Polymorphisms and haplotype structures in genes for transforming growth factor beta1 and its receptors in familial and unselected breast cancers

Alterations in TGF-beta signaling appear to be associated with an altered risk of developing cancer, including breast cancer. We carried out a case-control study on 8 polymorphisms, including 5 in the TGF-beta1 gene (G-800A, C-509T, Leu10-->Pro, Arg25-->Pro and Thr263-->Ile), a polyalanine polymorphism (9A-->6A) in the TGF-betaRI gene and 2 (G-875A and A-364G) in the TGF-betaRII gene, using samples from 2 different populations, Polish familial and Finnish unselected breast cancer cases, together with ethnically and geographically matched controls. Additionally, familial breast cancer cases with respective controls from Sweden and Germany were studied in the Leu10-->Pro polymorphism, making the total number of familial cases 659. Allele, genotype and haplotype analysis on the TGF-beta1 gene as well as an analysis of the combinations of genotypes of the TGF-beta1 and its receptor genes in each individual were performed. Population differences in the allele and genotype distributions were found from 5 of the polymorphisms and 3 common haplotypes from the TGF-beta1 gene between the Finnish and other populations. However, no statistically significant difference between the breast cancer and healthy control groups was found for any of the 8 polymorphisms nor did the haplotype or genotype combination analysis reach statistical significance. Thus, none of the studied polymorphisms from the TGF-beta1 and its receptor genes was found to influence significantly susceptibility to breast cancer. The possible contribution of 6A/6A homozygosity in the TGF-betaRI gene to breast cancer needs to be confirmed in an independent study.     

An intronic variant of the TGFBR1 gene is associated with carcinomas of the kidney and bladder

TGF-beta signaling is frequently perturbed in many human cancers, including renal cell carcinomas (RCCs) and transitional cell carcinomas (TCCs) of the bladder. Genetic alterations of the TGF-beta type 1 receptor (TGFBR1) may contribute to these perturbations. We therefore examined variations in the TGFBR1 gene by PCR, SSCP and RFLP in carcinomas of the urinary system and in tissues from noncancer, age-matched controls. A G-->A variant 24 bp downstream of the exon/intron 7 boundary of the TGFBR1 gene (Int7G24A) was evident in patients with RCC (46.5%, n = 86) and bladder and upper urinary tract TCC (49.2%, n = 65) significantly more frequently than in age-matched controls (28.3%, n = 113, p < 0.002 by chi2 test). Moreover, 8 homozygous variant carriers were found in the cancer groups, whereas not a single homozygous variant carrier was found in the control group. The Int7G24A allele (both heterozygous G/A and homozygous A/A carriers) was associated with increased RCC incidence (OR = 2.20, 95% CI 1.22-3.96) and TCC incidence (OR = 2.45, 95% CI 1.89-3.16). One somatic mutation of serine to phenylalanine at codon 57 of the TGFBR1 gene was confirmed in an upper urinary tract TCC. In conclusion, the Int7G24A variant in the TGFBR1 gene is significantly more frequent in patients with RCC and TCC than normal age-matched controls, suggesting that it may represent a risk factor for the development of kidney and bladder carcinomas.     

Elevated expression of TGF-beta1 in head and neck cancer-associated fibroblasts

Head and neck cancers are characterized by a vigorous desmoplastic response, but the contribution of stromal-derived growth factors to the tumor microenvironment is poorly understood. We evaluated the expression of stromal growth factor expression in head and neck squamous cell carcinoma (HNSCC) in normal and tumor-associated stromal cells. Stromal tissue was isolated from epithelial cells with laser capture microdissection (LCMD) and analyzed by cDNA array for the expression of TGFalpha, TGF-beta1, HGF, PDGF-alpha, IGFII, bFGF, aFGF, VEGFC, and VEGF. Primary fibroblasts were isolated in vitro from HNSCC tumors, adjacent histologically normal mucosa, and skin in vitro. Fibroblast populations were assessed for TGF-beta1 expression by ELISA and luciferase reporter assay to assess protein expression. We identified TGF-beta1 and IGFII overexpression in normal and tumor-associated stromal cells; however, only TGF-beta1 was significantly overexpressed (3.4-fold) in tumor-associated stroma. Assessment of carcinoma-associated fibroblasts (CAFs), normal dermal fibroblasts (NDFs), and normal mucosal fibroblasts (NMFs) in propagated fibroblasts demonstrated persistently elevated levels of TGF-beta1 in CAFs compared to NMF and NDF populations. Elevated levels of TGF-beta1 were identified in the stromal compartment of HNSCC tumors compared to normal mucosa by immunohistochemical analysis. These results suggest that TGF-beta1 mRNA and protein is specifically upregulated in CAFs in vitro and in vivo.     

Organizational strategy use in obsessive-compulsive disorder

It has been reported that the balance between T-helper type 1 (Th1) cytokines and T-helper type 2 (Th2) cytokines plays a role in psychiatric disorders such as bipolar disorder. The T-helper type 3 (Th3) cytokine, which transforming growth factor beta-1 (TGF-β1), has been shown to modulate the production of Th1 and Th2 cytokines. However, the role of TGF-β1 in bipolar disorder has not yet been explored. A total of 70 manic patients with bipolar disorder and 96 normal controls was recruited. The plasma levels of IFN-γ, IL-4, and TGF-β1 were studied at the time of admission and 8 weeks after mood stabilizer treatment. The detection rate and plasma concentrations of IFN-γ and IL-4 and the IFN-γ/TGF-β1 and IL-4/TGF-β1 ratios were significantly higher in patients than in controls, while the TGF-β1 level was significantly lower. The TGF-β1 level increased significantly after treatment and the IFN-γ/TGF-β1 and IL-4/TGF-β1 ratios returned to control values. TGF-β1 may play a role in the pathophysiology of bipolar disorder through the action of TGF-β1 in modulating the IL-4/TGF-β1 ratio.     

Expression and regulation of nonsteroidal anti-inflammatory drug-activated gene (NAG-1) in human and mouse tissue

BACKGROUND & AIMS: Nonsteroidal anti-inflammatory drugs (NSAIDs) induce NSAID-activated gene 1 (NAG-1), which has proapoptotic and antitumorigenic activities. However, NAG-1 expression and its relationship with apoptosis in human and mouse intestinal tract have not been determined. METHODS: NAG-1 expression in human and mouse tissue was determined by immunohistochemistry, and apoptosis was estimated by in situ apoptosis detection. Apoptosis in NAG-1 overexpressing HCT-116 cells was examined with flow cytometry after cell sorting by green fluorescence protein. NAG-1 regulation in mouse cells was examined by Northern blot analysis, comparing sulindac-treated and nontreated mice. RESULTS: Apoptosis was higher in NAG-1 overexpressing cells compared with controls. Human NAG-1 protein was localized to the colonic surface epithelium where cells undergo apoptosis, and higher expression was observed in the normal surface epithelium than in most of the tumors. This localization and lower expression in tumors was similar to that in the Min mouse, in which NSAIDs were also shown to regulate the expression of NAG-1 in mouse cells. Sulindac treatment of mice increased the NAG-1 expression in the colon and liver. CONCLUSIONS: Based on these results, we propose that NAG-1 acts as a mediator of apoptosis in intestinal cells and may contribute to cancer chemoprevention by NSAIDs.     

<Emphasis Type="Italic">Transforming growth factor B1</Emphasis> T29C polymorphism and breast cancer risk in Japanese women

BACKGROUND: A cohort study for Caucasians aged 65 years or older demonstrated a marked breast cancer risk reduction for those with the CC genotype of transforming growth factor B1 (TGF B1) T29C polymorphism. This is a prevalent case-control study to examine the reported risk reduction for Japanese women. PATIENTS AND METHODS: A total of 232 histologically diagnosed breast cancer patients who visited Aichi Cancer Center Hospital between June 1999 and March 2000 were enrolled. The controls were 172 female outpatients without cancer at the same hospital. DNA was extracted from peripheral blood, and TGF B1 genotype was determined by PCR-CTPP. RESULTS: The genotype frequency was 23.7% for TT, 49.2% for TC, and 27.1% for CC among controls, and 28.9%, 46.1%, and 25.0%, respectively, among cases. Age-adjusted odds ratio (OR) relative to the TT genotype was 0.81 (95% confidence interval, 0.50-1.31) for the TC genotype and 0.77 (0.45-1.34) for the CC genotype. For premenopausal women, the CC genotype was significantly associated with reduced risk of breast cancer in comparison with the TT genotype (OR=0.45, 0.20-0.98). The association was not observed for postmenopausal women (OR=1.40, 0.64-3.08). CONCLUSION: The present study showed risk reduction for Japanese premenopausal women with the CC genotype, but not for postmenopausal Japanese women.     

血清TGFβ1浓度与胃癌侵袭性及其根治手术的关系探讨

探讨ＴＧＦβ１对人胃癌侵袭性的作用。方法应用酶联免吸附法测定２９例胃癌病人血清转化生长因子β１浓度。结果：浸润型组或伴有淋巴结转移组的血清ＴＧＦβ１浓度分别明显高于膨胀型组或无淋巴结转移组,切除瘤体后上述各组的血清ＴＧＦβ１浓度均明显下降。结论：ＴＧＦβ１可能与人胃癌的浸润和转移密切相关。     

Transforming growth factor beta related to extent of tumor angiogenesis but not apoptosis or proliferation in breast carcinoma

Background  Recent investigations have demonstrated the clinical significance of intralesional mean vessel density (ILVD), as a marker of tumor angiogenesis. The role of growth factors in mediating angiogenesis has also been well documented. Transforming growth factor beta (TGF/β) belongs to a family of polypeptides with diverse biological functions. Very few studies however have looked at the role of this growth factor in relation to angiogenesis. This study analyzed the significance of TGFβ in relation to CD34, an endothelial cell marker, the extent of apoptosis, and tissue proliferation defined by Ki67 expression in breast cancer. Methods  The extent of apoptosis was defined by morphological criteria and the Tdt-mediated dUTP biotin nick end labelling (TUNEL) assay. Immunocyfochemistry was performed to measure TGFβ, CD34 and Ki67 expression. Results  An inverse association was observed between TGFβ expression and ILVD as evident by CD34 labelling (r = - 0.311 82, p = 0.00005). TGFβ expression did not correlate with either TUNEL reactivity or Ki67 expression. CD34 and TGFβ expression also had no relationship with histopathological grade. No correlation was observed between CD34 expression and apoptosis. However a statistically significant correlation was observed between CD34 and Ki67 expression. Conclusions  These results suggesl that breast cancer cells synthesize TGFβ that, through paracrine mechanisms, may inhibit proliferation of vascular endothelium rather than their own growth Moreover the data also suggest that decreased expression of TGFβ was associated with on increase in neovascularization, which in turn would increase the tumor proliferative fraction.     

EGFR status and EGFR ligand expression influence the treatment response of head and neck cancer cell lines

Background: Combination treatment (chemoradiotherapy) is the standard treatment for locally advanced head and neck squamous cell carcinoma (HNSCC); however, treatment resistance and local recurrence are significant problems. A high level of epidermal growth factor receptor (EGFR) has been associated with a more aggressive phenotype as well as decreased responsiveness to radio‐ or chemotherapy. We examined the role of EGFR status and EGFR ligand expression for the treatment response. Methods: Intrinsic sensitivity to radiotherapy, cisplatin, and cetuximab treatments was investigated in 25 HNSCC cell lines. EGFR gene copy number, mRNA and protein expression, EGFR and Akt phosphorylation status, and mRNA expression of the EGFR ligands were analyzed using quantitative PCR and ELISA and assessed for their impact on treatment sensitivity. Results: Different treatment modalities yielded great diversity in outcome; of note, cetuximab treatment stimulated growth in one cell line. When treatments were combined primarily additive effects were observed. While radioresistance tended to be associated with a high level of phosphorylated EGFR (pEGFR; P = 0.09), cetuximab‐resistant cells had low levels of pEGFR (P = 0.13). The three most cetuximab‐sensitive cell lines had high EGFR gene copy numbers. Furthermore, cetuximab treatment response was significantly correlated with epiregulin mRNA expression (r = ?0.408, P = 0.043). Cisplatin‐resistant tumor cells expressed significantly lower levels of EGFR protein (P = 0.04) compared to cisplatin‐sensitive cells and tended to have lower levels of phosphorylated Akt (pAkt; P = 0.13) and lower expression levels of amphiregulin (P = 0.18). Conclusions: Epidermal growth factor receptor status and ligand expression influence the treatment sensitivity of HNSCC cells and may be useful as predictive markers.