Nonpapillary Renal Cell Carcinomas Research Articles

Clear cell and papillary renal cell carcinomas in hereditary papillary renal cell carcinoma (HPRCC) syndrome: a case report

BackgroundHereditary papillary renal cell carcinoma (HPRCC) is a rare autosomal dominant disease characterized by the development of multiple and bilateral papillary type I renal cell carcinomas (RCC) and papillary adenomas caused by activating mutations in the MET proto-oncogene. Classically, distinctive histological features of RCC are described according to the familial renal cell carcinoma syndrome. To date, no clear cell RCC has been reported in HPRCC syndrome.Case presentationWe describe the case of a 51-year-old man with a germline MET mutation detected on peripheral blood testing, and no germline VHL mutation, who developed numerous papillary tumors but also unexpectedly clear cell renal cell carcinomas. During the follow-up, an adrenal metastasis was observed 7 years after the initial diagnosis corresponding to a clear cell RCC metastasis. By immunohistochemistry, clear cell tumors showed focal cytokeratin 7, moderate racemase, and diffuse and membranous CAIX expression, while papillary tumors expressed strong diffuse cytokeratin 7 and racemase without CAIX positivity. Using FISH, VHL deletion was observed in one of the clear cell tumors, and the metastatic clear cell tumor presented a trisomy of chromosomes 7 and 17. These last genomic alterations are usually detected in papillary RCC, highlighting the potential link between both histological subtypes of tumors and the HPRCC syndrome.ConclusionsThe pathologist must be aware that the presence of a non-papillary RCC associated with numerous papillary tumors should not exclude the diagnostic suspicion of HPRCC and thus to perform a thorough genomic study.

The correlation between gain of chromosome 8q and survival in patients with clear and papillary renal cell carcinoma.

The proto-oncogene c-MYC, located on chromosome 8q, can be upregulated through gain of 8q, causing alteration in biology of renal cell carcinoma (RCC). The aim of this study was to evaluate the prevalence of c-MYC through chromosome 8q gain and to correlate findings with cancer-specific mortality (CSM), and overall survival (OS). Cytogenetic analysis by conventional or Chromosomal Genomic Microarray Analysis (CMA) was performed on 414 renal tumors. Nonclear and nonpapillary RCC were excluded. Impact of gain in chromosome 8q status on CSM, OS, and its correlation with clinicopathological variables were evaluated. CSM and OS were assessed using log-rank test and the Cox proportional hazards model. A total of 297 RCC tumors with cytogenetic analysis were included. Gain of 8q was detected in 18 (6.1%) tumors (9 clear cell and 9 papillary RCC), using conventional method (n = 11) or CMA (n = 7). Gain of 8q was associated with higher T stage (p < 0.001), grade (p < 0.001), nodal involvement (p = 0.005), and distant metastasis (p < 0.001). No association between gain of 8q and age (p = 0.23), sex (p = 0.46), and Charlson comorbidity index (CCI, p = 0.59) were seen. Gain of 8q was associated with an 8.38-fold [95% confidence interval (CI), 3.83-18.34, p < 0.001] and 3.31-fold (95% CI, 1.56-7.04, p = 0.001) increase in CSM and decrease in OS, respectively, at a median follow up of 56 months. Chromosome 8q harbors the proto-oncogene c-MYC, which can be upregulated by gain of 8q. Our findings suggest that gain of 8q, can predict aggressive tumor phenotype and inferior survival in RCC.

MP29-04 GAIN OF CHROMOSOME 8Q IS ASSOCIATED WITH INFERIOR SURVIVAL IN PATIENTS WITH RENAL CELL CARCINOMA

You have accessJournal of UrologyKidney Cancer: Basic Research II1 Apr 2014MP29-04 GAIN OF CHROMOSOME 8Q IS ASSOCIATED WITH INFERIOR SURVIVAL IN PATIENTS WITH RENAL CELL CARCINOMA Reza Mehrazin, Robert G. Uzzo, Essel Dulaimi, Karthik Devarjan, Jeffrey J. Tomaszewski, Joseph Testa, Jianming Pei, Philip Abbosh, Timothy Ito, Alexander Kutikov, and Tahseen Al-Saleem Reza MehrazinReza Mehrazin More articles by this author , Robert G. UzzoRobert G. Uzzo More articles by this author , Essel DulaimiEssel Dulaimi More articles by this author , Karthik DevarjanKarthik Devarjan More articles by this author , Jeffrey J. TomaszewskiJeffrey J. Tomaszewski More articles by this author , Joseph TestaJoseph Testa More articles by this author , Jianming PeiJianming Pei More articles by this author , Philip AbboshPhilip Abbosh More articles by this author , Timothy ItoTimothy Ito More articles by this author , Alexander KutikovAlexander Kutikov More articles by this author , and Tahseen Al-SaleemTahseen Al-Saleem More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.746AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The proto-oncogene c-MYC, located on chromosome 8q, can be up-regulated through gain of 8q causing alteration in biology of renal cell carcinoma (RCC). The aim of this study was to evaluate the prevalence of chromosome 8q gain in our renal cancer tumor registry and to correlate findings with tumor histology, grade, cancer specific survival (CSS), and overall survival (OS) in clear and papillary METHODS Cytogenetic analysis by conventional or Chromosomal Microarray Analysis (CMA) was performed on tumors of 414 patients who underwent radical or partial nephrectomy for suspected RCC. Non-clear RCC, non-papillary RCC, and those which did not show alteration or gain in chromosome 8 on cytogenetic analysis were excluded from the study. Impact of gain in chromosome 8q status on CSS, OS, and its correlation with clinicopathological variables, obtained from our prospectively maintained kidney cancer database, were evaluated and compared to RCC’s that did not have gain in chromosome 8q. CSS and OS were assessed using log-rank test and the Cox proportional hazards model. RESULTS After the exclusions total 279 clear cell and papillary RCC tumors with cytogenetic analysis were included in study. Gain of 8q was detected in 18 (6.5%) tumors (9 clear cell, 3 sarcomatoid variant, and 6 papillary RCC), using either conventional method (11) or CMA (7). Gain of 8q was associated with higher grade (12% vs 2.0%, p = 0.001), higher risk of regional lymph node involvement (30 % vs. 6%, p=0.008), and distant metastasis (22 % vs. 4.0%, p<0.001). Among tumors with 8q gain, 56% were stage III or higher. No association between gain of 8q and age (p=0.23), sex (p=0.42), and CCI (p=0.59) was seen. Gain of chromosome 8q was associated with a 6.56 (95% CI, 3.06-14.05, p<0.0001) and 2.48 (95% CI, 1.33-5.92, p=0.007) fold decrease in CSS and OS, respectively, at a median follow-up of 56 months. CONCLUSIONS Chromosome 8q harbors the proto-oncogene c-MYC, which can be up regulated by gain of 8q. Our findings suggest that gain of 8q, shown by cytogenetics, can predict aggressive tumor phenotype and inferior survival in patients with RCC. These findings may help clinicians to identify high risk patients and enroll them into neoadjuvant and adjuvant clinical trials. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e305-e306 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Reza Mehrazin More articles by this author Robert G. Uzzo More articles by this author Essel Dulaimi More articles by this author Karthik Devarjan More articles by this author Jeffrey J. Tomaszewski More articles by this author Joseph Testa More articles by this author Jianming Pei More articles by this author Philip Abbosh More articles by this author Timothy Ito More articles by this author Alexander Kutikov More articles by this author Tahseen Al-Saleem More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Utility and limitations of 3-Tesla diffusion-weighted magnetic resonance imaging for differentiation of renal tumors

ObjectiveTo investigate utility and limitations of 3-Tesla diffusion-weighted (DW) magnetic resonance imaging (MRI) for differentiation of benign versus malignant renal lesions and renal cell carcinoma (RCC) subtypes. Materials and methodsSixty patients with 71 renal lesions underwent 3 Tesla DW-MRI of the kidney before diagnostic tissue confirmation. The images were retrospectively evaluated blinded to histology. Single-shot echo-planar imaging was used as the DW imaging technique. Apparent diffusion coefficient (ADC) values were measured and compared with histopathological characteristics. ResultsThere were 54 malignant and 17 benign lesions, 46 lesions being small renal masses ≤4cm. Papillary RCC lesions had lower ADC values (p=0.029) than other RCC subtypes (clear cell or chromophobe). Diagnostic accuracy of DW-MRI for differentiation of papillary from non-papillary RCC was 70.3% resulting in a sensitivity and specificity of 64.3% (95% CI, 35.1–87.2) and 77.1 (95% CI, 59.9–89.6%). Accuracy increased to 83.7% in small renal masses (≤4cm diameter) and sensitivity and specificity were 75.0% and 88.5%, respectively. The ADC values did not differ significantly between benign and malignant renal lesions (p=0.45). ConclusionsDW-MRI seems to distinguish between papillary and other subtypes of RCCs especially in small renal masses but could not differentiate between benign and malignant renal lesions. Therefore, the use of DW-MRI for preoperative differentiation of renal lesions is limited.

Differential Diagnosis of Renal Tumors With Papillary Architecture

Papillary renal cell carcinoma is the second most common malignant renal epithelial tumor and constitutes approximately 15% of renal cell tumors. However, papillary architecture is neither unique to papillary renal cell carcinoma, nor do all papillary renal cell carcinomas show exclusive papillary histology. Many of the nonpapillary renal cell carcinomas with papillary architecture have been recognized only recently. Distinction of these from papillary renal cell carcinoma is essential, as biologic behavior and potential therapeutic options are distinct in many such tumors. Close attention to the cytologic and growth pattern characteristics will allow us to arrive at the proper diagnosis in most cases, although sometimes immunohistochemistry and rarely genetic evaluation may be needed.

Association of the BCRP C421A polymorphism with nonpapillary renal cell carcinoma

Breast cancer resistance protein (BCRP), the second member of the ATP-binding cassette membrane transporter family, has a single nucleotide polymorphism, C421A (resulting in Q141K), that is of functional importance. Our aim was to explore the relationship between this polymorphism of the BCRP gene and the risk of renal cell carcinoma (RCC) development. For a case-control study, DNA samples from 200 nonpapillary RCC patients and 200 healthy control subjects were analyzed using the TaqMan technique. The genotypic frequencies of the BCRP C421A polymorphism were compared between RCC patients and control subjects. The frequency of the C/C genotype was significantly higher in RCC patients than in control subjects (age- and gender-adjusted OR = 1.96, 95% CI 1.32-2.93). No associations were observed between the BCRP C421A polymorphism and clinicopathologic or epidemiologic factors, including age, gender, tumor grade, stage, cigarette smoking, family history of cancer and body mass index. Carriers with the C/C genotype of the BCRP C421A polymorphism are at risk of developing nonpapillary RCC. These data suggest that BCRP is a candidate RCC susceptibility gene.

Pediatric Renal Carcinoma Associated with Xp11.2 Translocations/TFE3 Gene Fusions and Clinicopathologic Associations

Renal cell carcinomas (RCCs) are rare in children and studies of their subtypes and clinicopathologic associations are limited to small series. We identified 8 patients with RCC treated at our institution between 1981 and 2003, reviewed their clinicopathologic features, cytogenetics findings, and evaluated the status of TFE3 expression by immunohistochemistry and numerical chromosomal alterations by interphase fluorescent in situ hybridization on paraffin-embedded tissue. These 8 patients (5 female and 3 male) had diploidy, and 5 had morphologic features compatible with the recently described RCC associated with Xp11.2 translocations/TFE3 gene fusions and demonstrated nuclear labeling for TFE3 protein by immunohistochemistry. The translocation was confirmed in 2 of these 5 patients by conventional cytogenetics. One case was a high-grade nonpapillary RCC and the other was compatible with type 2 papillary RCC. Four patients showed at least 1 chromosomal gain including trisomy 7 and/or trisomy 17. None of the tumors from male patients showed evidence of loss of the Y chromosome, but 2 patients showed numerical abnormalities of X chromosome +add(X). Two patients had sickle cell disease, and 1 of these also had stage IV-S neuroblastoma. This study suggests that many cases of RCC in children reported under the terms "papillary" and "clear cell" likely represent Xp11.2 translocation/TFE3 gene fusion-associated RCC. It also emphasizes the unusual associations of RCC with neuroblastoma and sickle cell hemoglobinopathy, which need further study.

Positive correlation between allelic loss at chromosome 14q24-31 and poor prognosis of patients with renal cell carcinoma

Objectives To report our development of a new application of the inter- Alu long polymerase chain reaction (PCR) for genomic scanning to screen for tumor-specific alterations in tumor DNA. Using this method, we detected a rearranged chromosomal region in renal cell carcinomas (RCCs). We then examined tumor-specific allelic loss in this region using microsatellite markers and determined whether a relationship was present between this allelic loss and the clinicopathologic features of the patients. Methods The inter- Alu long PCR genomic scan method was performed using RCC DNA samples and primers specific for a minor subset of the human repeat sequence Alu. We analyzed DNA samples from 42 pairs of matched normal and nonpapillary RCC tissues with seven microsatellite markers. Results The inter- Alu long PCR genomic scan method revealed an altered DNA region on chromosome 14q24-31, which is the location of several putative tumor suppressor genes. At least one of seven microsatellite markers on chromosome 14q24-31 showed loss of heterozygosity in 23 (54.8%) of 42 informative cases of RCC. The prevalent loss region was confined to a 2-Mb region around D14S67. We found a positive correlation between the presence of the loss of heterozygosity on 14q24-31 and tumor stage ( P <0.05). We also found that cases with allelic loss at 14q24-31 had a poor prognosis ( P = 0.045). Conclusions Our inter- Alu long PCR genomic scan method is a powerful method for the screening of DNA alterations, and our data suggest that the chromosome 14q24-31 region contains likely tumor suppressor genes associated with the progression of RCC.

Renal cell carcinoma: Allelic loss at chromosome 9 using the fluorescent multiplex–polymerase chain reaction technique

Loss of tumor-suppressor genes on both arms of chromosome 9 appears to be common in many types of cancer. Chromosome 9q is often partially deleted in bladder cancer, lung cancer, and basal cell carcinoma. However, little data are available on allelic loss on chromosome 9 in renal cell carcinoma (RCC). One hundred and nine nonpapillary RCCs were studied for loss of heterozygosity (LOH) at 13 loci on chromosome 9 by using the fluorescent multiplex–polymerase chain reaction method to compare DNA from tumor samples and peripheral blood lymphocytes. At the loci tested, LOH was found in from 2.3% (9q31, D9S938) to 17% (9q22, 1AJL) of informative cases, and 27 (24.8%) of the 109 RCCs had LOH at 1 or more loci of chromosome 9. LOH was more often detected at 9q22 within the PTCH gene (17%) when compared with LOH at the other 12 loci (P = 0.0172). Regarding the relationship with clinical parameters, however, there were no statistically significant associations between this LOH and tumor stage or grade. Among the 109 tumors, 6 (5.5%) showed replication errors. Our results suggest that LOH of the PTCH gene may be related to the development of nonpapillary RCC, although the clinical relevance has not been not clarified. HUM PATHOL 33:910-914. Copyright 2002, Elsevier Science (USA). All rights reserved.

Clinical significance of allelic loss of chromosome region 5q22.3∼q23.2 in nonpapillary renal cell carcinoma

To analyze the clinical significance of copy number gain and loss at chromosome region 5q21∼q23, 105 nonpapillary renal cell carcinomas (RCC) were examined by interphase cytogenetic analysis using the dual-color fluorescence in situ hybridization (FISH) technique. DNA probes for D5S23 (5p15.2), cCI5-243 (5q21.2∼q21.3), and cCI5-215 (5q22.3∼q23.2) were used, and the signals for cCI5-243 and cCI5-215 were compared with those for D5S23 as the numerical control. Aneusomy (three or more copies) of chromosome 5 was found in 22 tumors (21.0%). Aneusomy was significantly correlated with loss at 5q21∼q23, while disomy with gain at 5q21∼q23 ( P<0.05). Aneusomy was also significantly related to poor disease-specific survival ( P<0.01). Gain and loss at cCI5-243 were seen in 34 (32.4%) and 59 (56.2%) tumors, respectively, while gain and loss at cCI5-215 occurred in 55 (52.4%) and 45 (42.9%) cases, respectively. The frequency of gain at cCI5-215 was significantly correlated with a smaller tumor diameter (7 cm or less, P<0.05), while loss with a larger one (>7 cm, P<0.05). Both loss at cCI5-215 and aneusomy of chromosome 5 were significantly related to poor disease-specific survival ( P<0.05). In conclusion, alterations of chromosome 5 (including allelic loss of 5q22.3∼q23.2) could be a useful genetic marker for predicting the patient prognosis of RCC.

Introducing the Clusterin Gene Into Human Renal Cell Carcinoma Cells Enhances Their Metastatic Potential

We recently reported a protective role of clusterin expression against apoptosis induced by a wide variety of stimuli in several human cancer models. In the current study we tested the hypothesis that clusterin over expression confers a benefit for the metastasis of renal cell carcinoma through the inhibition of apoptosis induced by the various obstacles the cancer cells may confront after detachment from their primary origin. We introduced clusterin complementary DNA into human renal cell carcinoma ACHN cells, which do not express detectable level of clusterin expression, and generated the clusterin over expressing cell line ACHN/CL and the control vector only transfected cell line ACHN/C. In vitro anti-cell death activity under anchorage independent conditions among ACHN sublines was examined by limiting dilution assay and cell survival assay in suspension. To investigate the in vivo effects of clusterin over expression on metastatic potentials each cell line was injected into the tail vein or renal subcapsule of nonobese diabetic, severe combined immunodeficient mice and the metastatic features in all abdominal and thoracic organs were evaluated. ACHN/CL showed significantly enhanced growth in limiting dilution cultures compared with ACHN/C. The analysis of cell survival in the floating assay also revealed that ACHN/CL had a powerful survival advantage in suspension compared with ACHN/C. Furthermore, ACHN/CL formed more than 5-fold as many metastatic nodules in the lung after intravenous injection than ACHN/C. Similarly more marked lung metastasis was observed after implanting ACHN/CL cells into the renal subcapsule than after implanting ACHN/C cells. In contrast, there were no significant differences among ACHN sublines in the growth rates in vitro and in vivo, cell motility or invasive ability. These findings suggest that, if clusterin is over expressed, it prolongs cell survival under unfavorable conditions in the metastatic process, resulting in the enhanced metastatic potential of renal cell carcinoma.

OVER EXPRESSION OF CLUSTERIN IS AN INDEPENDENT PROGNOSTIC FACTOR FOR NONPAPILLARY RENAL CELL CARCINOMA

Recent studies have revealed the powerful anti-apoptotic activity of clusterin in several types of malignant tumors. However, the significance of clusterin expression in human renal cell carcinoma has not been well characterized. We determined whether the expression level of clusterin in nonpapillary renal cell carcinoma may be used as predictor of tumor progression and prognosis. Total RNA was extracted from 93 nonpapillary renal cell carcinomas obtained at surgery and expression levels of clusterin messenger (m)RNA in these specimens were measured by Northern blot analysis. Findings were analyzed with respect to several clinicopathological factors. There were 48 cases (52%) with strong clusterin mRNA expression and 45 (48%) in which it was weak or nonexistent. The expression level of clusterin mRNA did not significantly correlate with patient gender, age, tumor grade or size, or histological cell type, whereas pathological stage and the incidence of tumor recurrence were closely associated with clusterin expression (p <0.005 and <0.01, respectively). The recurrence-free and overall survival rates in patients with strong clusterin expression were significantly lower than in those with weak or no expression (p <0.05). Furthermore, multivariate analyses revealed that strong expression of clusterin was an independent predictor of tumor recurrence and overall survival (p <0.05). These findings suggest that the over expression of clusterin may be a useful prognostic parameter in patients with nonpapillary renal cell carcinoma after radical nephrectomy.

OVER EXPRESSION OF CLUSTERIN IS AN INDEPENDENT PROGNOSTIC FACTOR FOR NONPAPILLARY RENAL CELL CARCINOMA

Recent studies have revealed the powerful anti-apoptotic activity of clusterin in several types of malignant tumors. However, the significance of clusterin expression in human renal cell carcinoma has not been well characterized. We determined whether the expression level of clusterin in nonpapillary renal cell carcinoma may be used as predictor of tumor progression and prognosis. Total RNA was extracted from 93 nonpapillary renal cell carcinomas obtained at surgery and expression levels of clusterin messenger (m)RNA in these specimens were measured by Northern blot analysis. Findings were analyzed with respect to several clinicopathological factors. There were 48 cases (52%) with strong clusterin mRNA expression and 45 (48%) in which it was weak or nonexistent. The expression level of clusterin mRNA did not significantly correlate with patient gender, age, tumor grade or size, or histological cell type, whereas pathological stage and the incidence of tumor recurrence were closely associated with clusterin expression (p <0.005 and <0.01, respectively). The recurrence-free and overall survival rates in patients with strong clusterin expression were significantly lower than in those with weak or no expression (p <0.05). Furthermore, multivariate analyses revealed that strong expression of clusterin was an independent predictor of tumor recurrence and overall survival (p <0.05). These findings suggest that the over expression of clusterin may be a useful prognostic parameter in patients with nonpapillary renal cell carcinoma after radical nephrectomy.

Enhancement characteristics of papillary renal neoplasms revealed on triphasic helical CT of the kidneys.

The purpose of this study was to determine whether renal tumor enhancement or heterogeneity on triphasic helical CT scans is predictive of the papillary cell subtype or nuclear grade of renal cell carcinoma. We reviewed the CT scans of 90 consecutive patients with renal masses who had undergone triphasic renal helical CT before a complete or partial nephrectomy (12 with papillary renal cell carcinomas, 66 with nonpapillary renal cell carcinomas, and 12 with benign lesions). Three radiologists who were unaware of the patients' diagnoses retrospectively and independently measured the attenuation of each patient's tumor, abdominal aorta, and normal renal parenchyma on the scans obtained during all three phases. Ratios of tumor-to-aorta enhancement and tumor-to-normal renal parenchyma enhancement were calculated for both of the phases performed after contrast material had been administered. Tumor heterogeneity was calculated as the difference between the highest and lowest attenuation values divided by the value of the enhancement of the aorta. Values were correlated with cell type and nuclear grade found at surgical pathology. Low tumor-to-aorta enhancement and low tumor-to-normal renal parenchyma enhancement ratios on the vascular phase scans significantly correlated (p < 0.001) with papillary renal cell type carcinoma. Homogeneity and tumor-to-parenchyma enhancement ratios on the parenchymal phase scans also significantly correlated (p < 0.001) with papillary renal cell type carcinoma. Heterogeneity and tumor enhancement ratios did not correlate with the nuclear grade of the carcinoma. Papillary renal cell carcinomas are typically hypovascular and homogeneous. A high tumor-to-parenchyma enhancement ratio (> or = 25%) essentially excludes the possibility of a tumor being papillary renal cell carcinoma. A low tumor-to-aorta enhancement ratio or tumor-to-normal renal parenchyma enhancement ratio is more likely to indicate papillary renal cell carcinoma.

Proliferating cell nuclear antigen and epidermal growth factor receptor (EGFr) status in renal cell carcinoma patients with polysomy of chromosome 7

We investigated 40 cases of renal cell carcinoma (RCC) to study the polysomy 7 status in papillary and clear-cell types (nonpapillary RCC) and relationship with clinical, pathological, and biological features such as grade, stage, tumor proliferation rate (PCNA expression) and epidermal growth factor receptor (EGFr) expression and thereby to understand the prognostic significance of polysomy 7 and EGFr expression. In a prospective study, chromosome 7 copy number was analyzed in tumor cells by using fluorescence in situ hybridization (FISH) with an α-satellite DNA probe for chromosome 7. Both proliferating cell nuclear antigen (PCNA) and EGFr expression were examined in paraffin sections by immunostaining. The relationship between clinicopathological and clinicobiological parameters was evaluated by appropriate statistical methods. Polysomy 7 was present in 100% of papillary and 56.2% of clear-cell types RCC. In clear-cell RCC, in comparison with polysomy 7-dominant (D) category (20–50% polysomy-7 cells), polysomy 7–major (M) category (>50% polysomy 7 cells) was associated with higher tumor grade ( P = 0.05). Polysomy 7 was also correlated with stage of the disease ( P = 0.006). The PCNA index ranged between 12.8–89.6% and was comparatively high in high-grade tumors ( P = 0.001). The PCNA index was also correlated with polysomy 7 ( P = 0.002), and the association was stronger in tumors with polysomy M versus polysomy D category ( P = 0.02). The EGFr expression did not correlate with either grade, stage, PCNA, or polysomy 7. The correlation of polysomy 7 with less favorable prognostic factors such as higher tumor grade, stage, and higher proliferative index in the present study indicates that polysomy 7 might be used as a prognostic predictor in clear-cell RCC. Evaluation of clinical end points will confirm the prognostic potential of the genetic marker polysomy 7 in our study.

Similar regions of human chromosome 3 are eliminated from or retained in human/human and human/mouse microcell hybrids during tumor growth in severe combined immunodeficient (SCID) mice.

By passaging microcell hybrids (MCHs) containing human chromosome 3 (chr3) on A9 mouse fibrosarcoma background through severe combined immunodeficient (SCID) mice (elimination test), we have previously defined a 1-Mb-long common eliminated region 1 (CER1) at 3p21.3, a second eliminated region (ER2) at 3p21.1-p14 and a common retained region (CRR) at 3q26-qter. In the present work, chr3 was transferred by microcell fusion into the human nonpapillary renal cell carcinoma line KH39 that contained uniparentally disomic chr3. Four MCHs were generated. Compared with KH39, they developed fewer and smaller tumors, which grew after longer latency periods in SCID mice. The tumors were analyzed in comparison with corresponding MCHs by chr3 arm-specific painting, 19 fluorescent in situ hybridization (FISH) probes, and 27 polymorphic markers. Three MCHs that maintained the intact exogenous chr3 in vitro lost one 3p copy in all 11 tumors. Seven of 11 tumors lost the exogenous 3p, whereas four tumors contained mixed cell populations that lacked either the exogenous or one endogenous KH39 derived 3p. In one MCH the exogenous chr3 showed deletions within CER1 and ER2 already in vitro. It remained essentially unchanged in 8/9 derived tumors. The third, exogenous copy of the 3q26-q27 region (part of CRR) was retained in 16/20 tumors. It can be concluded that the human/human MCH-based elimination test identifies similar eliminated and retained regions on chr3 as the human/murine MCH-based test.

Genetic identification of bilateral primary or metastatic nonpapillary renal cell carcinoma.

To clarify the clonality of bilateral tumours by genetic analysis of bilateral renal cell carcinomas (RCCs) using the VHL gene, which is inactivated in approximately 60% of RCCs and which plays a causal role in the development of most cases of nonpapillary RCC. The study included 20 patients; seven had von Hippel-Lindau disease, three had papillary RCC and 10 had nonpapillary RCC. Paraffin-embedded blocks of tumour tissue were obtained from two of the three patients with papillary RCC and from nine of 10 with nonpapillary disease; all three exons of VHL were examined by direct sequencing. As reported previously, no VHL mutations were found in papillary tumours. However, in five of the nine nonpapillary cases, VHL mutations were identified in tumours on one or both sides. Three of the tumours had the same mutation on both sides, confirming a common origin. In the remaining two patients, the mutation status differed between the sides, confirming a bilateral primary origin. The former cases were characterized by a relatively large tumour on one side and multiple tumours on the other. In nonpapillary RCC multiplicity may suggest a metastatic origin. Such genetic information will be useful in treating and following patients with bilateral renal tumours.

Molecular cytogenetic analysis of 17 renal cancer cell lines: increased copy number at 5q31-33 in cell lines from nonpapillary carcinomas.

Comparative genomic hybridization (CGH) was used to screen for genomic imbalances in cell lines derived from 13 nonpapillary renal‐cell carcinomas (RCCs), two papillary RCCs, one renal squamous‐cell carcinoma, and one transitional‐cell carcinoma of the renal pelvis. Aberrations were found in all 17 lines. The most frequent changes in nonpapillary RCC cell lines were gains of 5q (85%), 7q (69%), 8q (69%) and 1q (54%) and losses of 3p (92%), 8p (77%), 4q (62%) and 14q (54%). High‐level gains (HLGs) were detected at 4q12, 5p, 5q23‐33, 7q22‐qter, 8q23‐24, 10q21‐qter, 12p and 12q13‐22. By means of fluorescence in situ hybridization (FISH) we narrowed the smallest common region involving 5q gains to the genomic segment between D5S642 and D5S673, and found that the HLG at 4q12 possibly involved amplifications of c‐kit and PDGFRA. Two papillary RCC cell lines showed gains of entire chromosomes 7, 12 and 17. The CGH data reported here should help to facilitate the choice of individual renal‐tumor cell lines for exploring target genes in regions of interest.

Expression of CD44 adhesion molecules in nonpapillary renal cell carcinoma and normal kidneys

Objectives. To analyze the relationship between the alterations in the expression of the CD44 gene in nonpapillary renal cell carcinoma (RCC) and several clinicopathologic factors. Methods. The expression of the CD44 gene in 10 human RCC cell lines, 60 nonpapillary RCC tumor samples, and 15 normal kidney samples was investigated by reverse-transcription polymerase chain reaction (RT-PCR) using a set of primers capable of amplifying all CD44 variant isoforms. The results were analyzed with respect to several clinicopathologic factors. Results. Nine of the 10 human RCC cell lines predominantly expressed the standard CD44 isoform (CD44s); CD44v10 was the major isoform in the 10th RCC cell line. The 15 normal kidney samples revealed the identical CD44 gene expression pattern; that is, CD44s, CD44v8-10, and CD44v10 were detectable in normal kidneys, and among them, CD44s was expressed most dominantly. In the 60 nonpapillary RCC samples, CD44s, CD44v8-10, and CD44v10 were the major isoforms in 46 (77%), 11 (18%), and 3 (5%) cases, respectively. Furthermore, the incidence of the predominant expression of CD44v8-10 in high-stage RCC was significantly higher than that in low-stage RCC. CD44s was more frequently expressed as a major isoform in clear cell RCC than in other histologic types of RCC. Conclusions. The results of this study show that the alternative splicing pattern of CD44 gene in RCC is different in each histologic type of RCC and suggest that CD44v8-10 upregulation in the progression of nonpapillary RCC is important.

Comparative allelotyping of the short arm of human chromosome 3 in epithelial tumors of four different types

Comparative allelotyping of the short arm of human chromosome 3 (3p) in four types of epithelial carcinomas was performed using an identical set of polymorphic markers. In total, 117 samples of non-papillary renal cell carcinoma (RCC), non-small cell lung carcinoma (NSCLC), carcinoma of uterine cervix (CC), and breast carcinoma (BC) were screened for loss of heterozygosity (LOH) with 10 di-, tri- and tetrameric markers covering nine bands of 3p. High LOH frequencies were detected in at least one locus: RCC (36/43, 84%), BC (20/26, 77%), NSCLC (16/24, 67%), and CC (15/24, 62%). Small interstitial deletions prevailed in BC and CC whereas large continuous and discontinuous deletions were mainly found in RCC and NSCLC. Different epithelial tumors displayed unique LOH profiles with partial overlaps in 3p26.1, 3p21.31, and 3p13. The overlap around D3S2409 (3p21.31) appeared common for RCC, BC and CC.