肿瘤坏死因子相关诱导凋亡配体联合化疗药物治疗膀胱癌的研究

目的　探讨肿瘤坏死因子相关诱导凋亡配体 (TRAIL)治疗膀胱肿瘤的作用 ,以及与化疗药物的协同作用。方法　将T2 4及 5 63 7膀胱肿瘤细胞接种至 96孔培养板后分别加入浓度为1、10、10 0 μg/L的TRAIL ,0 .1、1.0、10 .0mg/L的阿霉素 (ADM )和丝裂霉素 (MMC) ,不同浓度的TRAIL、ADM、TRAIL和MMC ,噻唑蓝比色 (MTT)法分别检测肿瘤细胞的生存率。将膀胱肿瘤细胞接种至 12孔板 ,培育 2 4h后加入不同浓度的TRAIL、ADM、MMC、TRAIL联合ADM、MMC。用流式细胞术检测不同处理组肿瘤细胞的凋亡率和死亡率。结果　 10 0 μg/LTRAIL引起T2 4、5 63 7细胞的凋亡率分别为 2 0 .1%、45 .3 % ,与无药物组 1.1%、3 .5 %的凋亡率比较差异有非常显著性(P <0 .0 1)。单独运用 10mg/LMMC、ADM对T2 4、5 63 7的抑制率分别为 3 6.0 %、44 .1%、2 6.7%、3 0 .2 % ;而 10 0 μg/LTRAIL和 10mg/LMMC、ADM联合后对T2 4、5 63 7的抑制率分别达到 5 8.4%、73 .7%、90 .7%、88.2 % ,两者有明显的协同作用 (P <0 .0 5 )。结论　在体外实验中 ,TRAIL可通过诱导肿瘤细胞的凋亡而产生抗膀胱肿瘤的作用 ;TRAIL与化疗药物ADM、MMC有协同抗肿瘤作用     

Epidermal growth factor in urine from patients with bladder cancer

Epidermal growth factor (EGF), a mitogenic polypeptide with a molecular weight of 6000, is excreted in human urine in nanomolar quantities. Recently, some reports showed that urothelial neoplasm was related to the concentration of EGF in urine. In this study, EGF concentration in urine was measured by enzyme-linked immunosorbent assay (ELISA) in 207 samples from 112 male patients (30–90 years old, median 66.2) who had previously been treated for bladder cancer. Then, we tried to clarify the significance of urinary EGF as a marker for recurrence of bladder cancer in comparison with urine cytology. The samples were collected on occasional follow up cystoscopy. Urine from nine age-adjusted males without urological disease was also measured to obtain normal control values. In 123 samples from patients without tumors, EGF concentrations in urine decreased with age. In 84 samples obtained from patients with recurrent tumor, EGF concentrations were significantly lower than those in 123 samples from patients without tumors (P < 0.001) Furthermore, EGF concentrations in longitudinal samples collected the same patients during tumor recurrence and at the times when no tumor was detected were measured in 56 patients. EGF concentrations in the samples collected during tumor recurrence was significantly lower than that in specimens collected when there was no tumor (P < 0.01). There were no significant differences between the same samples collected during tumor recurrence with regard to tumor grade, stage shape and number of tumors. However, EGF concentration in urine from patients with carcinoma in situ (CIS) was lower than that in specimens from patients without CIS. These results indicate the usefulness of determining the EGF concentration as a marker for detecting bladder cancer recurrence. Urine cytology was also examined in the same series and findings were compared with those of urinary EGF. On cytology, class IV and V were considered positive, and on urinary EGF, less than l0 ng/mgCr were considered positive. Sensitivity was 25% for cytology and 57% for urinary EGF, while specificity was 98% and 66%, respectively. The predictive positive value was 0.88 and 0.53, respectively. With the combined use of urinary EGF and cytology, the sensitivity, specificity and predictive positive value were 68%, 64% and 0.92, respectively. In conclusion, urinary EGF seems to be a useful marker for detecting bladder cancer recurrence if performed in addition to cytology. Received: 8 April 1999 / Accepted: 3 January 2000     

三氧化二砷抑制人膀胱癌细胞BIU-87体外生长的实验研究

目的：观察三氧化二砷对人膀胱癌细胞的生长抑制作用,并探讨其机制。方法：采用ＭＴＴ法检测不同浓度的Ａｓ２Ｏ３对人膀胱细胞株ＢＩＵ＿８７的生长抑制率,原位末端转移酶标记技术检测细胞凋亡,ＳＡＢＣ免疫组织化分析ＢＩＵ－８７中ｂｃｌ－２的表达。结果：Ａｓ２Ｏ３可有铲地抑制ＢＩＵ＿８７细胞生长,具有时间、浓度依赖性特点;经药物作用后,膀胱癌凋亡细胞明显增多,凋亡率随作用时间的延长而增高,ＢＩＵ－８７细胞中的     

Paclitaxel and cisplatin as intravesical agents against non-muscle-invasive bladder cancer

OBJECTIVES

To investigate the effects of cisplatin and paclitaxel against human bladder cancer cells in vitro, and to obtain both pharmacokinetic and pharmacodynamic data after intravesical administration in mice.

MATERIALS AND METHODS

Six bladder cancer cell lines (J82, KU7, RT4, SW780, T24, UMUC3) were treated with various combined doses of both drugs and cell proliferation was evaluated 3 days later. In vivo, solutions of cisplatin and micellar paclitaxel were instilled transurethrally in female mice and pharmacokinetic data were acquired using high‐performance liquid chromatography‐mass spectrometry and atomic absorption methods. To obtain efficacy data, mice with orthotopic KU7‐luc tumours were administered cisplatin and/or micellar paclitaxel intravesically, and the tumour burden quantified using bioluminescence imaging.

RESULTS

In vitro, both cisplatin and paclitaxel potently decreased the proliferation of all cell lines tested, and in combination had an additive but not a synergistic effect. After intravesical instillation, mouse serum concentrations of cisplatin and paclitaxel were in the low microgram/millilitre range and bladder tissue concentrations achieved were 82 and 241 µg/g, respectively. Similar drug levels were reached using combined therapy. In vivo, all chemotherapeutic agents significantly inhibited bladder tumour growth, with the best results for combined therapy and micellar paclitaxel alone. However, there was toxicity in the combined treatment arm.

CONCLUSIONS

Both cisplatin and paclitaxel were absorbed at effective amounts into bladder tissues. As intravesical agents, paclitaxel had slightly stronger anticancer potency than cisplatin. Due to increased adverse events, caution should be exercised when combining both cisplatin and paclitaxel intravesically.     

干扰素结合化疗药物对膀胱癌细胞株BIU-87的抑制作用

在离体条件下，采用噻唑蓝（MTT）染色法，观察干扰素（IFN）与丝裂霉素C（MMC）、阿霉素（ADM）、噻替哌（TSPA）和羟基喜树碱（HCPT）等化疗药物联合应用对人类膀胱癌细胞株BIU－87的抑制作用。结果在连续联合用药组，浓度为25000u／ml的IFN与上述药物50％抑制率（ID50－1）剂量联合使用的抑制率分别为57．4％、58．6％、55．3％和64．3％；在序贯联合用药组，将MMC、ADM的ID50－2剂量作用于细胞24h后，再使用浓度为25000u／ml的IFN，24h后两者联合使用的抑制率分别为59．4％和54．3％。认为IFN与MMC等化疗药物联合应用能够取得较好的协同作用，为临床治疗浅表性膀胱肿瘤提供了新的思路。     

p53 and retinoblastoma pathways in bladder cancer

A majority of the aggressive, invasive bladder carcinomas have alterations in the p53 and retinoblastoma genes and pathways. Examination of the alterations in the molecules in these pathways that regulate the cell cycle and their effects on the prognosis of bladder cancer are areas of active research. While defects in the p53-Mdm2-p14 axis have been implicated in urothelial cancer, perturbations in the cyclin-dependent kinases and their inhibitors have also been extensively studied in this context. Genetic alterations of the retinoblastoma gene and aberrant post-translational modifications of its protein have also been incriminated in invasive bladder cancer. This article reviews the individual prognostic roles of alterations in these molecules in the context of bladder cancer. Additionally, we review findings from recent studies that are attempting to analyze these markers in combination in an effort to construct molecular panels that can serve as more robust outcome predictors. More importantly, alterations in these molecules are now becoming enticing targets for novel therapeutics. We also review some of these agents that can restore the tumor cells’ altered homeostatic mechanisms, thereby having potential in transitional cell carcinoma therapy. Future management of bladder cancer will employ validated marker panels for outcome prediction, and novel genetic and pharmacologic agents that will be able to target molecular alterations in individual tumors based on their respective profiles. A.P. Mitra and M. Birkhahn contributed equally to this paper.     

Use of nomograms as predictive tools in bladder cancer

Bladder cancer is a common genitourinary malignancy that demonstrates a great variation in risk of tumor recurrence and progression following treatment. The dramatic differences in clinical behavior dictate vastly differing treatments, which may range from simple surveillance to combination radical surgery with systemic chemotherapy. For non-muscle invasive bladder cancer prediction of the risk of recurrence and progression is necessary to assess the need for intravesical therapy and possible early cystectomy. In contrast, prediction of advanced disease response to primary treatment such as cystectomy and the response to systemic chemotherapy plays an important role in treatment assignment for patients with muscle invasive disease. To estimate these risk traditional risk grouping schemes such as the present TNM staging system has been used to guide patient treatment. More recently, improved prognostic tools such as nomograms have been developed to provide a more accurate assessment of outomes. Clinicians are enthusiastically working to utilize these statistical methods in bladder cancer. We summarize the current status of outcome predictive models for bladder cancer; and focus particularly on the ability of nomograms to predict disease recurrence, progression, and patient survival.     

膀胱癌特异性核基质蛋白-4在膀胱癌患者尿液中的表达及其临床意义

目的 探讨膀胱癌特异性核基质蛋白-4(BLCA-4)在膀胱癌患者早期诊断及术后监测肿瘤复发中的意义.方法 采用酶联免疫吸附试验(ELISA)法检测良性病变组、对照组和实验组患者术前及术后1、6个月和1年尿液中BLCA-4的表达.结果 实验组患者尿液中BLCA-4的含量中位数为39.48 A/μg protein,敏感性为93.33％,良性病变组和对照组分别为2.19 A/μg protein和0.965 A/μg protein,特异性为100％,2例复发患者术后1年复发时尿液中BLCA-4的含量均明显高于术后6个月时的含量.结论 BLCA-4作为一种新的膀胱肿瘤检测指标,有助于膀胱肿瘤的早期诊断及术后随访.     

Concomitant presence of bladder cancer and neurogenic bladder in a patient with HTLV-1 carrier: A case report

We describe a case of an HTLV-1 carrier who developed bladder cancer and neurogenic bladder. HTLV-1 is thought to alter host immune function and to contribute to the development of other malignancies. It is also sometimes reported that urinary symptoms precede pyramidal symptoms in patients with HAM. To our knowledge, concomitant presence of bladder cancer and neurogenic bladder in an HTLV-1 carrier has not been previously reported. This revised version was published online in August 2006 with corrections to the Cover Date.     

High expression of long noncoding RNA NORAD indicates a poor prognosis and promotes clinical progression and metastasis in bladder cancer

Purpose

To explore the function of NORAD in bladder cancer (BC), and to verify whether NORAD could be used as a biomarker to determine preoperative presence of progression and lymph node metastasis. To our knowledge, it is the first study investigating NORAD and its implications in BC.

Neoadjuvant and adjuvant chemotherapy in muscle-invasive bladder cancer

Context

The use of neoadjuvant and adjuvant chemotherapy in the treatment of muscle-invasive bladder cancer is still controversial.

Objective

To determine the optimal use of chemotherapy in the neoadjuvant and adjuvant settings in patients with advanced urothelial cell carcinoma. Bladder preservation is also discussed.

Evidence acquisition

A critical review of the published literature on chemotherapy for patients with locally advanced bladder cancer was performed.

Evidence synthesis

The presence of occult micrometastases at the time of radical cystectomy leads to both distant and local failure in patients with locally advanced transitional cell carcinoma of the bladder. Both neoadjuvant and adjuvant therapies have been evaluated in patients with locally advanced bladder cancer. Studies evaluating adjuvant chemotherapy have been limited by inadequate statistical power to detect meaningful clinical answers as well as by experimental arms utilizing inadequate chemotherapy.

Conclusions

The aggregate of available evidence suggests that neoadjuvant cisplatin-based combination chemotherapy should be considered as a standard of care for patients with muscle-invasive or locally advanced operable bladder cancer. In patients who are either unfit for or refuse radical cystectomy, neoadjuvant chemotherapy with or without radiation can render bladder preservation possible for patients who attain an excellent clinical response. With the introduction of new cytotoxic drugs, there is a need for well-designed studies to address the optimal utility of perioperative therapy in high-risk patients with bladder cancer.     

MicroRNA-93 promotes bladder cancer proliferation and invasion by targeting PEDF

Objective

MicroRNA-93 (miR-93) is upregulated in the urine of patients with bladder cancer (BC). Here, we investigated the role of miR-93 in BC progression and explored the underlying mechanism.

极低密度脂蛋白受体基因促进膀胱癌细胞迁移的作用及机制

目的:探讨极低密度脂蛋白受体(VLDLR)对人膀胱癌细胞迁移活性的影响及机制。方法:采用实时荧光定量反转录-聚合酶链反应(RT-qPCR)检测人源膀胱癌细胞(J82、UMUC3与T24)与正常尿路上皮细胞(SV-HUC-1)VLDLR的表达水平;采用小干扰RNA(siRNA)建立VLDLR敲低T24细胞模型,通过划痕实...     

Introduction of wild-type p53 gene downregulates the expression of H-ras gene and suppresses the growth of bladder cancer cells

Retroviral vectors were used to introduce the wild-type p53 gene into human bladder cancer cell lines BIU-87 and EJ, which express endogenous wt-p53 gene and have a mutation in H-ras gene. The expression of the exogenous wt-p53 gene in cells suppresses the growth of the bladder cancer cells in standard culture and in soft agar and blocks the cell cycle progression in G1. The BIU-87 and EJ cells developed tumors with average volumes of 6.53 cm³ and 6.61 cm³ in nude mice in 9 weeks after inoculation, while the cells transduced with wt-p53 gene failed to form tumors. The expression of H-ras gene in bladder cancer cells was reduced at mRNA level. These results suggest that the overexpression of the wt-p53 gene suppresses the expression of mutant H-ras gene and inhibits the tumor cell growth in vivo and in vitro.     

丁酸钠对膀胱癌细胞生长的影响

目的 观察丁酸钠对膀胱癌细胞生长的影响.方法 不同浓度的丁酸钠干预后,采用~3H-TdR掺入试验比较了膀胱癌BIU-87和E-J细胞的生长曲线变化,并采用流式细胞术研究了丁酸钠对膀胱癌细胞周期的影响.结果 1 mmol/L浓度组在各观察时间点均未显示出对E-J细胞的抑制;5、10mmol/L在各观察点显示出明显的生长抑制作用,各组之间差异有统计学意义(P<0.01);随着丁酸钠浓度的升高,越来越多的BIU-87和E-J细胞细胞被阻滞在G_0/G_1期.结论 丁酸钠对肿瘤细胞增殖的抑制作用具有浓度、时间依赖性,这种作用是通过G_0/G_1期阻滞实现的.     

Real-time cancer cell tracking by bioluminescence in a preclinical model of human bladder cancer growth and metastasis

Background

Bladder cancer is the fifth most common malignancy in the Western world and the second most frequently diagnosed genitourinary tumor. In the majority of cases, death from bladder cancer results from metastatic disease. Understanding the multistep process of carcinogenesis and metastasis in urothelial cancers is pivotal to the development of new therapeutic strategies. Molecular imaging of cancer growth and metastasis in preclinical models provides the essential link between cell-based experiments and clinical translation.

Objective

Develop preclinical models for sensitive bladder cancer cell tracking during tumor progression and metastasis.

Design, setting, and participants

A human transitional cell carcinoma UM-UC-3 cell line was generated that stably expresses luciferase 2 (UM-UC-3luc2), a mammalian codon-optimized firefly luciferase with superior expression. Preclinical models were developed with human UM-UC-3luc2 cells xenografted into the bladder (orthotopic model with metastases) or inoculated into the left cardiac ventricle (bone metastasis model) of immunocompromised mice.

Measurements

Noninvasive, sensitive bioluminescent imaging of human firefly luciferase 2-positive bladder cancer in mice using the IVIS100 imaging system.

Results and limitations

In the orthotopic model (intravesical inoculation), tumor growth could be followed directly after inoculation of UM-UC-3luc2 cells. Importantly, micrometastatic lesions originating from orthotopically implanted cancer cells could be detected in the locoregional lymph nodes and in distant organs. In addition, the superior bioluminescent indicator firefly luciferase 2 allows the detection and monitoring of micrometastatic lesions in real time after intracardiac inoculation of human bladder cancer cells in mice. The main disadvantage is the lack of T-cell immunity in the preclinical models.

Conclusions

The new bioluminescence-based preclinical bladder cancer models enable superior, noninvasive, and real-time tracking of cancer cells, tumor progression, and micrometastasis. Because of the significant improvement in detection of small cell numbers, the presented models are ideally suited for functional studies dealing with minimal residual disease as well as real-time imaging of drug response.     

Temsirolimus improves cytotoxic efficacy of cisplatin and gemcitabine against urinary bladder cancer cell lines

ObjectivesTo analyze the cytotoxic action of temsirolimus using 3 established human bladder cancer cell lines and to assess whether temsirolimus potentiates the anticancer activity of gemcitabine and cisplatin.MethodsTemsirolimus (500, 1,000, 2,000, and 4,000 nM), in isolation, and combined with gemcitabine (100 nM) and cisplatin (2.5 µg/ml), was given to 5637, T24, and HT1376 bladder cancer cell lines. Cell proliferation, autophagy, early apoptosis, and cell cycle distribution were analyzed after a 72-hour period. The expression of mammalian target of rapamycin baseline, Akt, and their phosphorylated forms, before and after treatment with temsirolimus, was evaluated by immunoblotting.ResultsTemsirolimus slightly decreased the bladder cancer cell proliferation in all 3 cell lines. No significant differences in the expression of mammalian target of rapamycin, Akt, and their phosphorylated forms because of temsirolimus exposure were found in the 3 cell lines. As part of a combined regime along with gemcitabine, and especially with cisplatin, there was a more pronounced antiproliferative effect. This pattern of response was similar to the other parameters analyzed (increased autophagy and apoptosis). Also, in the combined regime, an enhanced cell cycle arrest in the G0/G1 phase was observed. The non–muscle invasive 5637 bladder cancer cell line was most sensitive to both combinations.ConclusionsTemsirolimus makes a moderate contribution in terms of cell proliferation, apoptosis, and autophagy. However, it does potentiate the activity of gemcitabine and particularly cisplatin. Therefore, cisplatin- or gemcitabine-based chemotherapy regimen used in combination with temsirolimus to treat bladder cancer represents a novel and valuable treatment option, which should be tested for future studies in urinary bladder xenograft models.     

Establishment and characterization of a new cell line from human bladder cancer (JMSU1)

A new human bladder cancer cell line designated JMSU1 has been established from malignant ascitic fluid of a 75-year-old Japanese man with bladder cancer, and maintained in culture for more than 7 years and over 240 passages. Inverted phase-contrast microscopy revealed that JMSU1 was composed of morphologically distinct cells (polygonal to spindle-shaped cells), showing morphological heterogeneity in vitro. Histological examination of xenografts showed poorly differentiated transitional cell carcinoma, resembling the original tumor. Immunohistochemical staining for cytokeratin and electron microscopic examination suggested that JMSU1 was of epithelial origin. Chromosome analysis gave a modal number of 69 with no Y chromosome. Isozyme analysis (LDH, G6PD, and NP) showed the mobility pattern of human type B. DNA fingerprint analysis demonstrated that there was no cross-culture contamination of JMSU1 during the passages. In conclusion, a newly established and well-characterized cell line, JMSU1, offers promising material for the investigation of the biological properties of bladder cancer.