Jenni Bernoulli

Inflammation and epithelial alterations in rat prostate: impact of the androgen to oestrogen ratio.

Chronic non-bacterial prostatitis may offer new insights into the pathogenesis of human benign prostatic hyperplasia and prostate cancer and the strategies for their treatment and prevention. The potential significance of androgen replacement therapy in terms of the reversal of oestradiol (E(2))-induced inflammatory reaction was studied in the dorsolateral prostate (DLP) of the Noble rat. Castrated Noble rats were treated with E(2) and different doses of androgens [dihydrotestosterone (DHT) and testosterone (T)] to achieve an elevated concentration of E(2) and a wide range of the androgen-to-oestradiol ratios in serum. After the 3-week treatment, inflammatory changes in the DLP were classified and counted. Oestrogen receptor alpha (ER alpha), progesterone receptor (PR), fos-related antigen-2 (Fra2), Ki-67 and P63 were immunocytochemically stained. T, E(2) and prolactin concentrations in serum were measured and the relative weights of the seminal vesicles and pituitary glands and microscopic structures of the DLP and seminal vesicle ducts were determined. Hypoandrogenic doses of DHT (judged on the basis of seminal vesicle weight gain), dose-dependently increased the number of perivascular and stromal inflammatory infiltrates. T and DHT were anti-inflammatory at the doses which normalized or over stimulated the growth of the seminal vesicles. As signs of anti-oestrogenicity, androgens dose-dependently decreased the number and distribution of the ER alpha and PR-positive cells at proinflammatory concentrations. Anti-inflammatory concentrations were needed to reduce the expression of Fra2, E(2)-increased prolactin concentration in serum and pituitary weight. The androgen concentrations required to prevent proinflammatory and epithelial responses to E(2) in the presence of elevated E(2) concentrations may subject the accessory sex glands to more intense androgenic stimulation than is normal for the male. The androgen-resistant endpoints of oestrogen action (body weight reduction and hyperplasia of seminal vesicle ducts) further indicate limitations in the possible preventive effects of androgen-replacement therapy.

Prostatic inflammation and obstructive voiding in the adult noble rat: Impact of the testosterone to estradiol ratio in serum

The age‐related decline of the testosterone to estradiol (T‐to‐E2) ratio in serum is associated with the increased prevalence of prostatic inflammation and lower urinary tract symptoms suggesting obstructive voiding. The impact of the T‐to‐E2 ratio on the development and reversal of non‐bacterial prostatic inflammation and obstructive voiding was tested in adult Noble rats.

Histopathological evidence for an association of inflammation with ductal pin‐like lesions but not with ductal adenocarcinoma in the prostate of the noble rat

Chronic inflammation may contribute to the development of prostate cancer. The goal of this study was to determine the possible association of prostatic inflammation, prostatic intraepithelial neoplasia (PIN)‐like lesion, and prostate cancer, and to assess the androgen and estrogen dependency of the early steps of carcinogenesis.

The Soy Effect in the Disease Models of Nonbacterial Prostatitis and Obstructive Voiding

The goal of this study was to improve the understanding of the potential significance of dietary soy for human health by investigating its effects in the animal models of nonbacterial prostatitis and urethral obstruction. Nonbacterial prostatitis was induced in adult Noble rats with the combined treatment of testosterone and 17β-estradiol. The inflammatory foci categorized into three forms were counted and correlated with expression of an estrogen-responsive gene, progesterone receptor (PR), in the dorsolateral lobes of the rats on soy (+) and soy (−) diets. Development of obstructive voiding after neonatal estrogenization of Noble rats (NeoDES rats) was followed with urodynamic measurements in rats on soy (+) and soy (−) diets. The amounts of genistein and daidzein, two major soy-derived isoflavones, were measured in the urine of Noble rats by the high-performance liquid chromatography–photodiodearray method. Dietary soy decreased the total number of inflammatory foci while no demonstrable effects were seen on the cellular composition of the infiltrates. Soy did not increase the weights of the pituitary gland, testes, or sex accessory glands, but it did increase the number of PR-positive epithelial cells in the dorsolateral prostate. It also decreased the bladder pressures in NeoDES rats but did not increase the flow rates. The soy effects may be mediated by the strong estrogen influence involved in the animal models. Dietary soy had anti-inflammatory effects in the prostate but only marginal effects on the development of obstructive voiding in Noble rats. The anti-inflammatory effects of soy may contribute to the lower prevalence of prostatitis-like symptoms and the historically lower risk of benign prostatic hyperplasia in Japan; however, no evidence was found that regular consumption of soy influences the age-related development of lower urinary tract symptoms or decline of flow rate.

Fispemifene [Z-2-{2-[4-(4-Chloro-1,2-diphenylbut-1-enyl)-phenoxy]ethoxy}-ethanol], a Novel Selective Estrogen Receptor Modulator, Attenuates Glandular Inflammation in an Animal Model of Chronic Nonbacterial Prostatitis

The anti-inflammatory and antiestrogenic action of fispemifene [Z-2-{2-[4-(4-chloro-1,2-diphenylbut-1-enyl)phenoxy]ethoxy}-ethanol], a novel selective estrogen receptor modulator (SERM), was tested on the Noble rat model of chronic nonbacterial prostatic inflammation with cellular composition and inflammation patterns similar to those described in human prostatitis. Inflammation was assessed by counting perivascular and stromal infiltrates and the number of inflamed acini. Furthermore, the aggressiveness of inflammation was assessed on the basis of the relation of lymphocytes to the acinar epithelium. The immunohistochemical expression of progesterone receptor (PR) and Fos-related antigen 2 (Fra2), prolactin concentration in serum, and the weights of the seminal vesicles and pituitary glands were used as endpoints of estrogen action. Fispemifene significantly attenuated the glandular form of inflammation induced in the dorsolateral prostatic lobes (DLP) in the hormonal milieu of the decreased androgen/estrogen ratio. The anti-inflammatory action was seen in the decreased number of acini containing intraluminal neutrophils. As signs of antiestrogenic action, fispemifene blocked estrogen-induced expression of PR and Fra2 in the acinar epithelium of the DLP, and it decreased prolactin concentration in serum and the relative weights of the seminal vesicles and pituitary glands. Because fispemifene exhibited both antiestrogenic and anti-inflammatory action in the prostate, this experimental study suggests that SERMs could be considered as a new therapeutic option in the treatment and prevention of prostatic inflammation.

FISPEMIFENE, A NOVEL SERM, ATTENUATES GLANDULAR INFLAMMATION IN AN ANIMAL MODEL OF CHRONIC NONBACTERIAL PROSTATITIS

The anti-inflammatory and antiestrogenic action of fispemifene [Z-2-{2-[4-(4-chloro-1,2-diphenylbut-1-enyl)phenoxy]ethoxy}-ethanol], a novel selective estrogen receptor modulator (SERM), was tested on the Noble rat model of chronic nonbacterial prostatic inflammation with cellular composition and inflammation patterns similar to those described in human prostatitis. Inflammation was assessed by counting perivascular and stromal infiltrates and the number of inflamed acini. Furthermore, the aggressiveness of inflammation was assessed on the basis of the relation of lymphocytes to the acinar epithelium. The immunohistochemical expression of progesterone receptor (PR) and Fos-related antigen 2 (Fra2), prolactin concentration in serum, and the weights of the seminal vesicles and pituitary glands were used as endpoints of estrogen action. Fispemifene significantly attenuated the glandular form of inflammation induced in the dorsolateral prostatic lobes (DLP) in the hormonal milieu of the decreased androgen/estrogen ratio. The anti-inflammatory action was seen in the decreased number of acini containing intraluminal neutrophils. As signs of antiestrogenic action, fispemifene blocked estrogen-induced expression of PR and Fra2 in the acinar epithelium of the DLP, and it decreased prolactin concentration in serum and the relative weights of the seminal vesicles and pituitary glands. Because fispemifene exhibited both antiestrogenic and anti-inflammatory action in the prostate, this experimental study suggests that SERMs could be considered as a new therapeutic option in the treatment and prevention of prostatic inflammation.

Intravesical treatment with cis-urocanic acid improves bladder function in rat model of acute bladder inflammation

The aim was to study the effect of intravesically instilled cis‐urocanic acid (cis‐UCA) on bladder function in an experimental rat model of acute bladder inflammation. Hyaluronic acid (HA) was used as a comparator compound.

Galactoglucomannan-rich hemicellulose extract from Norway spruce (Picea abies) exerts beneficial effects on chronic prostatic inflammation and lower urinary tract symptoms in vivo

Galactoglucomannan (GGM) is the main hemicellulose class in wood of coniferous trees and could be potentially utilized as a possible health-promoting substance for food and pharmaceutical industry. Our aim was to evaluate effects of orally administered GGM-rich extract from Norway spruce in a rat model of chronic prostatitis associated with lower urinary tract symptoms (LUTS). Prostatic inflammation and LUTS was induced in male rats using testosterone and 17β-estradiol exposure for 18 weeks. Rats were treated with 2% GGM dissolved in drinking water during weeks 13-18. Pelvic pain response, LUT function and histopathological evaluation of the prostate were assessed. The results show that hormonal exposure induced LUTS seen as decreased urine flow rate, increased bladder pressure, voiding times, bladder capacity and residual urine volumes. GGM had positive effects on urodynamical parameters by decreasing the basal bladder pressure, increasing the urine flow rate and volume, reducing the residual volume and increasing micturition intervals. GGM reduced the extent of the hormone exposure-induced prostatic inflammation. Increase of pelvic pain induced by hormone exposure was only slightly affected by GGM treatment. The results suggest that orally administered GGM may have potential usage for improving lower urinary tract function associated with chronic prostatic inflammation.

Abstract 4968: Importance of tumor microenvironment in the preclinical estrogen receptor positive breast cancer- Primary tumor and bone metastasis models

Estrogen receptor positive (ER+) breast cancer has ability to metastasize to bone in high frequency. Bone is known to be fertile soil for metastasized cancer cells to survive and in turn, metastasized tumor cells alter normally balanced bone environment. Prevention and treatment of bone metastasis is challenging and better understanding why bone metastasis are resistant to current therapies is needed. Aim of the present study was to explore the role of tumor microenvironment and estrogen supplementation on growth of primary breast cancer tumor and bone metastasis to establish predictive ER+ breast cancer models for drug development. ER+ human breast cancer MCF-7 cells were inoculated into mammary fat pad and in the tibia of female athymic nude mice. Mice received either hormonal supplementation (17-beta estradiol pellet, E2) or placebo. Tumor growth was followed for 5 and 9 weeks. From one group, E2 supplementation was removed on study week 5 and tumor growth was followed for 4 weeks. During the study, blood samples were collected for serum steroid concentration measurements and at the end, histopathological evaluation and immunohistochemical (IHC) stainings were performed to examine tumor steroid hormone receptor expression. Orthotopic MCF-7 tumor growth was clearly hormone dependent. E2 supplementation, that increased serum estradiol for app. 3-fold, supported tumor growth and when E2 was removed, tumor size decreased and no tumor growth was observed thereafter. In the full absence of E2 supplement, no orthotopic tumor growth was observed. In contrast, when MCF-7 cells were inoculated into tibia, tumor growth was observed both with and without E2 supplement. IHC stainings confirmed orthotopic tumor to express ER, PR and AR when animals received E2. After E2 removal, orthotopic tumors expressed still ER and AR but no longer PR. Also intratibial tumors expressed ER and PR in the presence of E2, but no PR in the absence of E2 supplement. E2 is needed to support MCF-7 tumor growth when cancer cells are inoculated orthotopically into mammary fat pad. No orthotopic tumor growth is observed in the absence of E2 supplement. In contrast, in the bone microenvironment, MCF-7 cells form tumor even in the absence of E2 supplement. Results highlight importance of tumor microenvironment in the breast cancer progression and also refer why tumor in the different sites may be resistant to therapy. Taking together, when developing new therapies against breast cancer, focus should be addressed not only on primary tumor growth but also on bone metastasis where cancer cells are under influence of bone environment. Citation Format: Jenni Bernoulli, Mari I. Suominen, Tiina Kahkonen, Jenni Maki-Jouppila, Jussi M. Halleen, Riikka Oksala. Importance of tumor microenvironment in the preclinical estrogen receptor positive breast cancer- Primary tumor and bone metastasis models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4968. doi:10.1158/1538-7445.AM2017-4968

Activation of NF‐κB in Association with Prostate Carcinogenesis in Noble Rats

Abstract:  There is an increasing interest in the role of chronic nonbacterial prostatitis in the development of prostate cancer. The aim of the study was to explore the role of NF‐κB in the prostate of Noble rats treated with testosterone (T) and 17β‐estradiol (E2), a widely used model for prostate carcinogenesis. NF‐κB‐positive epithelial cells appeared in both inflamed and noninflamed glands and ducts at 13 weeks after hormone implantation in hypoandrogenemic, hyperestrogenemic rats. Both nuclear and cytoplasmic staining were observed. When daily dose of T was increased to give serum concentration above the level of control animals, dysplastic lesions and ductal carcinomas with NF‐κB‐positive cells were induced at 13 weeks and 26 weeks. The number of acini with NF‐κB‐positive cells decreased and no nuclear staining was observed. Surprisingly, no inflammation was seen in the periurethral region where ductal carcinomas developed. In conclusion, no unequivocal evidence was obtained to support the idea that NF‐κB would be activated in association with inflammation in the development of ductal carcinomas. The hormonal control of NF‐κB in the prostate warrants further studies.