Meloxicam in the treatment of in vitro and in vivo models of urinary bladder cancer

To assess the efficacy of meloxicam, a non-steroidal anti-inflammatory drug (NSAID), on three human urinary bladder-cancer cell lines (HT1376, T24 and 5637) and on mice urinary bladder cancer chemically induced by N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN). The in vitro effects of meloxicam were assessed by optical microscopy, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method, flow cytometry and comet assay. In vivo, Hsd:ICR male mice were exposed to BBN in drinking water, over the course of 12 weeks. Subsequently, animals were treated with meloxicam by intraperitoneal route, for 6 consecutively weeks. Tumour development was evaluated by haematoxylin and eosin staining. Renal and hepatic functions, interleucin-6 (IL-6), C-reactive protein (CRP) and tumour necrosis factor (TNFα) were also evaluated. In vitro, meloxicam induced a significant (P < 0.05) decrease of cell proliferation. A significant (P < 0.05) cell cycle arrest on G₀/G₁ phase was also detected in all the cell lines, with a slight but significant increase of sub-G₀/G₁ fraction on T24 (P = 0.006) and 5637 (P < 0.001) cells. Also a significant (P < 0.05) increase in DNA damage was found on meloxicam-treated cells. In vivo, the incidence of pre-neoplastic lesions induced by BBN was not affected by meloxicam treatment. However, although not statistically significant, the development of neoplastic lesions was inhibited by meloxicam treatment without significant alterations of renal or hepatic parameters. Meloxicam is effective on in vitro and in vivo models of urinary bladder cancer. These findings support that meloxicam deserves more attention on urinary bladder cancer study.     

Cyclooxygenase inhibitors: Scope of their use and development in cancer chemotherapy

The traditional nonsteroidal anti‐inflammatory drugs (NSAIDs) exert their effect by inhibition of cyclooxygenase‐1 (COX‐1) as well as COX‐2 enzymes. As COX‐1 is responsible for maintaining normal biological functions, the nonselective inhibition of these enzymes caused side effects including gastrointestinal (GI) problems. Recently developed selective COX‐2 inhibitors could reduce these adverse effects, but the evidence of cardiovascular side effects including an increased risk of myocardial infarction began to emerge, and some of the COX‐2 inhibitors were eventually withdrawn from the market and this led to the downfall of this research. So, the discovery of novel COX‐2 inhibitors with their safety profile became the biggest challenge in pharmaceutical research. However, recent mechanistic and clinical studies revolutionized this area by indicating the fact that COX‐2 is involved in apoptosis resistance, angiogenesis, and tumor progression. Epidemiological data suggest that selective COX‐2 inhibitors might prevent the development of cancers. Moreover, COX‐2 is found to be overexpressed in many cancers thus making it an attractive therapeutic target for the prevention and treatment of a number of malignancies. The purpose of this review is to focus on the medicinal chemistry aspects of COX‐2 inhibitors in cancer chemotherapy and recent reports on these inhibitors as anticancer agents. We attempted to cover only the COX inhibitors that showed anticancer activity, although a number of potent COX‐2 inhibitors have been reported without their anticancer effects. Furthermore, structure–activity relationships (SAR) of different classes of compounds for COX‐2 inhibition as well as anticancer activity, and their future applications are discussed. © 2009 Wiley Periodicals, Inc. Med Res Rev, 31, No. 2, 161–201, 2011     

膀胱癌术后尿流改道与膀胱替代的现状与进展

根治性全膀胱切除是治疗肌层浸润性膀胱癌的金标准,但膀胱全切术后尿流改道方式的选择尚无统一标准。尿流改道术主要包括不可控尿流改道术、可控性尿流改道术、原位新膀胱术三种方式。在条件允许的情况下,应将原位新膀胱作为膀胱替代的首选方式。而乙状结肠原位膀胱更接近自然膀胱的功能,是一种理想的尿流改道方式。对于肿瘤侵犯尿道或尿道狭窄不能经尿道排尿的患者,乙状结肠直肠膀胱术是越来越被广泛接受的可控性尿流改道方式。Bricker回肠膀胱术因手术简单、并发症少,仍被广泛应用。而输尿管皮肤造口术则特别适用于年龄大、体质差、耐受力低、不能承受复杂手术的患者。组织工程技术为膀胱替代带来了新的希望,组织工程膀胱的一系列基础研究显示了广阔临床应用前景。     

Cyclooxygenase-2 dependent and independent antitumor effects induced by celecoxib in urinary bladder cancer cells

Transitional cell carcinoma of the urinary bladder is the second most common genitourinary malignancy in people in the United States. Cyclooxygenase-2 (COX-2) is overexpressed in bladder cancer. COX-2 inhibitors have had antitumor activity against bladder cancer, but the mechanisms of action are unclear. Clinically relevant concentrations of COX-2 inhibitors fail to inhibit proliferation in standard in vitro assays. In pilot experiments, different culture conditions [standard monolayer, modified monolayer, soft agar, collagen, and poly(2-hydroxyethyl methacrylate)-coated plates] were assessed to determine conditions suitable for the study of COX inhibitor growth-inhibitory effects. This was followed by studies of the effects of clinically relevant concentrations of a selective COX-2 inhibitor (celecoxib) on urinary bladder cancer cell lines (HT1376, TCCSUP, and UMUC3). Celecoxib (相似文献   

尿膀胱癌抗原、透明质酸和存活素联合检测对膀胱癌的诊断运用探讨

目的探讨在对膀胱癌的诊断过程中,尿膀胱癌抗原、透明质酸和存活素的应用价值。方法膀胱癌患者74例,泌尿系统良性疾病患者20例,健康志愿者标本10例分别行膀胱癌抗原(UBC)、透明质酸(HA)、存活素(Sur-vivin)和尿脱落细胞学(UC)检查。结果 UBC、HA、Survivin和尿脱落细胞学的检测有利于膀胱癌的诊断,且UBC、HA、Survivin三者的有效性优于尿脱落细胞学的检测。UBC、HA、Survivin在检测膀胱癌的过程中,三者的敏感性和准确性高于尿脱落细胞学。UBC、HA、Survivin在肿瘤的各分级(G1、G2、G3)和各分期(pTa、pT1、≥pT2)中的敏感性明显高于尿脱落细胞学。结论尿膀胱癌抗原、透明质酸和存活素在诊断膀胱癌过程中应用价值优于尿脱落细胞学。     

Treatment of bladder cancer cells in vitro and in vivo with 2-5A antisense telomerase RNA

Bladder cancer is the most common malignant tumor of the urinary tract. Novel treatment approaches are essential because of the failure of current treatment options to cure a high percentage of patients. Telomerase, a ribonucleoprotein, is detected in almost all bladder cancer, but not in normal bladder tissues. Therefore, telomerase is expected to be a very promising candidate for targeted therapy of bladder cancer. In this study, we synthesized a 19-mer antisense oligonucleotide against the RNA component of human telomerase (hTR) linked to a 2-5A molecule (2-5A-anti-hTR) and investigated its antitumor effect against bladder cancer cells. The 2-5A antisense strategy relies on the recruitment and activation of RNase L at the site of targeted RNA sequence. Here we demonstrate that treatment with 2-5A-anti-hTR reduced the viability of seven bladder cancer cell lines (UM-UC-2, UM-UC-3, UM-UC-6, UM-UC-9, UM-UC-14, RT4 and T24) expressing telomerase activity to 21-55% within 4 days. The cytotoxicity was mainly due to induction of caspase-dependent apoptosis. In contrast, normal fibroblast WI38 cells lacking telomerase activity were resistant to the treatment. Furthermore, treatment of subcutaneous UM-UC-2 tumors in nude mice with 2-5A-anti-hTR significantly suppressed the tumor growth through induction of apoptosis (P < 0.001). These findings may offer a strong support to the feasibility of the 2-5A-anti-hTR treatment for human bladder cancer.     

Current approaches to treatment of urinary bladder cancer

The study included 30 patients with surface cancer of the urinary bladder stage TA-T1 G1-3. As the first step of the treatment, all the patients were operated with removal of the tumor then the patients were randomized to postoperative intravesicular immunotherapy with ronkoleucine in single doses 500,000 IU (15 patients of group 1) or 1,000,000 IU (15 patients of group 2). It was found that group 2 patients had recurrences much less frequently (26.7 vs 66.7%, respectively). With higher degrees of differentiation of the tumor cells recurrences occurred more frequently in both groups. Group 2 patients developed recurrences significantly less frequently in G1 and G2 (22.2%). In G3 all the patients had recurrences. Intravesicular administration of ronkoleukine raised absolute number of CD3 and CD4 subpopulations during the treatment and after it as well as raised concentration of TNF. The levels of the latter in the urine rose after the end of each immunoprophylaxis course. Intravesicular use of ronkoleukine entailed no specific toxic reactions. Thus, intravesicular prophylactic immunotherapy of recurrent surface cancer of the urinary bladder with ronkoleukine in a single dose 1,000,000 IU is effective prevention in patients with high (G1) and moderate (G2) grade of tumor cell differentiation. The single dose 500,000 IU is uneffective. A rise in subpopulations CD3, CD4 and TNF cytokine in the urine evidences for systemic activation of the immunity.     

In vivo three-dimensional sonographic measurement of organ volume: validation in the urinary bladder.

The purpose of this study was to assess the accuracy of in vivo measurement of organ volume using 3DUS and compare the results to 2D sonographic methods using the urinary bladder as the target organ and voided urine volume for validation. Fifty normal volunteers were studied. 2D volume measurements were based on length, width, and depth data and assumed a regular geometric model. 3D volume measurements were based on masked slices with the voxels integrated over the entire bladder. Voided urine volumes ranged from 35 ml to 701 ml. Residual urine volume was present in 48% of the subjects and ranged from 1% to 14% of the voided volume. 2D volume estimates for all 50 subjects had a mean absolute value of the error of 27.5% +/- 17.8%. 3D volume measurements had a mean absolute value of the error of 4.3% +/- 3.7% (transverse) and 5.6% +/- 3.8% (longitudinal). 3DUS provided more accurate volume measurements than 2DUS, particularly for irregularly shaped organs.     

联合检测尿膀胱癌抗原和survivin诊断膀胱癌的临床应用

目的临床评价联合检测尿液中尿膀胱癌抗原（urinary bladder cancer antigen,UBC）和survivin基因诊断膀胱癌的临床应用价值。方法对64例膀胱癌患者、20例泌尿系其他良性疾病患者,在膀胱镜检查之前留尿将尿样分为3份,分别进行UBC、survivin和脱落细胞检测,分析比较三种方法诊断膀胱癌的临床应用价值。结果 UBC和survivin诊断膀胱癌的敏感度分别为85.9%（55/64）和93.8%（60/64）,与脱落细胞学（40.6%）比较,差异有统计学意义（P〈0.01〉,三种方法诊断膀胱癌的特异度分别为85.0%（17/20）、95%（19/20）和95%（19/20）。各分级和分期UBC和survivin诊断膀胱癌的敏感度均高于尿脱落细胞学检查;UBC值和survivin检测的敏感度在各分级和分期中差异无统计学意义（P〉0.05）;而尿脱落细胞学检查,肿瘤的分级越高,其敏感度越高（P〈0.01）,各分期之间差异无统计学意义（P〉0.05）。联合运用UBC和survivin,敏感度和特异度均达到100%。结论尿液中的UBC和survivin是早期诊断膀胱癌较好的肿瘤标志物,联合检测能提高诊断的敏感度和特异度。     

Ultrasonographic findings in gastric cancer: in vitro and in vivo studies

Ultrasonography was performed in 45 cases of gastric cancer. Specimens from all 45 cases of gastric cancer were subjects to ultrasonographic study by the water immersion method for comparison with histology. In 32 of these 45 cases in vivo ultrasonographic evaluation was performed prospectively. The overall accuracy rates for the diagnosis of the depth of cancerous invasion were almost 80% in both in vitro and in vivo studies. In vivo ultrasonographic findings agreed well with those from the specimen studies. Ultrasonography was considered to be useful in the diagnosis of gastric malignancies.     

Adenovirus-mediated gene therapy for bladder cancer: efficient gene delivery to normal and malignant human urothelial cells in vitro and ex vivo

Existing local therapies for superficial transitional cell carcinoma (TCC) of the bladder have limited success in preventing progression to life-threatening, muscle-invasive disease, and novel therapies are needed. Recent studies have raised doubts concerning the feasibility of adenovirus-mediated gene therapy for bladder cancer. We have therefore investigated adenoviral transduction of normal and malignant human urothelial cells, both as primary cultures and in intact epithelium.All 15 primary normal human urothelial cell lines tested were transduced in vitro by Adv-cmv-beta-gal at high efficiency, and better than most human TCC cell lines. Eight primary human TCC explants were also successfully transduced. In contrast, in intact normal urothelium, transduction efficiency was lower, and occurred only in superficial epithelial layers. Expression of the hCAR adenovirus receptor, however, occurred throughout the full thickness of urothelium. Transduction of human TCC biopsy specimens was at least as efficient as intact normal urothelium.We demonstrate for the first time that adenoviral transduction of both normal and malignant human urothelial cells is feasible. A physical barrier, rather than hCAR status, may be the main determinant of transduction of intact epithelium. Clinical trials of adenovirus-mediated gene therapy for superficial bladder cancer are warranted.     

Palliative ileal conduit in inoperable cancer of the urinary bladder

A retrospective study is presented of 19 patients with inoperable urinary bladder carcinoma in which urinary diversion was performed by ileal conduit without cystectomy. The postoperative mortality rate was 10.5% and the incidence of complications was 16%. The average time of survival was 5.1 months and varied from between one and 23 months. According to the literature these results are no better than those in patients undergoing no therapy at all. It is, thus, our opinion that palliative ileal conduit should not be considered a suitable method for urinary diversion in patients with advanced carcinoma of the urinary bladder.     

Factor VIII inhibitors: von Willebrand factor makes a difference in vitro and in vivo

Summary. Background: The important association between von Willebrand factor (VWF) and factor VIII (FVIII) has been investigated for decades, but the effect of VWF on the reactivity of FVIII inhibitory antibodies, referred to as inhibitors, is still controversial. Objective: To investigate the interaction among VWF, FVIII and FVIII inhibitory antibodies. Methods: Three sources of inhibitors were used for in vitro studies, including the plasma from immunized VWF^nullFVIII^null mice, purified plasma IgG from human inhibitor patients, or human monoclonal antibody from inhibitor patients’ B‐cell clones. Inhibitors were incubated with recombinant human FVIII (rhFVIII) either with or without VWF. The remaining FVIII activity was determined by chromogenic assay and inhibitor titers were determined. For in vivo studies, inhibitors and rhFVIII were infused into FVIII^null or VWF^nullFVIII^null mice followed by a tail clip survival test. Results: VWF has a dose‐dependent protective effect on FVIII, limiting inhibitor inactivation of FVIII in both mouse and human samples. A preformed complex of VWF with FVIII provides more effective protection from inhibitors than competitive binding of antibodies and VWF to FVIII. The protective effect of VWF against FVIII inactivation by inhibitors was further confirmed in vivo by infusing inhibitors and FVIII into FVIII^null or VWF^nullFVIII^null mice followed by a tail clip survival test. Conclusion: Our results demonstrate that VWF exerts a protective effect, reducing inhibitor inactivation of FVIII, both in vitro and in vivo.     

Enantiomers of oxybutynin: in vitro pharmacological characterization at M1, M2 and M3 muscarinic receptors and in vivo effects on urinary bladder contraction, mydriasis and salivary secretion in guinea pigs

The major side effects of racemic oxybutynin (OXY), which is used in the treatment of urinary incontinence are dry mouth (xerostomia) and blurred vision (mydriasis). Highly purified enantiomers of OXY [(R)OXY, (S)OXY] were compared with the racemate both in vitro in functional studies and in vivo in guinea pigs to evaluate their pharmacological action relative to their adverse effects. The affinity of (R)OXY and (S)OXY for different muscarinic receptor subtypes was determined using field stimulated rabbit vas deferens (M1) and guinea pig atria (M2) or bladder (M3) strips. Stereoselective antimuscarinic effects [(R)OXY greater than or equal to (R/S) OXY much greater than (S)OXY] were evident at all three receptor subtypes; the isomeric ratio [(S)OXY/(R)OXY] ranged from 12 to 88. Both (R)OXY and (R/S)OXY were slightly more selective (2-4-fold, P less than .01) for M1 and M3 relative to M2 muscarinic receptors. Stereoselectivity was also evident in vivo for volume-induced urinary bladder contractions as measured by cystometrogram parameters [(S)OXY/(R)OXY approximately 21], mydriasis [(S)OXY/(R)OXY approximately 136] and salivary gland secretory responses [(S)OXY/(R)OXY approximately 30]. The absolute potencies of (R)OXY or (R/S)OXY for mydriasis and salivation were similar to those for inhibition of intravesical bladder pressure. Also, (R)OXY and (R/S)OXY equipotently antagonized cholinergic-mediated CNS effects in mice. Collectively, the data suggest that the activity of (R/S)OXY resides predominantly in the (R)-enantiomer. However, it appears that (R)OXY may offer no significant pharmacological advantage over (R/S)OXY in terms of its principal therapeutic and side effect profile.     

Phosphodiesterase type 4 inhibitors cause proinflammatory effects in vivo

Phosphodiesterase type 4 (PDE(4)) inhibitors are currently being evaluated as potential therapies for inflammatory airway diseases. However, this class of compounds has been shown to cause an arteritis/vasculitis of unknown etiology in rats and cynomolgus monkeys. Studies in rodents have demonstrated the anti-inflammatory effects of PDE(4) inhibitors on lipopolysaccharide (LPS)-induced airway inflammation. The aim of this work was to assess the direct effects of PDE(4) inhibitors on inflammatory cells and cytokine levels in the lung in relation to therapeutic effects. The effects of the PDE(4) inhibitors 3-cyclo-propylmethoxy-4-difluoromethoxy-N-[3,5-di-chloropyrid-4-yl]-benzamide (roflumilast) and 3-(cyclopentyloxy)-N-(3,5-dichloro-4-pyridyl)-4-methoxybenzamide (piclamilast) were assessed in vivo, using BALB/c mice, and in vitro, in unstimulated human endothelial and epithelial cell lines. In BALB/c mice, LPS challenge caused an increase in neutrophils in bronchoalveolar lavage (BAL) and lung tissue and BAL tumor necrosis factor-alpha levels, which were inhibited by treatment with either roflumilast or piclamilast (30-100 mg/kg subcutaneously). However, roflumilast and piclamilast alone (100 mg/kg) caused a significant increase in plasma and lung tissue keratinocyte-derived chemokine (KC) levels, and lung tissue neutrophils. In vitro, both piclamilast and roflumilast caused an increase in interleukin (IL)-8 release from human umbilical vein endothelial cells but not BEAS-2B cells, suggesting that one source of the increased KC may be endothelial cells. At doses that antagonized an LPS-induced inflammatory response, the PDE(4) inhibitors possessed proinflammatory activities in the lung that may limit their therapeutic potential. The proinflammatory cytokines KC and IL-8 therefore may provide surrogate biomarkers, both in preclinical animal models and in the clinic, to assess potential proinflammatory effects of this class of compounds.     

尿膀胱癌抗原、核基质蛋白22和透明质酸诊断膀胱癌的临床价值

目的比较应用尿膀胱癌抗原(UBC)、核基质蛋白22(NMP22)以及透明质酸(HA)诊断膀胱癌的应用价值。方法 70例不同分级和分期膀胱癌患者、23例泌尿系统良性疾病患者留尿分别进行UBC、NMP22、HA以及尿脱落细胞学检测,比较4种方法诊断膀胱癌的价值。结果 UBC、NMP22和HA诊断膀胱癌的敏感性分别为84.3%(59/70)、81.4%(57/70)、88.6%(62/70),与脱落细胞学38.6%(27/70)比较,差异有统计学意义(P<0.05)。4种方法诊断膀胱癌的特异性分别为82.6%(19/23)、65.2%(15/23)、78.2%(18/23)、95.7%(22/23)。各分级和分期UBC、NMP22以及HA诊断膀胱癌的敏感性均高于尿脱落细胞学检查。UBC、NMP22值各分级和分期比较差异无统计学意义(P>0.05)。HA检测值G2、G3组均明显高于G1组(P<0.05),但G2、G3组间比较差异无统计学意义(P>0.05);各分期之间比较差异无统计学意义(P>0.05)。尿细胞学检查,肿瘤分级越高,敏感性越高(P<0.05),各分期之间比较差异无统计学意义(P>0.05)。结论尿UB...     

P27kipl、CyclinD1在膀胱癌中的表达及其意义

目的:通过检测P27kipl与CyclinD1在膀胱癌中的表达,以探讨两者在膀胱癌发生发展中的作用及其与临床病理特征的关系.方法:对80例标本进行SP免疫组织化学方法检测P27kipl、CyclinD1的表达水平,其中正常组织28例,膀胱癌组织52例.结果:P27kipl、CyclinD1在正常膀胱组织中的表达率和在膀胱癌组织中的表达差异均具有显著性(P＜0.01).P27kipl蛋白在低分化组中的表达水平显著低于高分化组(P＜0.01)和中分化组(P＜0.01),CyclinD1表达水平在低分化组中显著高于高分化组(P＜0.01),在中、低分化组间差异无显著性(P＞0.05).P27kipl的表达与临床分期无关,而CyclinD1的表达与临床分期相关.P27kipl与CyclinD1的相互关系呈负相关.结论:P27kipl蛋白的低表达或缺失在膀胱癌的发生中起重要作用,是膀胱癌发生中的早期分子事件.CyclinD1的过表达参与了膀胱癌的发生,并与其发展密切相关,可作为评价预后的指标之一.膀胱癌的发生发展与P27kipl、CyclinD1蛋白的异常表达密切相关,联合测定P27kipl、CyclinD1蛋白的表达对膀胱癌的诊断、治疗有一定的指导意义,可作为膀胱癌诊断和预后评价的重要参数.