Androgen receptor mutation (T877A) promotes prostate cancer cell growth and cell survival.

Alteration of the AR functions due to amplification, overexpression and somatic mutation of the AR itself or altered interaction of AR with other cell growth regulatory proteins, may contribute to a significant subset of advanced prostate cancer (CaP). Very little is known about the pathways impacted by AR dysfunctions, although CaP associated AR alterations suggest the biological role of the AR dysfunction in disease progression. Comparative evaluations of wild type (wt) AR and mutant (mt) ARs in appropriate experimental models should provide a better understanding of the functional impact of AR alterations in CaP. Here, we provide direct evidence showing cell growth/cell survival promoting effects of the widely studied CaP associated AR mutation (T877A). In contrast to Ad-wtAR or Ad-control infected LNCaP or LAPC4 cells, Ad-mtAR (T877A) infected LNCaP or LAPC4 cells continued to grow in the androgen-deprived medium and exhibited an androgen independent AR-transcription factor activity. Further, Ad-mtAR (T877A) infected LNCaP or LAPC4 cells exhibited enhanced cell growth in the presence of lower concentrations of the synthetic androgen, R1881. Of note, Ad-mtAR (T877A) infected LNCaP cells showed striking resistance to cell growth inhibition/apoptosis mediated by the wt p53. Taken together, these findings provide novel insights into the AR dysfunctions resulting from the T877A mutation and functionally similar AR alterations may provide selective cell growth/survival advantage for CaP progression. These observations have important implications for developing biology-based prognostic biomarkers and therapeutic strategies for CaP showing such AR dysfunctions.

Quantitative expression profile of PSGR in prostate cancer.

PSGR is a novel member of the G-protein-coupled olfactory receptor family. Our initial report showed predominant expression of the PSGR in human prostate gland and significant alterations of PSGR expression in primary prostate cancer (CaP) specimens. The aim of this study was to provide in-depth evaluations of the expression profile of PSGR in prostatic epithelial cells of CaP patients and to evaluate the association of PSGR expression characteristics with clinico-pathologic features. In total, 220 RNA specimens, from laser capture microdissected paired benign and malignant prostatic epithelial cells of 110 CaP patients, were analyzed for PSGR expression by quantitative real-time PCR. The differential expression of PSGR between the prostatic epithelial cells of malignant and benign glands was statistically significant (P<0.0001). Comparison of PSGR expression between paired benign and tumor cells revealed prostate tumor cell-specific overexpression in 67.2% of tumor specimens (74 of 110), decreased expression in 20.9% of tumor specimens (23 of 110) and no difference of PSGR expression between tumor and normal cells in 11.8% of specimens (13 of 110). In representative cases, PSGR expression patterns were independently confirmed by in situ RNA hybridization. The PSGR overexpression associated with higher percentage of pathologic stage, pT3, and a higher level of preoperative serum PSA. CaP cells of African-American CaP patients exhibited about two-fold increase of PSGR expression in comparison to the Caucasian American CaP patients. Strikingly high-percentage CaP cells overexpress PSGR warrants further studies of PSGR expression alterations to define subsets of CaPs.

Frequent detection of codon 877 mutation in the androgen receptor gene in advanced prostate cancers.

Prostatic tissue specimens derived from transurethral resections of patients with metastatic prostate cancer were analyzed for genetic alterations in the hormone-binding domain of the androgen receptor (AR) gene. Direct sequencing of the polymerase chain reaction-derived DNAs of 6 of 24 specimens revealed a codon 877 mutation (ACT-->GCT, Thr-->Ala) in the hormone-binding domain of the AR gene. This same AR mutation has been reported previously in a metastatic prostate cancer cell line, LNCaP, where this mutation confers upon the AR an altered ligand-binding specificity which is stimulated by estrogens, progestagens, and antiandrogens. It is possible that analogous to an activated/altered growth factor receptor oncogene, codon 877 mutant AR with altered ligand binding may provide a selective growth advantage in the genesis of a subset of advanced prostate cancer. Although estrogens are used infrequently, antiandrogens are used increasingly in hormonal therapy for patients with advanced prostate cancer. The stimulatory effect of these therapeutic agents on the codon 877 mutant AR further suggests that this frequently observed AR mutation may contribute to the treatment refractory disease.

PSGR, a novel prostate-specific gene with homology to a G protein-coupled receptor, is overexpressed in prostate cancer.

PSGR, a new prostate tissue-specific gene with homology to the G protein-coupled odorant receptor gene family, has been identified. Here we report the characteristics of the predicted protein sequence of PSGR and its prostate tissue specificity and expression profile in human prostate cancer and matched normal tissues. Using multiple tissue Northern blots from over 50 different tissues, PSGR expression was restricted to human prostate tissues. Paired normal and tumor specimens from 52 primary prostate cancers, obtained by laser capture microdissection or manual microdissection, were analyzed for PSGR expression by semiquantitative and real-time PCR assays. The differential expression of PSGR between normal and tumor tissues was highly significant (P < 0.001), and 32 of 52 (62%) matched prostate specimens exhibited tumor-associated overexpression of PSGR. Of note, there was very little or no expression of PSGR in many normal specimens in comparison with the generally high expression of PSGR seen in matched tumor specimens. In situ hybridization assays showed restricted PSGR expression in the epithelial cells of the normal and tumor tissue sections. Restricted expression of PSGR in prostatic epithelial cells, overexpression of the PSGR in a significant percentage of prostate cancers, and the predicted protein sequence of PSGR with seven transmembrane domains provide a foundation for future studies evaluating the potential of PSGR as a prostate cancer gene expression marker and the utility of PSGR protein as a novel target for developing immunotherapeutic strategies for prostate cancer.

A novel human cancer culture model for the study of prostate cancer.

Research into molecular and genetic mechanisms underlying prostate carcinogenesis would be greatly advanced by in vitro models of prostate tumors representing primary tumors. We have successfully established an immortalized human prostate epithelial (HPE) cell culture derived from a primary tumor with telomerase. The actively proliferating early passaged RC-58T cells were transduced through infection with a retrovirus vector expressing the human telomerase catalytic subunit (hTERT). A high level of telomerase was detected in RC-58T/hTERT cells but not RC-58T cells. RC-58T/hTERT cells are currently growing well at passage 50, whereas RC-58T cells senesced at passage 7. RC-58T/hTERT cells exhibit transformed morphology. More importantly, these immortalized cells showed anchorage-independent growth as they formed colonies in soft agar and grew above the agar layer. Expression of androgen-regulated prostate specific gene NKX3.1 and epithelial specific cytokeratin 8 (CK8) but not prostate specific antigen (PSA) and androgen receptor was detected in RC-58T/hTERT cells. Prostate stem cell antigen (PSCA) and p16 were also expressed in this cell line. RC-58T/hTERT cells showed growth inhibition when exposed to retinoic acid and transforming growth factor (TGF)-beta1 known potent inhibitors of prostate epithelial cell growth. A number of chromosome alterations were observed including the loss of chromosomes Y, 3p, 10p, 17p, 18q and the gain of chromosomes 16 and 20. These results demonstrate that this primary tumor-derived HPE cell line retained its transformed phenotypes and should allow studies to elucidate molecular and genetic alterations involved in prostate cancer. This is the first documented case of an established human prostate cancer cell line from a primary tumor of a prostate cancer patient with telomerase.

p53 nuclear protein expression is an independent prognostic marker in clinically localized prostate cancer patients undergoing radical prostatectomy.

Immunohistochemical (IHC) staining for p53 protein nuclear expression was evaluated in archival paraffin-embedded radical prostatectomy specimens from 139 patients with clinically localized prostate cancer followed up from 1 to 8 (mean, 4) years. Elevated nuclear p53 protein expression was detected in 85 (61%) of 139 patients, being heterogeneous and focal in the majority of specimens. Only four specimens displayed homogeneous nuclear accumulation of p53 protein. Disease progression, most commonly prostate-specific antigen elevation, was noted in 46 (33%) patients, with 39 (85%) having positive p53 protein IHC stains. Conversely, 93 (67%) of 139 have not recurred, with 46 (49%) having positive p53. Of all 54 p53-negative patients, 47 (87%) have had no disease recurrence. An increased p53 protein IHC stain was associated with a higher pathological stage (T1 and T2, 51% versus >/=T3, 69%) and Gleason score 2-4, 17%; 5-7, 72%; and 8-10, 87.5%). Despite these associations, p53 IHC staining was an independent predictor of disease-free survival in a multivariate analysis of p53, age, race, stage, and grade. This study revealed that a majority of clinically localized prostate cancers heterogeneously express elevated nuclear levels of p53 protein in at least a subset of malignant cells, and that this expression is an independent predictor of disease progression in prostate cancer patients after radical prostatectomy.

Phenology and field biology of black cutworm (Lepidoptera: Noctuidae) in Ontario no-till corn.

Black cutworm, Agrotis ipsilon (Hufnagel), is an occasional corn, Zea mays L., pest that is attracted to no-till fields. Understanding the phenology of black cutworm in Ontario no-till corn, particularly the time of arrival of adults in relation to the onset of crop damage and the stages of larvae that coincide with vulnerable corn seedling leaf stages, is important for their effective control. Pheromone and blacklight trap captures of moths first occurred in early April, whereas significant influxes did not occur until mid- to late April. Males and females were often captured simultaneously, in contrast to findings in Iowa and Illinois where males were captured in pheromone traps on average 3 wk ahead of females or males in blacklight traps. This may be a reflection of a more mature source population for the influxes into Ontario because first captures also were later than in the United States. Females arrived mated and corn seedling cutting occurred within 137 degree-days (DD) (base 10.4 degrees C) of first capture in Ontario corn. Cutworms were present in cornfields before planting, and the mean age of larvae increased along with corn leaf stage, suggesting that no new recruitment took place after planting. The apparent synchrony between corn and cutworm phenology in the northern areas of corn production seems more related to the availability and quality of food for young larvae relative to the development of the crop then the time of arrival of moths.

National Cancer Institute study of luteinizing hormone-releasing hormone plus flutamide versus luteinizing hormone-releasing hormone plus placebo.

A randomized, double-blind trial in patients with disseminated, previously untreated prostate cancer (stage D2) was designed to test the hypothesis that maximal androgen blockade improves the effectiveness of the treatment of prostatic cancer. Six hundred three men received leuprolide in combination with either placebo or flutamide, and were followed for a minimum of 5 years. The 303 patients randomly assigned to receive leuprolide and flutamide had a longer progression-free survival and an increase in the median length of survival compared with the 300 patients receiving leuprolide plus placebo. Differences between the treatments were particularly evident for men with minimal disease and good performance status.

Protein expression of p53, bcl-2, and KI-67 (MIB-1) as prognostic biomarkers in patients with surgically treated, clinically localized prostate cancer.

BACKGROUND: Protein expression in the primary tumor of the tumor suppressor gene p53 and the proto-oncogene bcl-2 have been shown to be prognostic biomarkers of cancer recurrence after radical prostatectomy in patients with clinically localized prostate cancer. Cancer cell proliferation as measured by immunohistochemical markers such as the MIB-1 antibody for Ki-67 has recently been suggested to be of prognostic value in prostate cancer. The goal of this study was to determine the clinical use of p53, Ki-67 (MIB-1), and bcl-2 immunohistochemical protein expression in the primary tumor as combined predictors of disease progression after radical prostatectomy (RP). METHODS: Protein expressions of p53, Ki-67, and bcl-2 were evaluated in archival paraffin-embedded RP specimens from 162 patients monitored from 1 to 10 years (mean, 4.5 years) and correlated to stage, grade, race, and serologic (prostate-specific antigen) recurrence after operation. RESULTS: Expression was detected in 112 (69.1%), 44 (27.2%), and 62 (38.3%) of 162 patients for p53 (1+ or greater), bcl-2 (1+ or greater), and Ki-67 (2+ or greater), respectively. Biomarker expressions were not correlated to age and race; however, all increased with increasing stage and grade. The degree of expression by percentage of malignant cells staining correlated to recurrence for p53 and Ki-67 but not for bcl-2. All three markers were correlated to raw and Kaplan-Meier recurrence by means of univariate analysis with recurrence estimates at 6 years of 60.7% versus 24.2%, 84.2% versus 38.6%, and 72.4% versus 30.6% comparing positive versus negative expression of p53, bcl-2, and Ki-67, respectively. p53 and bcl-2 remained as independent prognostic markers by Cox multivariate regression analysis. Although Ki-67 did not remain an independent marker, it added prognostic use in certain subsets of patients. CONCLUSIONS: p53, bcl-2, and Ki-67 (MIB-1) appear to be important biomarkers to predict recurrence in patients with clinically localized prostate cancer after RP, and all three biomarkers deserve further study.

Postoperative myocardial infarction after radical cystoprostatectomy masked by patient-controlled analgesia.

We present a case report where improper patient use of patient-controlled analgesia (PCA) in the postoperative period resulted in a significant delay in diagnosis of a postoperative myocardial infarction. Despite its excellent safety record and documented efficacy in controlling pain, PCA does have its limitations and can present a danger to the patient if operator error, patient error, or mechanical errors occur. Although the latter is rarely of concern, the two former possibilities exist. Other reported complications of PCA are discussed. We recommended that physicians, when considering use of patient-controlled anesthesia, discuss the qualitative and quantitative aspects of pain associated with the particular type of surgery performed to avoid missed postoperative complications.

Hepatic subcapsular extension of pelvic lymphocele after radical retropubic prostatectomy.

Lymphoceles are a rare symptomatic complication after radical prostatectomy occurring in less than 5% of cases. We present a case of a symptomatic lymphocele that occurred after a radical retropubic prostatectomy and obturator lymphadenectomy. The lymphocele dissected along the retroperitoneum and extended into the hepatic subcapsular space and became secondarily infected with Candida albicans.

Defining the role of antiandrogens in the treatment of prostate cancer.

OBJECTIVES: Although antiandrogens have been used as monotherapy and in combination with other treatment modalities for management of metastatic prostate cancer, their major role to date has been one in which they are used in conjunction with surgical or medical castration for treatment of Stage D (T4/Nx/M1) carcinoma of the prostate. The widespread use of prostate-specific antigen (PSA) is increasing the number of men who are diagnosed with earlier stages of this disease, thus resulting in a greater number of definitive therapeutic procedures. Also, PSA has become the primary modality for following these patients after definitive treatment. Because the use of PSA results in the discovery of lower volume of disease and in discerning earlier recurrence of disease, the question arises as to whether an antiandrogen alone could be an adequate treatment in both neoadjuvant and adjuvant settings. METHODS: As data accumulate that point to the efficacy of combined androgen blockade in metastatic disease, a protocol has been developed to test the hypothesis of intermittent combined treatment in patients who present with minimal disease and good performance status. In this study, the antiandrogen bicalutamide will be used in combination with the luteinizing hormone-releasing hormone (LHRH) analogue goserelin acetate. Also, short-term combined androgen ablation continues to be investigated prior to both radiation therapy and radical prostatectomy for localized disease. RESULTS: Two groups of patients are ideal candidates for the use of antiandrogen monotherapy following radical prostatectomy: those whose PSA values do not fall to undetectable levels and the far larger group of men who have capsular penetration or positive surgical margins with nondetectable PSA levels. Protocols have been developed to assess the clinical potential of bicalutamide as the sole adjuvant therapy in these postprostatectomy patients, based in no small part on the relative paucity of side effects of this antiandrogen. CONCLUSIONS: Antiandrogens in general, and bicalutamide in particular, are poised to play an increasing role in the treatment of all stages of adenocarcinoma of the prostate. The safety profile of bicalutamide, combined with its long half-life, resulting in once-a-day dosing, makes it an ideal choice for clinical trials evaluating an antiandrogen, either alone or combined with reversible medical castration, in earlier stages of prostate cancer as well as in various combination regimens in metastatic disease.

Low-velocity gunshot injury to ureter.

An unusual type of ureteral injury is described. The conventional surgical management of a penetrating ureteral injury was not utilized. Because debridement was not necessary in this case, we did not use an internal ureteral stent. A good postoperative result was obtained with this conservative treatment.

Comparison of Megace, Stilphostrol, Megace plus DES, or streptozotocin in metastatic prostatic cancer in patients with hormonal failure and prior radiotherapy.

The National Prostatic Cancer Project from 1982 to 1985 evaluated several treatments for metastatic prostatic cancer patients who had a history of prior radiotherapy and were refractory to hormone manipulation. The treatments studied were megestrol acetate (Megace), Megace plus diethylstilbestrol (DES), diethylstilbestrol diphosphate (Stilphostrol), and streptozotocin. While the four treatment arms did not differ significantly with respect to survival, there was a small but significant difference in progression-free survival among the treatment groups. These patients are difficult to treat and have many secondary problems, and perhaps future studies of current and new agents should focus more on subjective and other secondary benefits for them.

Efficacy of radiographic chest imaging in patients with testicular cancer.

OBJECTIVE: To determine the efficacy of computed tomography of the chest (CTC) and plain radiograph (CXR) in the initial staging process of testicular germ cell tumors. METHODS: The medical records of 362 patients with testicular germ cell tumor treated at our center between January 1980 and August 1993 were reviewed with particular attention to initial chest screening studies. Two hundred one patients had both CXR and CTC, 24 CXR alone, and 20 CTC alone during initial staging. One hundred seventeen patients were excluded from analyses because of undergoing whole lung tomography (92), unknown staging (19), or inadequate follow-up (6). Analysis included findings based on abdominal staging results using computed tomography of the abdomen (CTA). RESULTS: Of the 201 patients who had both CTC and CXR, 117 (58.2%) had nonseminomas (NSGCT) and 84 (41.8%) had seminomas (SEM). Among the patients with NSGCT, 21 (17.9%) had chest metastasis, 16 (76.2%) of which were detected by CXR. The 5 that were missed on CXR had significant retroperitoneal disease documented by CTA and the knowledge of chest metastases potentially altered therapy in 2 patients. Only 2 of 84 (2.4%) patients with SEM had metastatic chest disease and both were identified by CXR. False-positive CTC following negative CXR resulted in costly and sometimes invasive additional procedures in 10 patients with NSGCT and 6 with SEM. None of the CXR-only patients had adverse consequences from the solitary study (at least 1 year follow-up). The CTC-only patients could have undergone CXR only and had similar outcome. CONCLUSIONS: CXR alone is preferable for initial chest staging in all patients with SEM and in patients with NSGCT with negative findings on CTA. CTC remains of slight benefit for patients with clinical Stage II and greater NSGCT and to evaluate further suspicious CXR findings in any patient, although it appears not to be necessary in patients who have clinical Stage I disease determined by CTA. These findings have important cost-saving implications.

Adrenal cortical carcinoma with vena cava tumor thrombus requiring cardiopulmonary bypass for resection.

We believe this is the fifteenth case report of adrenal cortical carcinoma with tumor thrombus to the vena cava, and the fourth reported case of a left-side tumor propagating thrombus to the vena cava. The patient underwent successful resection which required cardiopulmonary bypass. The caval tumor thrombus was very friable and gelatinous, unlike many renal cell thrombi, and required special surgical considerations.

Critical appraisal of routine supraclavicular lymph node biopsy in staging of testicular tumors.

Supraclavicular lymph node biopsy was performed as a staging procedure in 36 patients with germ cell tumors of the testis and nonpalpable supraclavicular nodes. Of 28 patients with clinical Stage A or B disease, 1 patient (4 per cent) was found to have supraclavicular metastases. Of 8 patients with clinical Stage C disease, 2 (25 per cent) had supraclavicular metastases. The apparent infrequency with which subclinical supradiaphragmatic disease is documented with this procedure and the current use of adjuvant systemic therapy in patients with pathologic Stage B nonseminomatous tumors suggest that supraclavicular lymph node biopsy should be abandoned as a routine staging procedure.

Transitional cell carcinomatous meningitis after M-VAC (methotrexate, vinblastine, doxorubicin, and cisplatin) chemotherapy.

The M-VAC (methotrexate, vinblastine, doxorubicin, and cisplatin) regimen has been utilized at our two institutions to treat 17 patients with advanced stage transitional cell carcinoma of the bladder. We report 2 cases of carcinomatous meningitis resulting from metastatic transitional cell carcinoma which occurred in patients treated with M-VAC. Review of the literature suggests that our experience with central nervous system metastases is not unique, and that treatment of advanced stage transitional cell carcinoma of the bladder with M-VAC may enhance the incidence of meningeal metastases. Carcinomatous meningitis, although rare, is a rapidly fatal manifestation of metastatic transitional cell carcinoma if left untreated. However, prompt diagnosis and early aggressive therapy may result in palliation and stabilization of neurologic status. We review the pathophysiology, diagnosis, and treatment of transitional cell carcinomatous meningitis.

Estimating the cost effectiveness of total androgen blockade with flutamide in M1 prostate cancer.

OBJECTIVES: Although combined androgen blockade with flutamide plus medical or surgical castration is effective in metastatic prostate cancer, debate exists over whether it is cost effective. METHODS: Decision analysis model of hypothetical cohorts of 70-year-old men presenting with metastatic prostate cancer, using a societal perspective, calculated anticipated survival and incremental cost per life-year gained. Time to progression and survival rate were from the Intergroup 0036 trial. Costs were based on Medicare data and wholesale drug pricing. Flutamide was estimated to reduce the relative risk of progressive disease by 25% (range, 0 to 50%). Costs and survival benefits were discounted at a 5% annual rate. RESULTS: In our model for minimal disease, median survival increased from 42.3 to 49.4 months with flutamide and average survival by 5.2 months at an incremental cost of $25,300 per life-year gained. If the efficacy were as high as 50%, the benefit would be 12 months at a cost of $13,700 per life-year gained. At a 10% efficacy, the benefit would be 1.9 months at a cost of $60,900 per life-year gained. For severe disease, the model estimated the median survival increased from 29.5 to 34.3 months with flutamide and average survival by 4.0 months at an incremental cost of $20,000 per life-year gained. At worst-case 10% efficacy, the benefit decreased to 1.5 months at an incremental cost of $47,500 per life-year gained. Total costs for patients treated with an orchiectomy and flutamide compared to leuprolide alone were similar if severe disease was present and actually lowered costs if there was minimal disease. CONCLUSIONS: Flutamide has an incremental cost effectiveness more favorable than most accepted therapies. If drug costs are covered under health care reform, flutamide should be initiated and covered for all good performance status patients.

Clinical evolution of cystic teratoma during treatment with combination chemotherapy.

Progressive, cystic tumor enlargement in the abdomen developed in a patient with teratocarcinoma during treatment with systemic chemotherapy. Tumor markers were elevated in the cyst fluid and negative in serum. Further, the patient underwent a successful surgical debulking of large amounts of cystic teratoma.