Hypertrophy of rabbit proximal tubule cells is associated with overexpression of TGF beta

The expression of transforming growth factor beta (TGF beta) in proximal tubule cells from rabbit kidney cortex after uninephrectomy (UNX) was investigated. Cell protein and [3H]leucine incorporation in these cells were significantly increased, while cell number was decreased, at two weeks following UNX. At this time period after UNX, we found that proximal tubule cells showed a dramatic increase of cytoplasmic immunostaining with a pan-specific anti-TGF beta antibody. This was accompanied by a 3-fold increase in TGF beta 1 mRNA expression in these cells. Furthermore, proximal tubule cells from two-week uninephrectomized rabbits secrete about 2-fold higher TGF beta bioactivity to the cell conditioned medium compared to cells from sham-operated animals. Addition of anti-TGF beta 1, beta 2, beta 3 neutralizing antibody increased the growth of the former cells, and it abolished cell hypertrophy. These results indicate that hypertrophy of proximal tubule cells during compensatory renal growth is associated with overexpression of TGF beta.     

A phase II study of DaunoXome in advanced urothelial transitional cell carcinoma

Liposomal encapsulation of anthracyclines is claimed to reduce toxicity and to improve pharmacokinetics. Therefore, 15 patients with locally advanced or metastatic transitional cell cancer (TCC) of the urinary tract were entered into a phase II study assessing the response rate (WHO criteria) and toxicity of DaunoXome 100 mg/m2 given as a 1 h infusion every third week. During treatment, 6 patients remained stable and 8 had progressive disease. 1 patient died of pulmonary embolism after the first cycle and was not evaluable for response. No patient developed grade 4 myelotoxicity. Grade 3 leucopenia was seen in 5 patients and grade 1 thrombocytopenia in 1 patient, with no treatment-related changes of biochemical liver and kidney function tests. 4 patients complained of angina pectoris-like chest pain during the initial phase of the first or second infusion. The event was associated with a decrease in systolic blood pressure by 20-30 mm in 1 patient leading to permanent treatment discontinuation. In the other 3 and all subsequent patients, intramuscular application of 100 mg hydrocortisone 1 h prior to DaunoXome infusion prevented similar hypotensive reactions. In this study, intravenous (i.v.) DaunoXome 100 mg/m2 every third week showed no anticancer activity in advanced TCC.     

Epidermal growth factor receptor expression is associated with rapid tumor cell proliferation in renal cell carcinoma

Epidermal growth factor receptor (EGF-r) expression and tumor cell proliferation rate have been proposed as potential prognostic parameters in renal cell carcinoma (RCC). In this study, immunohistochemical stains using antibodies to EGF-r and the cell proliferation marker Ki-67 (MIB-1) were used to study the relationship between EGF-r expression, tumor cell proliferation, and prognosis in 50 non-papillary RCC extending beyond the renal capsule (pT3). A high Ki-67 labeling index (LI) was associated with poor patient prognosis (P < .05). Thirty-eight cases (76%) expressed strong cell membrane immunoreactivity for EGF-r. There was a tendency toward a shortened survival for EGF-r-positive tumors (P = .08). Tumor growth fraction (Ki-67 LI) was significantly higher in EGF-r-positive tumors than in EGF-r-negative tumors (P < .05), suggesting that rapid tumor proliferation might be responsible for the poor prognosis associated with EGF-r-positive RCC.     

Reduced E-cadherin expression correlates with increased invasiveness in colorectal carcinoma cell lines

A reduction in cell adhesiveness and cell invasion are essential steps in tumour progression to metastasis. In the present study two out of seven colorectal carcinoma cell lines exhibited reduced expression of the cell-cell adhesion molecule E-cadherin as assessed by immunofluorescence. The same two cell lines were invasive in the collagen gel and membrane invasion culture system invasion assays. Addition of anti-E-cadherin antibody to a non-invasive carcinoma cell line caused the cells to assume a dissociated morphology on plastic and to become invasive in collagen gels. This demonstrates a causal role for E-cadherin in the maintenance of intercellular adhesion and the suppression of tumour cell invasion and possibly metastasis in colorectal tumour cells.     

Reduced expression of cyclin-dependent kinase inhibitor p27Kip1 is associated with advanced stage and invasiveness of gastric carcinomas

Reduced expression of a cyclin-dependent kinase inhibitor p27Kip1 has recently been shown to predict poor survival of patients with breast and colorectal cancers. We studied the expression of p27Kip1 in gastric carcinomas by northern blotting, western blotting and immunohistochemistry to determine whether lack of p27 has implications for aggressiveness of gastric cancer. Reduced expression of p27 was detected in 40% of the gastric carcinomas at the mRNA level, while it was detected in 57% at the protein level. No gross alterations of the p27 gene were observed in any of the cases examined by Southern blot analysis. Immunohistochemical studies revealed that the expression of p27 was well preserved in most of the gastric adenomas, whereas it was so in only 26% of the gastric carcinomas. Fifty-six percent of the carcinomas showed almost no p27-positive cells. Decrease of p27-positive cells significantly correlated with advanced stage, depth of tumor invasion and lymph node metastasis. The expression of p27 showed an inverse correlation with the expression of cyclin E. These findings suggest that reduction of p27Kip1 protein may reflect the progression of gastric carcinomas and may be an indicator of high-grade malignancy.     

Urinary bladder transitional cell carcinoma with trophoblastic differentiation

Transitional cell carcinoma (TCC) with trophoblastic differentiation (TD) is a newly recognized variant of urothelial cancer which produces placental proteins, predominantly beta-human chorionic gonadotropin (HCG). It has a poor prognosis. About 210 cases were described, mostly from North America, Europe and Japan. This is the first report of TCC TD in a resident of Israel's upper Galilee. A 69-year-old man whose urinary papillary bladder tumor was established cystoscopically, refused treatment and stopped follow-up. 3.5 years after his last visit, he returned and cytologic examination revealed malignant urothelial cells, while intravenous pyelography disclosed a urinary bladder defect. Cystoscopy showed numerous papillary masses dispersed over the bladder mucosa, which were resected transurethrally. Histopathologic examination revealed TCC grade III, stage A. Tumor cells were immunopositive for beta-HCG and human placental lactogen. 4 transurethral resections of large masses were performed within 2 months. Pulmonary metastases developed and the patient died 4 years after the detection of the urinary bladder tumor.     

Opposite effect of stable transfection of bioactive transforming growth factor-beta 1 (TGF beta 1) versus exogenous TGF beta 1 treatment on expression of 92-kDa type IV collagenase in mouse skin squamous cell carcinoma CH72 cells

We have previously shown that transforming growth factor-beta 1 (TGF beta 1) mRNA is consistently overexpressed in squamous cell carcinomas relative to normal mouse skin. Here we show that 92-kDa type IV collagenase (matrix metalloproteinase) (MMP-9) mRNA was likewise progressively overexpressed during mouse skin carcinogenesis. To determine if overexpression of MMP-9 and TGF beta 1 are linked, we stably transfected a bioactive TGF beta 1 into a mouse skin squamous cell carcinoma cell line (CH72), which resulted in about twofold to three-fold higher levels of secreted active TGF beta 1. Active TGF beta 1-transfected cells grew only slightly, but not significantly, more slowly in vitro and in vivo than vector-only transfectants. Two clones overexpressing active TGF beta 1 secreted much reduced levels of MMP-9 activity, as determined by zymogram analyses. However, treatment of these clones with 40 pM exogenous TGF beta 1 for 48 h enhanced secretion of MMP-9 activity. Constitutive mRNA expression of MMP-9 was reduced twofold to 70-fold in five untreated active TGF beta 1-transfected clones relative to the other transfectants. In contrast, treatment with 40 pM exogenous TGF beta 1 induced MMP-9 mRNA expression in a time-dependent fashion, from twofold to fourfold after 4 h to a maximum of 12- to 19-fold after 24-48 h. Induction of MMP-9 mRNA was dose dependent at TGF beta 1 concentrations of 4-400 pM. Thus, stable transfection of bioactive TGF beta 1 downregulated whereas exogenous TGF beta 1 treatment upregulated MMP-9 activity and expression. Treatment of transfectants with a neutralizing TGF beta 1 antibody slightly downregulated constitutive MMP-9 mRNA (20-30%) but completely blocked induction by exogenous TGF beta 1. Thus, the effect of TGF beta 1 transfection was not due to secreted TGF beta 1 but may have been a secondary effect.     

Primary transitional cell carcinoma in vesical diverticula

OBJECTIVES: To evaluate the treatment and prognosis of primary tumors in bladder diverticula. METHODS: The cases of 611 patients treated for bladder tumors at a single medical center were retrospectively reviewed. RESULTS: Eight patients had primary intradiverticular transitional cell carcinoma. Five patients had Stage Ta tumor, and 3 had Stage T1 tumor. Most patients were treated by local resection and adjuvant intravesical chemotherapy. All patients with initial Ta disease are disease free at the time of this writing. One patient with T1 disease died, 1 patient's disease recurred several times, and 1 patient showed positive cytology without apparent disease. CONCLUSIONS: Superficial intradiverticular tumors may be treated conservatively. Routine cystoscopy for patients with a bladder diverticulum is warranted for early diagnosis of possible intradiverticular tumor.     

Doxorubicin hydrochloride, cyclophosphamide, and 5-fluorouracil combination in advanced prostate and transitional cell carcinoma

Hubbard cockerels, 1 day of age (n = 91), were divided into 3 handling groups: (1) chicks from the 5-Day group were removed from the brooder and isolated at 22 degrees C for 3 min on each of the first 5 days; (2) chicks from the 15-Day group received this treatment for 15 days; and (3) Control chicks were not handled. Within each group half were fed broiler starter for 5 weeks, and half were fed layer starter which produces slower growth. On Days 28, 35, 42, and 49 the average weight of 5-Day birds was significantly higher than that of Controls; 15-Day birds weighed slightly less than birds in the 5-Day group. The weight advantage of the 5-Day birds occurred with both starter rations, at all weighing periods, and did not appear to reflect increased food consumption. Similarities between these findings and results obtained with rodents were noted.     

Sialosyl Tn antigen expression is associated with the prognosis of patients with advanced gastric cancer

BACKGROUND: Several studies have revealed a correlation between sialosyl Tn antigen (STN) and certain clinicopathologic features of various cancers, and that STN is an independent prognostic factor. However, the clinical significance of the expression of STN in gastric cancer has not been reported. Thus, the purpose of this study was to evaluate immunohistochemically the clinical significance of expression of STN in gastric cancer. METHODS: The expression of STN in surgically resected specimens of human gastric cancer was evaluated immunohistochemically using a monoclonal antibody (TKH-2), in 60 patients whose serum STN levels were measured and in 54 patients with advanced cancer who had been followed for more than 5 years after gastrectomy. The correlations between the level of STN expression and clinicopathologic factors were analyzed. The staining intensity was graded as follows: (-), less than 5% of the cancer cells expressed STN; (+), 5-50%; (++), more than 50%. RESULTS: Sialosyl TN antigen staining was detected mainly on the cell membrane, in the cytoplasm, and in the luminal contents, and 57.2% of the 60 specimens expressed STN, whereas the corresponding value for positive serum levels was 15%. A higher percentage of advanced tumors expressed STN than did the early cases, but the difference was not statistically significant. All cases with strong staining, the (++) cases, were advanced cases either with lymph node metastases or with cancer invading in or beyond the muscle layer proper. The expression of STN appeared to be related to the clinical stage, the extent of cancer invasion, and the presence of lymph node metastases. Sialosyl TN antigen was detected in the serum in less than 6% of the patients whose tumors were (-) or (+) for STN expression, and in 86.7% of the patients whose tumors expressed high levels of STN (++). The estimated 5-year survival in advanced cases (Stage III) was significantly better in those with negative STN expression than in those with positive STN expression (P < 0.01). CONCLUSIONS: These results suggest that STN may be a useful marker associated with the prognosis of patients with advanced gastric cancer.     

Reduced expression of the ICE-related protease CPP32 is associated with radiation-induced cisplatin resistance in HeLa cells

Low-dose fractionated gamma-irradiation (three cycles of 5 x 2 Gy) induced cisplatin resistance in HeLa cells. The drug resistance was modest (Rf of about 2) and stable, similar to that found previously in murine cells after irradiation. In the drug-resistant HeLa-C3 cells, flow cytometric analysis revealed a decreased number of apoptotic cells compared with the parental cells. Drug resistance was associated with considerably enhanced expression of the p53 suppressor protein in HeLa-C3 cells after cisplatin exposure but seemed not to be regulated by the bcl-2-dependent pathway. Cisplatin resistance correlated with reduced expression of ICE-related proteases (interleukin-1beta-converting enzyme). Basal levels of the 45-kDa precursor ICE protein were reduced in HeLa-C3 cells, while those of the mature 60-kDa heterotetramer were similar. The CPP32 protease, a member of the ICE family with structural homology but different substrate specificity, was expressed at a lowered level. After drug exposure, there was a slight increase of CPP32 in HeLa-C3 cells, equivalent to about 45% of the level attained in the parental cells. This is in contrast to the CPP32 levels measured after irradiation, which were similar in sensitive and in resistant cells. As the radiosensitivity is unchanged in both cell lines, these results suggest that cisplatin resistance in HeLa-C3 cells is associated with alterations of a CPP32-linked apoptotic pathway, which is affected by the damage caused by cisplatin but not by irradiation. Whether these changes are dependent on the observed p53 modifications is now being studied in resistant clones.     

Percutaneous management of upper-tract transitional cell carcinoma

Standard management of upper tract transitional cell carcinoma is a nephroureterectomy; however, there is a distinct subgroup of patients who would be rendered dialysis dependent, or at high risk for functionally significant renal insufficiency, and for whom a nephron-sparing approach is warranted. Historically, the options for such "conservative" management have been limited to variations of open pyelotomy or partial nephrectomy. More recently, endourologic techniques have been applied to the treatment of these difficult patients. Though a ureteroscopic approach can occasionally be therapeutic, it may be precluded by the size or location of the tumor, or by a urinary diversion that makes ureteroscopic access difficult or impossible. Many of these patients can, however, be managed with a well planned percutaneous approach that provides excellent access to virtually the entire upper tract. The subsequent addition of topical BCG as an adjunct to percutaneous upper tract tumor resection may then be beneficial in decreasing the incidence of local recurrence.     

Cytogenetic analysis in transitional cell carcinoma of the bladder

The potential use of numerical chromosomal abnormalities as predictive factors for the clinical behaviour of transitional cell carcinoma (TCC) was investigated. The effects on survival and progression-free survival were measured in 91 patients with TCC treated by transurethral resection. The survival rate of patients having tumours with a diploid chromosomal modal number was significantly better than that of patients having tumours with a hyperdiploid chromosomal modal number. The survival rate of patients having TCC with diploid cells only was also significantly better than that of patients having TCC with both diploid and hyperdiploid cells. Progression-free survival was significantly higher in patients having TCC with a diploid modal number of chromosomes than in patients with a hyperdiploid modal number. Simultaneous evaluation of the modal chromosome number or chromosomal range, histological grade, category and mitotic index of the tumour, and the patient's age and sex as prognostic factors in multivariate analyses showed that the category of bladder carcinomas was the most important factor in predicting the survival rate. In patients with superficial tumours (category Ta and T1) the modal chromosome number was the most important factor in predicting survival. For progression-free survival, the modal chromosome number appeared to be the most important factor. It was concluded that the modal chromosome number in TCC was useful in predicting survival in patients with superficial tumours and in predicting progression-free survival in patients with tumours of all categories.     

CD44 variant expression in inflammatory colonic mucosa is not disease specific but associated with increased crypt cell proliferation

The implementation of effective methods of testing the precision of computations, is essential for the development of medical systems, such as the stereotactic planning software for neurosurgery. During the development of our planning system we designed, in addition to the traditional phantom test, some tests based on digitally constructed images. These tests give quantitative and qualitative measurement of the precision and stability of the calculations, and are easy to implement and isolate the program error from the mechanical and image acquisition errors. The verification of our software shows good precision in coordinates calculation and a significant impact of pixel size and interscan spacing on the precision.     

In oesophageal squamous cell carcinoma vascular endothelial growth factor is associated with p53 mutation, advanced stage and poor prognosis

Vascular endothelial growth factor (VEGF) affects malignant tumours by promoting angiogenesis. The tumour-suppressor gene p53 has been thought to regulate VEGF. We investigated the effect of VEGF on oesophageal carcinoma and the connection between VEGF and p53. One hundred and nine resected oesophageal squamous cell carcinomas were examined. VEGF expression was analysed by immunohistochemical staining. Sixty-five tumours (59.6%, 65 out of 109) were classified as VEGF positive. A significant correlation was found between the VEGF expression and both the depth of invasion (P = 0.0001) and lymph node metastasis (P < 0.0001). With regard to p53, we compared the expression of VEGF with the mutation of p53, examined using polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and direct sequencing in tumour samples obtained from 36 patients who we have reported previously. The VEGF expression was significantly correlated to p53 mutation (P = 0.0291). To evaluate the angiogenesis, microvascular density (MVD) was counted, and endothelial cells were stained immunohistochemically using anti-CD34 monoclonal antibody against 29 cases with invasion limited to the submucosal layer. The average MVD had a tendency to correlate to VEGF expression (P = 0.1626). The prognoses of patients with VEGF-positive primary tumours were significantly worse than for those with VEGF-negative primary tumours (P = 0.0077). We have assumed that VEGF contributes to aggressive characteristics in oesophageal carcinomas and that VEGF expression might be affected by p53 status.