Mutation analysis of the p51 gene and correlation between p53, p73, and p51 expressions in prostatic carcinoma.

BACKGROUND: p73 and p51 are genes possessing amino-acid similarities to p53. We previously found no mutation in p73 in prostatic carcinoma, but did find abnormal expression of the gene. Involvement of these genes in prostatic carcinogenesis is still poorly understood. METHODS: Mutation analysis of the p51 gene and allelotyping of 3q28, on which p51 lies, were performed. Expression of p53, p73, and p51 was examined using reverse transcription-polymerase chain reaction, and expression levels were compared. RESULTS: No mutation in p51 was found (0/55 cases). Loss of heterozygosity at 3q28 was detected in 6 of 28 cases (21.8%). By expression analysis we found that in p53, 4 of 38 cases (10.5%) showed downregulation. No cases showed upregulation of p53. In contrast, p73 and p51 were downregulated in 42.1 and 39.5% of cases, respectively, and upregulated in 31.5 and 34.2% of cases, respectively. Expression levels of p51 corresponded with those of p73 in 25 of 38 cases (65.8%). CONCLUSIONS: Somatic mutations in p73 and p51 are not important in prostatic carcinogenesis. These genes may be associated with tumors by expression levels and may have roles in addition to tumor suppression.

Prognostic status of p53 gene mutation in canine mammary carcinoma.

BACKGROUND: The p53 gene mutations have been associated with the development of human breast and canine mammary neoplasms; breast carcinoma patients with alterations of p53 gene are considered to have a poor prognosis. Mammary carcinoma represents the most common malignant tumor in female dogs. However, the prognostic significance of p53 gene mutation in the dog has been unclear. STUDY DESIGN: The alteration in exons 5-8 of p53 gene in 69 canine mammary carcinomas were investigated by PCR-SSCP with direct sequence analysis and statistically analyzed to compare with other clinicopathological parameters including age, neuter, tumor size, stage, histology, p53 expression, recurrence and death from carcinoma. RESULTS: 12 out of 69 (17%) carcinomas showed p53 gene mutations. After a follow-up period of 30 months, multivariate regression analysis revealed that p53 gene mutation was only an independent risk factor for increased risk of the recurrence and death from mammary carcinoma. CONCLUSION: The p53 gene alterations might contribute to the prognostic status in canine mammary carcinomas, in a way comparable to that of human tumors.

p53 gene mutations occurring in spontaneous benign and malignant mammary tumors of the dog.

Sixty-three cases of benign and malignant canine mammary tumors were analyzed to define the alteration of exons 5-8 for the p53 tumor suppressor gene using polymerase chain reaction direct sequence analysis with paraffin-embedded tissues. Four missense mutations were found in 38 benign mammary tumors (11%), and five missense (one tumor had two missense mutations) and one nonsense mutations were found in 25 mammary carcinomas (20%). These data suggest that the p53 gene alterations might be initiated at an early stage of canine mammary carcinogenesis and p53 mutations might be associated with malignancy. However, there was no evidence of any relationship between the p53 alterations and the histologic types of tumors or breeds of dogs.

Malignant phyllodes tumor of the prostate.

We report a case of malignant phyllodes tumor of the prostate which is the eleventh reported case in the world. Phyllodes tumor of the prostate is extremely rare and histologically resembles mammary phyllodes tumor. Phyllodes tumor of the prostate is classified into benign, borderline and malignant, but health professionals should carefully follow up the borderline cases in case they take a malignant clinical course. This case was the first to be treated by pre- and postoperative radiation therapy. Although the patient had a slight response to radiation therapy, he eventually developed metastasis. Because malignant phyllodes tumor of the prostate is a very aggressive tumor, people with the condition should undergo systemic chemotherapy as adjuvant therapy.

[Histopathological effect and its predictors of neoadjuvant hormonal therapy by combined androgen blockade].

OBJECTIVES: Combined androgen blockade (CAB) uning LH-RH agonist and flutamide has been performed as neoadjuvant therapy for T 2, 3 prostate cancers (CaP). The histological effects of neoadjuvant CAB therapy and influential factors were investigated. METHODS: Materials were 20 CaP cases which were underwent radical prostatectomy (RP) after neoadjuvant CAB therapy. All cases were diagnosed by echo-guided sextant needle biopsies. RP was performed after serum PSA was decreased to undetectable level. Histological effect was evaluated by general rule for clinical and pathological studies on prostate cancer (Japanese Urological Association). All cases were divided 2 groups by histological effects as follows: Group A (poor effect group): G 0 and G 1, Group B (good effect group): G 2 and G 3. Immunostaining of p 53 (mutant type), bcl-2 and Chromogranin A (ChA) were performed for both pretreatment needle biopsy and RP specimen. In addition, pretreatment serum PSA and Gleason grade were also investigated. RESULTS: Down grading were found in 30%. Down staging were found in 35% (7 cases). All 7 cases were negative surgical margins and 5 of 7 were clinical T 3. Negative bcl-2 of biopsy specimen was correlation with down grading (p = 0.008). In the histopathological evaluation, G 0 was 1, G 1 were 10, G 2 were 6 and G 3 were 3 cases. Gleason 4 or 5 elements of biopsy were found in 9/11 cases in Group A but only 3/9 cases in Group B (p = 0.027). The bcl-2 positive cells of biopsy were found in 8/11 cases in Group A but only 1/9 cases in Group B (p = 0.006). The p 53 and/or bcl-2 positive cells of biopsy were found in 10/11 cases in Group A but only 3/9 cases in Group B (p = 0.007). Serum PSA and ChA were not correlation with histological effect of neoadjuvant CAB therapy. But, in 3 cases, ChA positive cell appeared after neoadjuvant therapy. CONCLUSIONS: We could not expect more than 50% cases to show the down grading and down staging. But, in T 3 case, surgical failure could be decrease. We could expect prostate cancer cases without positive bcl-2 cells, p 53 over expression and Gleason 4 x 5 to reveal the good histological effects of neoadjuvant CAB therapy.

[Clinical studies on inverted papilloma of the urinary tract].

BACKGROUND: Inverted papilloma of the urinary tract is believed to be a benign neoplasm based on its histologic morphology and clinical behavior. In recent years, however, several investigators have warned against too optimistic an approach, emphasizing the possibility of malignant cellular transformation within the lesion, or its eventual association with other urothelial tumors such as transitional cell carcinoma or carcinoma in situ. We here report on 35 clinicopathologically diagnosed cases of inverted papilloma, and present the clinical significance attributed to these lesions in view of the current literature. PATIENTS AND METHODS: From 1976 to 1997, 35 cases of inverted papilloma of the urinary tract were treated at our hospital. This report presents the clinical features of these cases, the results of prognosis research, an investigation of the cases in which inverted papilloma and transitional cell carcinoma were found to co-exist, and a discussion of the recurrent cases of inverted papilloma found in previous literature. RESULTS: The patients ranged from 24 to 77 years of age, with a mean of 54 years, and included 4 women and 31 men. The most frequently occurring symptom was grosshematuria, and more than 90% of the 35 cases occurred in the bladder. In 2 of the 35 cases, transitional cell carcinoma coexisted with the inverted papilloma, at a different location in the bladder in one case and within the same neoplasm in the ureter in the other case. Clinical courses after treatment were followed in 29 of the 35 cases, with a follow-up period of from 8 months to 19 years (mean follow-up, 5 years and 4 months). Of these 29 cases, 2 showed recurrence, one at 16 and one at 30 months after the initial resection. Many previous reports show that the association of inverted papilloma and transitional cell carcinoma is stronger in the upper urinary tract and recurrence of inverted papilloma almost always happens with 2 years. CONCLUSION: This study suggests that some cases of urinary inverted papilloma show recurrence or malignant potential. Our results indicate that all cases of urinary inverted papilloma should be treated and followed as cases of low-grade transitional cell carcinomas. Consequently, all cases must be followed for two years or more after the initial operation.

Extragonadal germ cell tumor of the prostate associated with Klinefelter's syndrome.

PURPOSE: We report on a case of extragonadal germ cell tumor of the prostate associated with Klinefelter's syndrome. METHODS/RESULTS: The patient was a 33-year-old man. A transrectal prostate biopsy suggested combined germ cell tumor (yolk sac tumor + teratoma). Because there was no tumor except from the prostate, we considered this case to be a primary extragonadal germ cell tumor of the prostate. The prostate tumor responded to systemic chemotherapy with cisplatin, vinblastine and bleomycin and elevated lactate dehydrogenase and alpha-fetoprotein levels normalized. In addition to chemotherapy, the patient also underwent radiation therapy. CONCLUSION: The patient has survived for approximately 4 years since the diagnosis.

Expression of vascular endothelial growth factor in N-butyl-N-(4-hydroxybutyl)nitrosamine-induced rat bladder carcinogenesis.

Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are proteins implicated in tumor-associated microvascular angiogenesis. Expressions of VEGF and bFGF in various stages of chemical-induced rat bladder carcinogenesis were immunohistochemically investigated. Thirty-two male 6-week-old Wistar rats were given drinking water containing 0.05% N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) for 20 weeks. VEGF and bFGF were not detected in the normal bladder epithelium. In simple hyperplasia, intensive expression of VEGF was observed in a few epithelial cells, and the expression of epithelial VEGF became more pronounced in papillary or nodular (PN) hyperplasia and papilloma. In carcinoma, heterogeneous expression of VEGF was observed in focal tumor cells, intensely expressed in the invading tumor cells. Ultrastructurally, carcinoma cells showed VEGF immunoreactivity in the cytoplasmic matrix and some rough endoplasmic reticulum, and VEGF-positive and -negative carcinoma cells were also clearly defined. High levels of VEGF mRNA were observed in the carcinoma. However, bFGF was not detected in the epithelium throughout the carcinogenesis. Increased microvessel counts appeared at simple hyperplasia and became more pronounced in PN hyperplasia, papilloma, and carcinoma (F-test; P < 0.05). In the carcinoma, the microvessel counts of the VEGF-expressing tumor areas were significantly higher than that of the non-VEGF-expressing tumor areas (U-test; P < 0.05). The present study suggests that upregulation of epithelial VEGF may begin at a quite early stage in BBN-induced rat bladder carcinogenesis, but bFGF may not be involved.

Establishment and characterization of human choriocarcinoma cell line derived from a metastatic focus of a testicular mixed germ cell tumor.

A human testicular choriocarcinoma cell line HKRT-II was established by the single-cell cloning method from a mixed cell culture system derived from a retroperitoneal metastatic germ cell tumor composed of a yolk-sac tumor, a choriocarcinoma, and an immature teratoma. Its primary tumor rose from the testis and was comprised of a seminoma, a yolk-sac tumor, a choriocarcinoma and an immature teratoma. The HKRT-II cells were spindle or polygonal in shape and contained multi-nucleated giant cells showing neoplasticity and pleomorphism. The cells proliferated in a stable manner, and the population doubling time was 42 hours. The chromosome numbers showed a wide distribution of aneuploidy, while the mode was in the hypertetraploid range. Double minute chromosomes and homogeneously staining regions were recognized in about 5% to 10% of the metaphase plates, respectively. Heterotransplantation was not difficult. Subcutaneous transplantation of 1 x 10(7) cells into nude mice formed a tumor composed of only a choriocarcinoma. The most noteworthy characteristics of the cell line were that it produced human chorionic gonadotropin (hCG) in an in vitro culture system and in in vivo grafted cells, and that the N-myc gene was amplified about 10 times.

Cystic renal cell carcinoma: arose as a simple cyst, recurred repeatedly as a multicystic mass and finally presented as a solid mass.

An unusual case of cystic renal cell carcinoma in a 70-year-old Japanese male is reported. He had had a simple renal cyst removed 1 year ago. On presentation the right kidney was surrounded by multiple translucent cysts, which varied in size from 1 to 50 mm. The cyst walls were lined by single-layered cuboidal epithelium. The differential diagnosis included cystic mesothelioma, cystic lymphangioma and multicystic dysplastic kidney. An immunohistochemical and ultrastructural study and flow cytometric analysis of DNA ploidy were performed. The tumor was differentiated to such an extent that it was difficult to diagnose as carcinoma; however, it recurred repeatedly. Three and a half years after initial presentation the tumor had invaded the ileum with the pathological change to be almost solid when viewed grossly and, microscopically, showed tubulo-papillary structures in addition to a cystic pattern. The DNA ploidy pattern revealed a near-diploid aneuploid in the early specimen and a polyploid aneuploid in the last specimen.

Mutation, allelotyping, and transcription analyses of the p73 gene in prostatic carcinoma.

A novel gene, p73, encoding a protein with significant homology to p53, was recently identified at 1p36. To investigate penetrance of p73 in prostatic carcinogenesis, mutation, allelotyping, and transcription analyses of p73 were performed in prostatic carcinoma. No types of mutation causing amino acid substitutions or frameshifts were found in 106 cases examined. Loss of heterozygosity in the gene was found in 2 of 38 cases (5.3%). Various expression levels of p73 alpha variant were observed in tumor compared with those in normal tissue. These data suggest that the p73 gene is not playing an essential role, but expression of p73 may associate with tumor growth in prostatic carcinogenesis.

[Prostatic involvement of bladder carcinoma].

BACKGROUND: The histological pattern of prostatic involvement by transitional cell carcinoma is still unclear. The present study was carried out in bladder carcinoma with prostatic involvement to clarify the histological invasion pattern and its association with primary lesions. METHODS: In the past 10 years, 83 cases of total cystectomy including prostatectomy underwent pathological diagnosis in our department. This study included 81 cases of transitional cell carcinoma (TCC), of which 11 showed prostatic involvement of bladder carcinoma. In these cases, the histological patterns of invasion were classified in relation to prostatic urethra, prostatic duct, stroma, lymphatic duct, capsule, vein and perineural invasion. Seventy cases without prostatic involvement were controls. The location, pathological grade, stage and lymphatic involvement of primary bladder carcinoma were compared in terms of prostatic involvement cases with control cases. RESULTS: Among those 11 cases, there were 3 cases in which only the prostatic duct was involved, 2 cases with invasion to only lymphatic duct, and involvement of both in 6 cases. One case of the prostatic duct involvement showed non-continuous invasion in the prostatic duct without prostatic urethra invasion, suggesting the possibility that non-continuous invasion could occur as a type of multicentric growth of TCC. CONCLUSIONS: Suspected routes of invasion of bladder carcinoma into the prostate were; 1 continuous transductal, 2 trans-lymphatic ductal, 3a combination of the two. It appears necessary to consider the possibility of TCC occurring in the prostate simultaneously with bladder carcinoma as a part of multicentric growth. There was a tendency of prostatic involvement cases include the bladder neck and trigone, and show lymphatic duct involvement more than non-prostatic involvement cases.

The behavior of endometrial hyperplasia: a prospective study. Endometrial Hyperplasia Study Group.

OBJECTIVE: To clarify the behavior of endometrial hyperplasia in a prospective study. METHOD: Fifty-one patients with endometrial hyperplasia were followed up for 6 months. Samples of endometrial tissues were taken by uterine endometrial biopsy every 4 weeks during the first 3 months and at the end of follow-up. RESULTS: In 69% (35/51) of the patients histological picture of the endometrium became normal during the observation period. The lesions persisted in 17% (6/35) of the patients with simple hyperplasia, in 25% (1/4) of those with complex hyperplasia, in 14% (1/7) of those with simple atypical hyperplasia, and in 80% (4/5) of the patients with complex atypical hyperplasia. In the remaining 3 patients with simple hyperplasia, the lesions progressed to complex atypical hyperplasia by the end of follow-up, after showing a normal endometrium. CONCLUSION: Most cases of endometrial hyperplasia, except for complex atypical hyperplasia, disappeared spontaneously within a short period of time.

Testicular efferent ductule cyst of a dog.

A 9-year-old male Shetland Sheepdog had a small mass in the left testis. Grossly, the round to oval cyst was present at the upper pole of the testicular parenchyma near the head of the epididymis. Histologically, the cyst was lined by a single layer of nonciliated and ciliated epithelial cells. Immunohistochemically, the epithelial cells of the cyst showed expression of the low- and high-molecular-weight cytokeratins, vimentin, and desmin similar to that of normal efferent ductules in the dog. The testicular cystic dysplasia was thought to originate from the efferent ductules.

p16 (CDKN2/MTS1) gene deletions are rare in prostatic carcinomas in the United States and Japan.

PURPOSE: The incidence of clinically apparent prostatic carcinoma is much higher in the United States than in Japan. Alterations in the p16 tumor suppressor gene have been identified in various tumor types, including cultured prostatic carcinoma cell lines. We studied the possible deletions of either exon 2 or 3 of this gene in primary clinical prostatic carcinomas from Japan and the United States. MATERIALS AND METHODS: Genomic DNA was extracted from 36 formalin-fixed, paraffin-embedded clinical prostatic carcinomas from Japan and 27 carcinomas from the United States. Exons 2 and 3 of the p16 gene were amplified using comparative multiplex polymerase chain reactions (PCR) and then analyzed for possible deletions of either exon. RESULTS: Two out of 36 (5.6%) carcinomas from Japan clearly demonstrated deletion of p16 exon 2, but this deletion was not detected in any of the 27 carcinomas from the United States. CONCLUSIONS: Although slightly higher in Japan than in the United States, the frequency of p16 exon deletions in clinical prostatic carcinomas is very low, and probably is not important in the development of this neoplasm.

[Image cytometric DNA analysis in bladder tumors].

UNLABELLED: We studied image cytometric DNA analysis of bladder tumors to evaluate malignant potentials of bladder tumors. METHODS: Thirty nine samples were obtained by TUR from 37 patients. Nuclear DNA content of all samples were measured by image cytometer and were determined ploidy pattern by DNA histogram. RESULTS: Of 39 TCC non-diploid pattern was recognized in 50% of grade 1 cases, 73% of grade 2 cases and 100% of grade 3 cases. DNA ploidy was strictly correlated with histological grading in TCC. DNA non-diploid pattern was present in 67% of papillary tumors, 87.5% of non-papillary tumors and 100% in CIS. In diploid pattern 2 of 7 cases with grade 1 and 2 of 4 cases with grade 2 recurred. In non-diploid pattern 1 of 4 cases with grade 1, 4 of 10 cases with grade 2 and 4 of 6 cases with grade 3 recurred. There was no significant correlation between diploid and non-diploid pattern in grade 1, 2, 3. CONCLUSION: Image cytometric DNA analysis may be useful in addition to the classic and prognostic parameters of stage and grade, especially in TCC. The differences between image analysis system and flow cytometric analysis for DNA measurement were discussed.

Lectin histochemical evaluation of glycoconjugates in dog efferent ductules.

Glycoconjugates in the epithelial cells of the efferent ductules in the dog were investigated using lectin histochemistry. These ductules connect the extratesticular rete with the epididymis. The epithelium of the ductules consisted both of ciliated and nonciliated cells. Whereas the apical zone of ciliated cells showed selective binding with WGA, SWGA, SNA, MAA and neuraminidase-PNA, that of nonciliated cells bound to all lectins used in the present study: WGA, SWGA, SNA, MAA, PNA, neuraminidase-PNA, RCA1, DBA and SBA. The nonciliated cells were divided into 3 types: type A cells which lacked both specific granules and vacuoles, type B cells which were characterised by a few specific apical vacuoles and many large specific granules, and type C cells which were characterised by some specific apical vacuoles and small basal granules. The specific granules and vacuoles of type B cells showed binding with WGA, SWGA and MAA. The specific granules of type C cells showed binding with WGA, SWGA, SNA, MAA, PNA and neuraminidase-PNA, while their specific vacuoles showed binding with WGA, SWGA, SNA and MAA. The Golgi zone both of ciliated and type A cells did not bind with any lectins used in this study, while type B and C cells showed similar lectin binding patterns between the Golgi zone and their specific granules. Specific lectin binding patterns revealed a different carbohydrate composition of each type of cell, indicating a biological difference between the ciliated cells and the 3 types of nonciliated cells in dog efferent ductules.

Immunohistochemical localization of the epidermal growth factor-receptor in rhesus-monkey prostate.

Epidermal growth-factor receptor (EGF-r), a membrane-bound glycoprotein activated by EGF, is important in maintaining the integrity and function of the prostate. To investigate EGF-r presence in the prostate of the rhesus monkey, monoclonal-antibody immunohistochemical examination was performed. The monkey prostate consisted of the cranial and caudal lobes, and the prostatic epithelial cells were composed of the secretory and basal cells. The distribution patterns of EGF-r in the prostatic epithelial cells were quite different between the cranial and caudal lobes. In the caudal lobe, EGF-r was seen in both secretory and basal cells, whereas, in the cranial lobe, the EGF-r was seen exclusively in the basal cells. The stromal cells of both lobes did not show EGF-r. This study revealed that each prostatic lobe contains specific binding sites for EGF, indicating a biological difference between the two lobes of the prostate of the rhesus monkey.