Prostate cancer stromal cells and LNCaP cells coordinately activate the androgen receptor through synthesis of testosterone and dihydrotestosterone from dehydroepiandrosterone

金沢大学附属病院泌尿器科金沢大学附属病院泌尿器科One of the mechanisms through which advanced prostate cancer (PCa) usually relapses after androgen deprivation therapy (ADT) is the adaptation to residual androgens in PCa tissue. It has been observed that androgen biosynthesis in PCa tissue plays an important role in this adaptation. In the present study, we investigated how stromal cells affect adrenal androgen dehydroepiandrosterone (DHEA) metabolism in androgen-sensitive PCa LNCaP cells. DHEA alone had little effect on prostate-specific antigen (PSA) promoter activity and the proliferation of LNCaP cells. However, the addition of prostate stromal cells or PCa-derived stromal cells (PCaSC) increased DHEA-induced PSA promoter activity via androgen receptor activation in the LNCaP cells. Moreover, PCaSC stimulated the proliferation of LNCaP cells under physiological concentrations of DHEA. Biosynthesis of testosterone or dihydrotestosterone from DHEA in stromal cells and LNCaP cells was involved in this stimulation of LNCaP cell proliferation. Androgen biosynthesis from DHEA depended upon the activity of various steroidogenic enzymes present in stromal cells. Finally, the dual 5a-reductase inhibitor dutasteride appears to function not only as a 5a-reductase inhibitor but also as a 3b-hydroxysteroid dehydrogenase inhibitor in LNCaP cells. Taken together, this coculture assay system provides new insights of coordinate androgen biosynthesis under the microenvironment of PCa cells before and after ADT, and offers a model system for the identification of important steroidogenic enzymes involved in PCa progression and for the development of the corresponding inhibitors of androgen biosynthesis. © 2009 Society for Endocrinology

ベトナムダイオキシン高濃度汚染地域における授乳中の母親の唾液中ホルモン値の検討

【目的】アメリカ軍は１９６１年から１９７１年の間に８０００万リットル以上の枯葉剤を南ベトナムに撒布した。この研究の目的はベトナムの枯葉剤高濃度汚染地域（Binh Dinh省Phu Cat県）と非汚染地域（Ha Nam省Kim Bang県）において枯葉剤（オレンジ剤）の暴露と人体への健康影響との関連を検討することである。【方法】２つの地域において疫学的調査を実施した。対象者は授乳中で生後４週から１６週の乳児を持つ２０－３０歳の母親とし、汚染地域では５８名、非汚染地域では５３名であった。疾患の危険因子等に関する情報は母親への面接調査から得た。母乳をすべての対象者から採取したが、唾液を採取したのは汚染地域４１名、非汚染地域１９名であった。母親と子どもの身体計測を行い２地域間で比較を行った。【結果と考察】汚染地域における乳児の体重と胸囲は有意に非汚染地域よりも小さいが、年齢(週数)も汚染地域の方が有意に低かった。汚染地域における母親とその家族の現病歴は非汚染地域よりも有意に多かった。母親の視力は両眼とも２地域間で有意差は認められなかった。汚染地域と非汚染地域双方の母親の唾液中のコルチゾン値は、母乳中のコルチゾン値と高い相関を示した。汚染地域と非汚染地域間で唾液中のホルモン値には有意差は認められなかった。Objective : Between 1961 and 1971 the US military used over 80 million litres of herbicides in southern Vietnam. This study aims to assess hormone levels in the saliva of lactating Vietnamese mothers and human health effects in a dioxin hot spot (Phu Cat district, Binh Dinh province) and a non-exposed (Kim Bang district, Ha Nam province) area in Vietnam. Materials and Methods : An epidemiological study was carried out in both areas. The subjects were 58 lactating females in the hot spot area and 53 lactating females in the non-exposed area. All were aged between 20 and 30 years with infants aged between 4 and 16 weeks. Information about disease risk factors was obtained through interviews with mothers. Breastmilk samples were taken from all subjects, whereas saliva samples were obtained from 41 mothers in the hot spot area and 19 in the non-exposed area. Body measurements for both mothers and their infants were compared between the two areas. Result and Discussion : The weight and chest circumference of infants in the hot spot area were significantly lower than those in the non-exposed area, whereas age (weeks) is also significantly younger in hot spot area than non-exposed area. Present maternal and family diseases in the hot spot area were significantly higher than those in the non-exposed area. Maternal eyesight in both eyes did not differ significantly between the two areas. The cortisone levels in saliva have been found to be closely related to those in breast milk samples of both mothers in hot spot and non-exposed areas. No significant difference was found between salivary hormone levels of mothers in the hot spot and non-exposed areas

Inverse association of highly chlorinated dioxin congeners in maternal breast milk with dehydroepiandrosterone levels in three-year-old Vietnamese children

This study aims to evaluate the endocrine-disrupting effect of dioxin congeners on adrenal steroid hormones in mother-child pairs. In our previous study, we found that cortisol and cortisone levels were higher in the blood and the saliva of mothers living in a dioxin hotspot area than in mothers from a non-exposed region in Vietnam. In this follow-up study, we determined the salivary steroid hormone levels in 49 and 55 three-year-old children of these mothers in the hotspot and non-exposed region, respectively. Steroid hormones were determined by liquid chromatography-tandem mass spectrometry, and dioxin in the maternal breast milk was determined by gas chromatography-mass spectrometry. Dioxin levels in the breast milk of mothers from the hotspot (median total toxic equivalents polychlorinated dibenzodioxins/polychlorinated dibenzofurans; (TEQ PCDD/Fs) of 11. pg/g lipid) were three to four times higher than those of mothers in the non-exposed region (median TEQ PCDD/Fs of 3.07. pg/g lipid). Salivary dehydroepiandrosterone (DHEA) levels in children were found to be significantly lower in the hotspot than in the non-exposed region, while cortisol and cortisone levels were not different between the two regions. Highly chlorinated dioxin congeners, such as octacholorodibenzodioxin (OCDD), 1,2,3,4,6,7,8-heptacholorodibenzodioxin (HpCDD) and 1,2,3,4 (or 6), 7,8-hexachlorodibenzodioxin Hx(CDD), showed stronger inverse associations with the children\u27s salivary DHEA than other lowly chlorinated dioxin congeners. Glucocorticoid levels in the mothers exhibited a significantly positive correlation with OCDD and HpCDD/F (polychlorinated dibenzofurans). In conclusion, highly chlorinated dioxin congeners are more strongly correlated with endocrine-disrupting effects on adrenal hormones, resulting in high cortisol levels in the mothers and low DHEA levels in their three-year-old children. © 2016 Elsevier B.V.Embargo Period 24 month

Influence of dioxin exposure upon levels of prostate-specific antigen and steroid hormones in Vietnamese men

Most studies on the relationship between Agent Orange and prostate cancer have focused on US veterans of the Vietnam War. There have been few studies focusing on the relationship between levels of prostate-specific antigen (PSA) and dioxins or steroid hormones in Vietnamese men. In 2009–2011, we collected blood samples from 97 men who had resided in a “dioxin hotspot” and 85 men from a non-sprayed region in Vietnam. Then levels of PSA, dioxins, and steroid hormones were analyzed. Levels of most dioxins, furans, and non-ortho polychlorinated biphenyls were higher in the hotspot than those in the non-sprayed region. Levels of testosterone, dehydroepiandrosterone, and estradiol differed significantly between the hotspot and the non-sprayed region, but there were no correlations between levels of PSA and steroid hormones and dioxins in either of the two regions. Our findings suggest that PSA levels in Vietnamese men are not associated with levels of dioxin or steroid hormones in these two regions. © 2016 Springer-Verlag Berlin HeidelbergEmbargo Period 12 month

High cortisol and cortisone levels are associated with breast milk dioxin concentrations in Vietnamese women

Objective: Dioxin (polychlorinated dibenzodioxins Cpolychlorinated dibenzofurans) is one of the most toxic chemical substances known. Although it is suspected to cause endocrine disruption, very few epidemiological studies have been carried out on its effects on human steroid hormones. The aim of this study was to elucidate the association of dioxin exposure with steroid hormone levels in the saliva and serum of Vietnamese women. Study design : Two areas, namely Phu Cat (hot spot) and Kim Bang (nonexposed area), were selected for the study. The study subjects consisted of 51 and 58 women respectively. Saliva, blood, and breast milk samples were collected from the subjects in both the areas. Methods: Cortisol, cortisone, DHEA, androstenedione, estrone, and estradiol levels in serum and saliva were determined by liquid chromatography-tandem mass spectrometry; dioxin concentrations in breast milk were measured by gas chromatography-mass spectrometry. Results: Dioxin concentrations in the breast milk of women from the dioxin hot spot were three to four times higher than those in the breast milk of women from the nonexposed area. Good correlations were found between the levels of six steroid hormones in saliva and those in serum respectively. Salivary and serum cortisol and cortisone levels in women from the dioxin hot spot were significantly higher than those in women from the nonexposed area (P<0.001) and those in all the subjects were positively associated with dioxin concentrations in Vietnamese women (P<0.01). Conclusion: These results suggest that dioxin influences steroidogenesis in humans. Saliva samples can be used for hormone analysis and are therefore excellent specimens in epidemiological studies. © 2014 European Society of Endocrinology

Low birth weight of Vietnamese infants is related to their mother’s dioxin and glucocorticoid levels

We aimed to determine the relationship between dioxin congeners in maternal breast milk and maternal glucocorticoid levels with newborn birth weight after nearly 45 years of use of herbicides in the Vietnam War. The study subjects comprised 58 mother–infant pairs in a region with high dioxin levels in the soil (hotspot) and 62 pairs from a control region. Dioxin levels in maternal breast milk were measured by HRGC-HRMS. Salivary glucocorticoid levels were determined by LC-MS/MS. Dioxin congener levels in mothers from the hotspot were found to be two to fivefold higher than those in mothers from the control region. Birth weight was inversely correlated with 2,3,7,8-TeCDD and 2,3,4,7,8-PeCDF congener levels. The rate of newborns whose birth weight was less than 2500 g was threefold higher in the hotspot (12 %) than in the control region (4 %). Salivary glucocorticoid levels in mothers with low birth weight infants were significantly higher than those in the normal birth weight group. Low birth weight of Vietnamese newborns in a hotspot for dioxin levels is related to some dioxin congener levels and high glucocorticoid levels in mothers. This finding in mother–infant pairs suggests that excess maternal glucocorticoid levels are related to dioxin burden and they result in low birth weight. © 2016 Springer-Verlag Berlin HeidelbergEmbargo Period 12 month

Levels of polychlorinated dibenzodioxins and polychlorinated dibenzofurans in breast milk samples from three dioxin-contaminated hotspots of Vietnam

We determined polychlorinated dibenzodioxin (PCDD) and polychlorinated dibenzofuran (PCDF) levels in breast milk of 143 primiparae living around the three most dioxin-contaminated areas of Vietnam. The women sampled lived in the vicinity of former U.S. air bases at Bien Hoa (n. =. 51), Phu Cat (n. =. 23), and Da Nang (n. =. 69), which are known as dioxin hotspots. Breast milk samples from Bien Hoa City, where residents live very close to the air base, showed high levels of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), with 18% of the samples containing >. 5. pg. TCDD/g lipid. However, Phu Cat residents lived far from the air base and their samples showed lower TCDD levels, with none containing >. 5. pg. TCDD/g lipid. In Da Nang, TCDD levels in mothers from Thanh Khe (close to the air base, n. =. 43) were significantly higher than those in mothers from Son Tra (far from the air base, n. =. 26), but not other PCDD and PCDF (PCDD/F) congeners. Although TCDD levels in Bien Hoa were the highest among these hotspots, levels of other PCDD/F congeners as well as the geometric mean concentration of total PCDD/F level in Bien Hoa (9.3. pg toxic equivalents [TEQ]/g lipid) were significantly lower than the level observed in Phu Cat (14.1. pg. TEQ/g lipid), Thanh Khe (14.3. pg. TEQ/g lipid), and Son Tra (13.9. pg. TEQ/g lipid). Our findings indicated that residents living close to former U.S. air bases were exposed to elevated levels of TCDD, but not of other PCDD/F congeners

High testosterone levels in prostate tissue obtained by needle biopsy correlate with poor-prognosis factors in prostate cancer patients

Background: There is currently no consensus on the correlations between androgen concentrations in prostate tissue and blood and stage and pathological grade of prostate cancer. In this study, we used a newly-developed ultra-sensitive liquid-chromatography tandem mass spectrometry method to measure testosterone (T) and dihydrotestosterone (DHT) concentrations in blood and needle biopsy prostate specimens from patients with prostate cancer.Methods: We analyzed androgen levels in 196 men diagnosed with prostate cancer. All patients had undergone systematic needle biopsy, and an additional needle biopsy from the peripheral zone was conducted for the simultaneous determination of T and DHT. We analyzed the relationships between T and DHT levels in tissue and blood and Gleason score, clinical stage, and percentage of positive biopsy cores, using multivariate analysis. Results: The median T and DHT levels in blood were 3551.0 pg/mL and 330.5 pg/mL, respectively. There was a strong correlation between serum T and DHT. The median T and DHT levels in prostate tissue were 0.5667 pg/mg and 7.0625 pg/mg, respectively. In multivariate analysis, serum prostate-specific antigen and tissue T levels were significantly associated with poor prognosis; high T levels in prostate tissue were significantly related to high Gleason score (p = 0.041), advanced clinical stage (p = 0.002), and a high percentage of positive biopsy cores (p = 0.001). Conclusions: The results of this study indicate that high T levels in prostate tissue are related to high Gleason score, advanced clinical stage, and a high percentage of positive biopsy cores in patients with prostate cancer. T level in needle biopsy specimens may therefore be a useful prognostic factor in prostate cancer patients