Ikeda R

Induction of centrosome amplification and chromosome instability in human bladder cancer cells by p53 mutation and cyclin E overexpression

Centrosome amplification frequently occurs in human cancers and is a major cause of chromosome instability (CIN). In mouse cells, centrosome amplification can be readily induced by loss or mutational inactivation of p53. In human cells, however, silencing of endogenous p53 alone does not induce centrosome amplification or CIN, although high degrees of correlation between p53 mutation and CIN/centrosome amplification in human cancer can be detected, suggesting the presence of additional regulatory mechanism(s) in human cells that ensures the numeral integrity of centrosomes and genomic integrity. Cyclin E, a regulatory subunit for CDK2 that plays a key role in centrosome duplication, frequently is overexpressed in human cancers. We found that cyclin E overexpression, together with loss of p53, efficiently induces centrosome amplification and CIN in human bladder cancer cells but not by either cyclin E overexpression or loss of p53 alone. We extended these findings to bladder cancer specimens and found that centrosome amplification is strongly correlated with concomitant occurrence of cyclin E overexpression and p53 inactivation but not with either cyclin E overexpression or p53 inactivation alone. Because cyclin E expression is strictly controlled in human cells compared with mouse cells, our findings suggest that this stringent regulation of cyclin E expression plays an additional role underlying numeral homeostasis of centrosomes in human cells and that deregulation of cyclin E expression, together with inactivation of p53, results in centrosome amplification.

Value of whole-body FDG PET in management of lung cancer

Abstract18F-fluorodeoxyglucose (FDG) PET imaging provides physiologic and metabolic information that characterizes lesions that are indeterminate by CT. FDG PET imaging is sensitive to the detection of lung cancer in patients who have indeterminate lesions on CT, whereas low grade malignancy such as bronchioloalveolar carcinoma and carcinoid may be negative on FDG PET. The specificity of PET imaging is less than its sensitivity because some inflammatory processes, such as active granulomatous infections, avidly accumulate FDG. This possibility should be kept in mind in the analysis of PET studies of glucose metabolism aimed at differentiating malignant from benign solitary pulmonary nodules. FDG uptake is considered to be a good marker of cell differentiation, proliferative potential, aggressiveness, and the grade of malignancy in patients with lung cancer. FDG PET accurately stages the distribution of lung cancer. Several studies have documented the increased accuracy of PET compared with CT in the evaluation of the hilar and mediastinal lymphnode status in patients with lung cancer. Whole-body PET studies detect metastatic disease that is unsuspected by conventional imaging. Management changes have been reported in up to 41% of patients on the basis of the results of whole-body studies. Whole-body FDG PET is also useful for the detection of recurrence. Several studies have indicated that the degree of FDG uptake in primary lung cancer can be used as an independent prognostic factor. Thus, whole-body FDG PET is clinically very useful in the management of lung cancer.

Centrosome Hyperamplification and Chromosomal Instability in Bladder Cancer

OBJECTIVE Chromosomal instability (CIN) is a common feature of malignant tumors. Centrosome hyperamplification (CH) occurs frequently in human cancers, and may be a contributing factor in CIN. In this study, we investigated the relationship between CH and CIN in bladder cancer. METHODS Clinical samples obtained by transurethral resection from 22 patients with bladder cancer were examined (histological grade G1, 5 cases; G2, 6 cases; G3, 11 cases). CH was evaluated by immunohistochemistry using anti-pericentrin antibody. CIN was evaluated by fluorescence in situ hybridization (FISH). FISH probes for pericentromeric regions of chromosomes 3, 7, and 17 were hybridized to touch preparations of nuclei from frozen tissues. We also analyzed the centrosome replication cycle of bladder cancer by laser scanning cytometry (LSC). RESULTS Of the 22 cases examined, 18 (81.8%) had centrosome hyperamplification: CH 0, 4 cases (18.1%); CH I, 5 cases (22.7%); CH II, 5 cases (22.7%); CH III, 8 cases (36.4%). The grade of CH was directly proportional to the histological grade (p=0.03, chi(2) test). LSC analysis showed that the centrosome replication cycle was well regulated in pathologically low-grade bladder cancer, which did not have chromosomal instability. In contrast, we found marked variability of centrosomes in pathologically high-grade bladder cancer, which had chromosomal instability. CH and CIN were both detected in pathologically high-grade tumors. The grade of CH was directly proportional to the CIN grade (p=0.0079, chi(2) test). CONCLUSION The results of the present study suggest that CH may be involved in CIN in bladder cancer.

Centrosome Hyperamplification and Chromosomal Damage after Exposure to Radiation

Objective: In order to elucidate the effects of radiation on centrosome hyperamplification (CH), we examined the centrosome duplication cycle in KK47 bladder cancer cells following irradiation. Methods: KK47 cells were irradiated with various doses of radiation and were examined for CH immunostaining for γ-tubulin. Results: Nearly all control cells contained one or two centrosomes, and mitotic cells displayed typical bipolar spindles. The centrosome replication cycle is well regulated in KK47. Twenty-four hours after 5-Gy irradiation, ∼80% of irradiated cells were arrested in G2 phase, and at 48 h after irradiation, 56.9% of cells contained more than two centrosomes. Laser scanning cytometry performed 48 h after irradiation showed the following two pathways: (1) unequal distribution of chromosomes to daughter cells, or (2) failure to undergo cytokinesis, resulting in polyploidy. With mitotic collection, M-phase cells with CH could be divided into G1 cells with micronuclei and polyploidal cells. Fluorescence in situ hybridization analysis showed clear signs of chromosomal instability (CIN) at 48 h after irradiation. The present study had two major findings: (1) continual duplication of centrosomes occurred in the cell cycle-arrested cells upon irradiation, leading to centrosome amplification; (2) cytokinesis failure was due to aberrant mitotic spindle formation caused by the presence of amplified centrosomes. Abnormal mitosis with amplified centrosomes was detected in the accumulating G2/M population after irradiation, showing that this amplification of centrosomes was not caused by failure to undergo cytokinesis, but rather that abnormal mitosis resulting from amplification of centrosomes leads to cytokinesis block. Conclusion: These results suggest that CH is a critical event leading to CIN following exposure to radiation.

P-glycoprotein expression is associated with FDG uptake and cell differentiation in patients with untreated lung cancer.

In vitro studies demonstrated that the accumulation of 2-deoxy-D-glucose was reduced in multidrug resistant cell lines. In animal study, it has been suggested that 2-[18F]fluoro-2-deoxy-D-glucose (FDG) may be a marker for multidrug resistance (MDR). The aim of this clinical study was to compare MDR characteristics by immunohistochemical assay with FDG uptake and investigate whether FDG is a marker for MDR in patients with untreated lung cancer. Forty-seven patients with 49 untreated lung cancers, who had undergone both preoperative FDG PET imaging and thoracotomy, were enrolled in this study. Before surgery, FDG PET was performed 40 min after injection, and standardized uptake values (SUVs) were obtained. Patients were classified into low-SUV (⩽5) and high-SUV (>5) groups. After surgery, the expression of P-glycoprotein (Pgp) was investigated by immunohistochemistry, and the lung cancer FDG uptake was analysed for possible association with Pgp expression. The strong intensity of Pgp immunoreactivity was seen only in the low-SUV group. The percentage of the Pgp positive area was significantly lower in the high-SUV group (21.7±13.4%) than in the low-SUV group (44.1±29.7%) (P = 0.015). In the high-SUV group, the percentage of Pgp positive area did not exceed 50%. In lung adenocarcinoma, the intensity of Pgp immunoreactivity and the percentage of Pgp positive area increased with degree of cell differentiation, while FDG uptake decreased with degree of cell differentiation. Bronchioloalveolar carcinoma, in particular, showed overexpression of Pgp and modest uptake of FDG. In conclusion, Pgp expression was found to be inversely related to FDG uptake in untreated lung cancer. Pgp expression correlated with the degree of cell differentiation in adenocarcinomas, whilst FDG uptake was inversely related to cell differentiation. FDG may be an in vivo marker for MDR in patients with untreated lung cancer.

Telomerase Activity in Giant Condyloma acuminatum

A 46-year-old male came to our hospital 1 month after noticing a 2-cm penile tumor. Since malignant findings such as atypical cells and mitosis were not observed in the frozen sections obtained at operation, the pathological diagnosis of this tumor was giant condyloma acuminatum. This tumor was analyzed by a telomeric repeat amplification protocol method, and telomerase activity was revealed. For comparison, a case of squamous cell carcinoma and a case of condyloma acuminatum were examined. Telomerase activity was observed in our case and in the case of squamous cell carcinoma. To our knowledge, this is the first case of telomerase activity in giant condyloma acuminatum ever reported. In addition to the histological examination, measurement of telomerase activity may provide valuable objective diagnostic information on evaluating the degree of malignancy of giant condyloma acuminatum and in obtaining a differential diagnosis between the benign and malignant.