Expression of angiogenesis-related genes regulates different steps in the process of tumor growth and metastasis in human urothelial cell carcinoma of the urinary bladder.

OBJECTIVE: This study was designed to determine the relative activity of angiogenesis-related genes in the regulation of tumorigenicity and subsequent metastases of urothelial cell carcinomas (UC) of the urinary bladder. METHODS: We selected the clones with the highest and lowest expression level of basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF)/vascular permeability factor or interleukin-8 (IL-8) in the highly tumorigenic and metastatic human UC cell line 253J B-V. Tumorigenicity and production of spontaneous lymph node metastases were evaluated 1, 2, 4, 8 and 12 weeks after orthotopic implantation of each specific expression clone into the urinary bladder of athymic nude mice. Moreover, the transitional changes in the expression of angiogenesis-related genes and neovascularization were determined in tumors and metastases. RESULTS: At the early stage of tumor growth following orthotopic implantation, tumorigenicity and metastases were significantly increased in the clones with the highest expression of bFGF and IL-8, while they were significantly inhibited in the clones with the lowest expression of bFGF and IL-8 compared to parental 253J B-V. In the tumors, specific expression of angiogenesis-related genes and intratumor neovascularity of each clone were gradually regulated to the same level as parental 253J B-V. In metastasized tumors of the highest and lowest IL-8-expressing clones, IL-8 expression was consistently high and low, respectively. CONCLUSIONS: These findings indicate that at the early stage of tumor growth, bFGF and IL-8 expression play important roles in the regulation of angiogenesis, tumorigenicity and subsequent metastases of human bladder cancer.

Detection of circulating cancer cells with von hippel-lindau gene mutation in peripheral blood of patients with renal cell carcinoma.

Mutations of the von Hippel-Lindau (VHL) tumor suppressor gene have been detected in up to 60% of sporadic clear cell renal carcinomas (RCCs). Even patients with RCCs believed to be curable with radical nephrectomy sometimes develop distant metastasis 5-10 years after surgery, suggesting hematogenous circulation of cancer cells. Useful tumor markers have not yet been established for RCC. To detect patients at high risk of metastasis after surgery, we developed a highly sensitive and specific nested reverse transcription-PCR method using VHL gene mutation to detect circulating cancer cells. We screened 29 sporadic clear cell RCCs from patients for mutations of the VHL gene by direct sequencing. We next examined blood samples from patients with the VHL gene mutation using mutation-specific nested reverse transcription-PCR. Somatic mutations were detected in 20 of 29 (69.0%) sporadic clear cell RCCs. The VHL gene mutations were detected in peripheral and/or renal venous blood from 15 of 20 (75%) patients. The mutations were detected in the peripheral blood in 2 of 17 (11.8%) patients before surgery, 6 of 16 (37.5%) patients within 24 h after surgery, 3 of 16 (18.8%) patients on day 7 after surgery, and 2 of 11 (18.2%) patients on day 30 after surgery. In seven of nine (77.8%) patients, mutations were detected in renal venous blood during surgery. These findings indicate the presence of circulating cancer cells with VHL gene mutation. Although much larger studies are needed to determine the clinical significance, our study shows that this technique is feasible for detecting circulating RCC cells.

Overexpression of c-met proto-oncogene associated with chromophilic renal cell carcinoma with papillary growth.

Various genetic changes are involved in human renal cell carcinomas (RCCs). However, the molecular events related to other cytomorphological subtypes of RCC are not well known, apart from the relationship between the von Hippel-Lindau tumour suppressor gene and clear cell subtype RCC. We examined the overexpression of several growth factor receptors immunohistochemically and analyzed their relationship to the cytomorphological characters in 120 cases of RCCs. These receptors included c-met proto-oncogene product (c-MET), epidermal growth factor receptor (EGFR) and transforming growth factor beta receptor II (TGFbetaR). The overexpression of c-MET was detected in all cases (20/20) of the tubulo-papillary growth type and 78.3% (18/23) of chromophilic cell subtype, resulting in a very significant associations between c-MET overexpression and tubulo-papillary growth RCCs (P<0.0001), c-MET and chromophilic subtype RCCs (P<0.0001), and c-MET and EGFR (P<0.0001). EGFR overexpression was significantly associated with the compact growth RCCs (49/89, P<0.0001), clear cell subtype RCCs (P<0.005) and the overexpression of TGFbetaR (P<0.0001). These results strongly suggest a close correlation between the overexpression of c-MET and development of the chromophilic subtype of RCC with papillary growth pattern. EGFR expression is closely related to the pathogenesis of the clear cell subtype of RCC with compact growth pattern. The overexpression of c-MET, EGFR, and TGFbetaR may have roles that are individually significant in the morphogenesis of RCC.

Dermatofibrosarcoma protuberans harboring t(9;22)(q32;q12.2).

More than 20 cases of dermatofibrosarcoma protuberans (DFSP) exhibiting chromosomal abnormalities have been reported. Approximately three fourths of these tumors have harbored supernumerary ring chromosomes, which have been suggested to be specific for this tumor. However, a small number of DFSPs with translocations such as t(2;17), t(X;7), and t(17;22) have recently been reported. We report a DFSP arising in a 23-year-old woman which unexpectedly exhibited the balanced translocation, t(9;22)(q32;q12.2) as the only anomaly with G-band technique. Dual-color fluorescence in situ hybridization (FISH) confirmed these cytogenetic findings. Similar to that previously reported for DFSPs with translocations, the present tumor also lacked ring chromosomes.

Overexpression of c-Met/hepatocyte growth factor receptors in human prostatic adenocarcinoma.

Hepatocyte growth factor (HGF) and c-met proto-oncogene product (c-Met) have varied biological functions in different tissues and have been implicated in mitogenic, motogenic and morphogenic responses in both organ regeneration and carcinogenesis. Some studies have suggested that the overexpression of c-Met and epidermal growth factor receptor (EGFR) are associated with growth advantage, while transforming growth factor-beta receptor II (TGF beta R II) is associated with growth disadvantage of human prostatic adenocarcinoma. However, it is unclear if the expression of c-Met correlates with the expression of EGFR and TGF beta R II, and with the proliferative status of human prostatic adenocarcinoma. Using immunohistochemical staining with anti-c-Met (C-12), anti-EGFR (NCL-EGFR) and anti-TGF beta R II (L-21) antibodies, we determined the frequency of expression of c-MET, EGFR, and TGF beta R II respectively in a series of 134 radical prostatectomy specimens. We evaluated the relationship between the expression of these receptors and clinicopathological characteristics. Overall, c-Met immunostaining was detected in 54 of 134 (40.3%) cases, EGFR in 45 (33.6%) and TGF beta R II in 64 (48.4%). The overexpression of c-Met was significantly more common in poorly differentiated (P < 0.0001) and in the diffusely infiltrated specimens (P < 0.0005). In contrast, TGF beta R II was significantly overexpressed in the well differentiated specimens (P < 0.0001) and associated negatively with c-Met (P < 0.0001). Overall, these data suggest that c-Met/HGF receptor and TGF beta R II overexpression may be involved in the differentiation of human prostatic adenocarcinoma, c-Met with de-differentiation and TGF beta R II with differentiation.

[Enucleation of renal tumor using microwave tissue coagulator: a case report].

A 69-year-old man was referred to our department for a cystic tumor, 4.0 cm in diameter, in the lower portion of the right kidney, which was detected by computed tomography. The patient had been admitted to the department of surgery in our hospital for treatment of ileus caused by transverse colon cancer. With a diagnosis of cystic renal cell carcinoma. T2N0M0, in situ non-ischemic tumor enucleation was performed using a microwave tissue coagulator (Microtaze, Heiwa Electronics Industry Inc., Tokyo). The enucleation was accompanied by a defect of the renal pelvis, but it was easily repaired. The operation time was 120 minutes and blood loss was 110 cc. The histological diagnosis was renal cell carcinoma, pT2N0M0V1, expansive, alveolar type, clear cell subtype, G1 > G2. Diagnostic imaging done postoperatively showed no sign of damage to renal function. At the present time, the patient has been disease-free with interferon-alpha for 12 months and is being followed on an outpatient basis. In this report, the advantages of nephron-sparing surgery, especially in situ non-ischemic tumor enucleation using a microwave tissue coagulator for renal tumor are discussed. In particular, the technique of performing tumor enucleation with repair of the defect of renal pelvis used in this case may extend the indication of nephron-sparing surgery.

Origin of Differences in Heats of Immersion of Silicas in Water.

The origin of differences in immersional heats of various silicas in water has been investigated by measurements of water vapor adsorption isotherms, strength of electrostatic fields of surfaces, and concentration and infrared spectra of hydroxyl groups. The contribution of interaction energy between dipole moments of water molecules and the electrostatic fields of silica surfaces to immersional heat was 30-40% of the total heat of immersion. The differences in immersional heats of silicas in water significantly depend on the interaction of hydrogen bonds between water molecules and surface hydroxyl groups rather than on the interaction between dipole moments of water molecules and surface polarity. The hydrogen-bonded hydroxyl groups strongly interact with water molecules rather than the free hydroxyl groups and cause an increase in heat of immersion. Copyright 1998 Academic Press.

TPD, FTIR, and Molecular Adsorption Studies of Calcium Hydroxyapatite Surface Modified with Hexanoic and Decanoic Acids.

The surface of synthetic calcium hydroxyapatite Ca10(PO4)6 (OH)2 (CaHAP) particles was modified with hexanoic CH3(CH2)4 COOH (HA) and decanoic CH3(CH2)8COOH (DA) acids in hexane and the resulting materials were characterized by various means. XRD pattern, specific surface area, and morphology of the CaHAP particles were essentially not altered by the modification. FTIR and TPD results indicated that HA and DA molecules are hydrogen-bonded to the surface P-OH groups of CaHAP in three adsorption types. The surface of modified particles became hydrophobic. When the modified samples were outgassed at 500 degrees C, HA and DA were removed and the surface P-OH groups were revived. The modified material adsorbed much less H2O and CO2 than the unmodified one. Copyright 1999 Academic Press.

Surface Structure and Properties of Calcium Hydroxyapatite Modified by Hexamethyldisilazane.

The surface of synthetic calcium hydroxyapatite Ca10(PO4)6(OH)2 (CaHAP) particles was treated by repeated modification with hexamethyldisilazane [(CH3)3Si]2NH (HMDS) in hexane and thermal treatment and the surface of the modified CaHAP was characterized by various means. No remarkable change in XRD patterns or in particle shape by the modification was observed. The width of the CaHAP particles gradually increased with repeating the modification. FTIR results indicated that HMDS reacted with surface P-OH groups of CaHAP to yield surface Si-(CH3)3 groups. The surface of the modified CaHAP was hydrophobic. The surface Si-(CH3)3 groups turned to three kinds of surface Si-OH groups by treating the modified materials at 500 degreesC in air. These formed surface Si-OH groups and the remaining surface P-OH groups reacted with HMDS by repeating the modification, resulted in the increase of the surface Si atoms. The modified material having surface Si-(CH3)3 or Si-OH groups adsorbed much less CO2 than the unmodified one. Copyright 1998 Academic Press.