Expression of estrogen receptor (ER)-alpha and ER-beta in normal and malignant prostatic epithelial cells: regulation by methylation and involvement in growth regulation

The aim of the current study is to demonstrate normal and malignant prostatic epithelial cells (PrECs) as targets for receptor-mediated estrogenic and antiestrogenic action. Using an improved protocol, we have successfully isolated and maintained highly enriched populations of normal PrECs from ultrasound-guided peripheral zone biopsies, individually determined to be morphologically normal. Semiquantitative reverse transcription-PCR analyses were used to determine whether transcripts of estrogen receptor (ER)-alpha and those of ER-beta were expressed in our normal PrEC primary cultures, in a commercially available PrEC preparation (PrEC; Clontech), in an immortalized PrEC line established from a benign prostatic hyperplasia specimen (BPH-1), and in three prostatic cancer cell lines (LNCaP, PC-3, and DU145). Expression levels of ER-alpha and ER-beta transcripts were related to those of two estrogen-responsive genes [progesterone receptor (PR) and pS2], at the message levels, to gain insights into the functionality of the ER subtypes in PrECs. Interestingly, only transcripts of ER-beta, but not those of ER-alpha, were found in our primary cultures of normal PrECs, along with both PR and pS2 mRNA. These data strongly suggest that estrogen action was signaled exclusively via ER-beta in normal human PrECs. In contrast, PrEC (Clontech) and BPH-1 cells expressed both ER-alpha and ER-beta transcripts and no PR nor pS2 mRNA in PrEC and only a minimal level of PR mRNA in BPH-1. Among the three prostate cancer cell lines, LNCaP expressed ER-beta mRNA along with transcripts of PR and pS2, DU145 expressed messages of ER-beta and PR, and PC-3 cells exhibited ER-alpha, ER-beta, and pS2 mRNA. Thus, unlike normal PrECs, expression patterns of these genes in malignant PrECs are more variable. Treatment of prostate cancer cells with demethylation agents effectively reactivated the expression of ER-alpha mRNA in LNCaP and DU145 and that of pS2 message in DU145. These findings provide experimental evidence that ER-alpha gene silencing in prostate cancer cells, and perhaps also in normal PrECs, are caused by DNA hypermethylation. To evaluate the potential of using antiestrogens as prostate cancer therapies, we have assessed the growth-inhibitory action of estrogens (estradiol and diethylstilbestrol) and antiestrogens (4-hydroxy-tamoxifen and ICI-182,780) on PC-3 and DU-145 cells. In PC-3 cells, which express both ER subtypes, estrogens as well as antiestrogens are effective inhibitors. In contrast, in DU145 cells, which express only ER-beta, antiestrogens, but not estrogens, are growth inhibitors. By comparison, ICI 182,780 is the more effective cell growth inhibitor. Importantly, the ICI 182,780-induced antiproliferative effects were reversed by cotreatment of DU145 cells with an ER-beta antisense oligonucleotide, hence lending additional support to a central role played by ER-beta in mediating growth-inhibitory action of antiestrogens

Charting the endometrial cancer care pathway : a baseline audit

Introduction: Longer waiting times from diagnosis to surgical resection have been found to negatively impact the overall survival and quality of life of women with endometrial cancer. The Cancer Care Pathway Directorate adopted the UK National Waiting Times Monitoring Dataset Guidance, to improve the timeliness of services along the cancer care pathway. From this, three key targets were identified: 1) Maximum 14-day wait from urgent GP referral for suspected cancer to first outpatient attendance (operational standard of 93%), 2) Maximum 31-day wait from decision to treat to first definitive treatment (operational standard of 96%), and 3) Maximum 62-day wait from urgent GP referral for suspected cancer to first treatment (operational standard of 85%). The aim of this baseline audit was to chart the time-frames of the various stages in the endometrial cancer pathway of patients diagnosed with this disease between 2015 and 2016 to assess for and identify delays in referral, investigation and care. -- Methods: A tool was developed following consultation with key stakeholders. Data protection clearance was obtained. Patient medical and oncology files, hospital databases, and MDT documentation for confirmed endometrial cancer cases were reviewed between September 2017 – March 2018. -- Results: A total of 101 endometrial cancer cases were included in the audit. The proportion of cases which met the 14-day, 31-day and 62-day wait KPIs operational standards were 39.1%, 81.2% and 17.2% respectively. -- Conclusion: The endometrial cancer care pathway timeframes did not meet the KPIs operational standards. Fast-track coordinators and nurse navigators could improve continuity and coordination of patient care.peer-reviewe

Pseudomonas aeruginosa Acute Prostatitis and Urosepsis after Sexual Relations in a Hot Tub▿

We report a case of a previously healthy 38-year-old male with acute prostatitis and concurrent Pseudomonas aeruginosa urosepsis. Pulsed-field gel electrophoresis analysis confirmed that the source of the organism was the patient's newly purchased hot tub, which was filled with water from a stream