as well as deletion (loss of heterozygosity, LOH) are often thought to have a role in the initiation or progression of cancer. In addition, if there were little change, deletion studies might help identify BRCA1 mutation carriers. Forty-seven paraffin-embedded ovarian tissue blocks obtained between 1984 and 1997 were used for this study. A sample was "deletion-positive" when BRCA1-deleted cells in the tumor area were significantly different from the benign area. Twenty-five (53%) cases were found to be "deletion-positive". The average age of onset of "deletion-positive" patients was 50.8 years and of "deletion-negative" 57.8 years (P < 0.05). There was no statistical difference between groups in the staging, histology, or prognosis. A Kaplan-Meier study did show a trend towards poorer survival for "deletion-positive" patients. FISH permits unique molecular characterization of malignancies at a cellular level. Double amplification of HER-2 and c-myc predicts poor ovarian cancer survival. There appears to be a definite role for BRCA1 deletion in reducing the age of ovarian cancer onset and possibly in overall survival. Further FISH studies of this and other patient sets using additional molecular markers are needed.