Hyperprogression after anti-programmed death-1 therapy in a patient with urothelial bladder carcinoma:A case report

BACKGROUND Immune checkpoint inhibitors,including programmed death-ligand 1(PD-L1)and programmed death-1(PD-1)have recently been approved to treat locally advanced and metastatic urothelial carcinoma(UC).However,some patients experience rapid tumor progression rather than any clinical benefit from anti-PDL1/PD-1 therapy.CASE SUMMARY A 73-year-old woman with bladder UC showed the progression of multiple metastases after surgery and chemotherapy for over 12 mo.The patient could not tolerate further chemotherapy.Next-generation sequencing was performed,and the results indicated that the tumor mutational burden was 6.4 mutations/Mb.The patient received the anti-PD-L1 agent toripalimab combined with albuminbound paclitaxel.Compared with the baseline staging before immunotherapy,the patient had a treatment failure time of<2 mo,an increase in tumor burden of>50%,and a>2-fold increase in progression,indicating hyperprogression.CONCLUSION Selecting patients most likely to respond to treatment with immunotherapeutic agents remains challenging.For older patients with advanced UC who have already exhausted multi-line chemotherapy options,immunotherapy should be used prudently if no effective biomarker is available.Further studies are required to clarify the causes and mechanisms of hyperprogression.

Complete response to trastuzumab and chemotherapy in recurrent urothelial bladder carcinoma with HER2 gene amplification: A case report

BACKGROUND Targeted treatments may greatly affect the natural history of urothelial carcinoma based on their pharmacokinetics. A phase II trial has explored the combination of cytotoxic chemotherapy with the anti-HER-2 monoclonal antibody trastuzumab in selected patients with metastatic bladder cancer, but it failed.CASE SUMMARY Here, we report a case of recurrent urothelial bladder carcinoma(UBC) in a patient who has undergone three operations, and further illuminate its diagnosis and treatment. The diagnosis of UBC was rendered according to the pathological indices. Next-generation sequencing on formalin fixed paraffin-embedded(FFPE)tissue was also performed and suggested HER2 gene amplification in the FFPE tissue. Based on HER2 gene amplification in FFPE, the patient was treated with chemotherapy in combination with trastuzumab after his third surgery.Fortunately, the patient got a clinically complete remission to trastuzumab for 34 mo.CONCLUSION There is not enough clinical evidence for incorporating trastuzumab in routine treatment of UBC. This case hinted that recurrent UBC patients with HER2 gene amplification may benefit from targeted trastuzumab. Further studies are needed to further investigate the status of HER2 gene and better determine trastuzumab in the management of UBC.

THE INHIBITORY EFFECT OF MELATONIN ON THE GROWTH OF HUMAN BLADDER CARCINOMA T24 CELL LINE

Objective To study the inhibitory effects of melatonin and its inhibitory mechanism on the growth of human bladder carcinoma T24. Methods The inhibitory effects of melatonin with various concentrations on the human bladder carcinoma T24 lines in vitro were determined by MTT assay. The mechanism of the inhibition was observed by flow cytometry (FCM) and transmission electron microscopy (TEM). Results The 30% inhibition concentration (IC 30) value was 0.71 mmol·L -1 and the 50% inhibition concentration (IC 50) value was 1.20 mmol·L -1. The population doubling time of T24 cells treated with melatonin at 0.71 mmol·L -1 was 43.2 hours, which was significant different from that of 34.6 hours of the control group. Using FCM, we found that the cell percentage increased during the G 1 phase, but decreased during the S stage. The degenerated ultra-structure of the cell treated with melatonin was also observed by TEM. Conclusion The results suggest that melatonin can inhibit the growth of human bladder carcinoma T24. The inhibitory effects of melatonin might be the prolonging of the staging from G 1 to S in the cell cycle.

EXPRESSION OF A MUTANT hTERT IN HUMAN BLADDER CARCINOMA CELL LINE T24 AND ITS CLINICAL SIGNIFICANCE

To construct a mutant pEGFP- hTERTexpression vector, to observe its steady expression intransfected human bladder carcinoma cell line T24 and its role in molecular regulatory mechanisms of telomerase, and to provide a new target gene for bladder cancer. Methods: PCR amplification was performed by using primers basedon the known gene sequence of hTERT. PCR productionwas cloned into plasmid pGEMT-T easy and the sequenceof mutant hTERT gene was analyzed. A recombinantmutant hTERT vector (pEGFP-hTERT) was constructed at the EcoR I and Sal I sites of the pEGFP-C1 vector. Aftertransfecting the fusion gene into bladder carcinoma cell line T24 by calcium phosphate-DNA coprecipitation, the steady expression of GFP-hTERT fusion protein was tested by fluorescent light microscopy. The proliferation changes ofbladder carcinoma cell line T24 were detected by lightmicroscopy and senescence correlated b-galactosidase staining. Results: Identification of pEGFP-hTERT byenzyme digestion showed that mutant hTERT fragment had been cloned into EcoR I and Sal I sites of the pEGFP-C1 vector. The steady expression of GFP-hTERT fusion protein was localized in the nucleus of transfected cells. Expression of senescence-associated b-galactosidase in transfected cells gradually increased with extended cultured time and cellgrowth was suppressed. Conclusion: The mutant-type hTERT gene suppresses the proliferation of bladder carcinoma cell line T24 by competitive effect on telomerase activity. This suggests that hTERT gene might be a suitable gene target for bladder cancer therapy.

Epidemiological Profile and Therapeutics Results of Bladder Carcinoma in Women

Introduction: The incidence of bladder cancer in women has steadily increased in recent years. The objective of our work is to study the epidemiological profile of bladder cancer in women in our population and the factors favoring its occurrence. Methods: This is a retrospective study carried out at the Mohamed Center VI for cancer treatment, involving 27 women during a two-year period from January 2019 to December 2020. We studied the epidemiological characteristics in these patients, the risk factors and the revealing symptoms. Results: The average age of patients was 67.5 years old and 66.66% of the patients were over 65 years old. One patient had in her antecedents a tumor of the upper excretory tract and 2 patients had an associated renal tumor. The notion of smoking was found in 3 patients. The time to treatment was less than 6 months in 44.4% of cases, between 6 and 12 months in 18.5% of cases, and more than a year in 37% of cases. The symptoms were marked by hematuria in 96.3% of cases, and irritative disorders in 81.4% of cases. In our series;the tumor was unifocal in 77.77% of the cases, and multifocal in the remainder of the cases. Urethrohydronephrosis was found in 74% of cases. Histologically, it was an urothelial carcinoma in 81.48% of cases. The tumor was locally advanced in 11.11% of cases, and metastatic in 11.11% of cases. Lymph node involvement was found in 33.33% of cases. 48.1% of cases underwent surgery. The operative procedure consisted of a partial cystectomy in 46.1% of cases, and only 18.5% was suitable for trimodal treatment. Conclusion: Few studies have addressed the epidemiology of bladder cancer in women due to the low incidence, but it is currently increasing steadily in recent years. These modifications are explained by the change in social habits in women.

The clinical significance of detection of specific CK-20 mRNA in peripheral venous blood from patients with bladder carcinoma

Objective To determine the diagnostic significance of detecting the specific epithelial keratin CK-20 mRNA in peripheral venous blood from patients with bladder carcinomas. Methods Reverse transcription coupled with two-step polymerase chain reaction (nested RT-PCR) was used to detect CK-20 mRNA expression in the peripheral blood from patients with blodder carcinomas. Results Detection of CK-20 mRNA expression was positive in 37 of 91 patients with bladder carcinoma (41 % ). Among 20 patients with distant metastasis, 17 were positive (85 % ). CK-20 mRNA was not detectable in the blood samples from 25 normal individuals. The frequency of positive CK-20 mRNA expression was signficantly higher in those with distant metastasis. Conclusion The presence of CK-20 mRNA expression in peripheral blood may be used as an early indicator of hematogenous metastasis of bladder carcinoma cells. 6 refs,1 tab.

Two Grams BID Is an Oral Dosage of Vitamin C to Reduce the Risk of Recurrence of Superficial Bladder Carcinoma

Background: Continuous exposure to millimolar (mM) Vitamin C (AA) in vitro kills cancer cells. For superficial bladder carcinoma (SBC), standard chemotherapy is instillation of Bacillus Calmette-Guerin. The recurrence rate with this therapy is 91%. But high dosage vitamins including AA reduced the recurrence to 41%. Aim: To determine the oral dosage of AA that causes the highest concentration of AA [AA] in the bladder. Method: We conducted a clinical trial of 14 people who took various dosages of AA, and analyzed the [AA] in their urine. Results: AA above 2 g twice a day was not absorbed. But that intake produced a bladder [AA] above 1 mM in all participants. Conclusion: Taking 2 g of AA BID will increase [AA] in the bladder to a level likely to kill cancer cells that cause SBC. Taking that dosage 2 consecutive days a week is likely to reduce the recurrence rate of SBC substantially.

Three cancers in the renal pelvis,bladder,and colon:A case report

BACKGROUND Multiple primary cancers are rare occurrences that can involve either metachronous or synchronous development.It is particularly rare for an individual to have more than two primary cancers.In this report,we present a case study of an elderly man who was diagnosed with three heterochronous cancers in the renal pelvis,bladder,and colon.CASE SUMMARY On December 30,2014,a 51-year-old Chinese man was admitted to our hospital with complaints of intermittent painless gross hematuria for the preceding week.A computed tomography(CT)scan revealed wall thickening in the left ureter’s upper segment,while a CT urography revealed a left renal pelvis tumor.A successful laparoscopic radical resection of the left renal pelvis tumor was subsequently performed at Shanghai Zhongshan Hospital in January 2015.The pathological findings after the surgery revealed a low-grade papillary urothelial carcinoma of the renal pelvis.The final pathological tumor stage was pT1N0M0.After surgery,this patient received 6 cycles of intravenous chemotherapy with gemcitabine and carboplatin,as well as bladder infusion therapy with gemcitabine.On December 18,2017,the patient was admitted once again to our hospital with a one-day history of painless gross hematuria.A CT scan showed the presence of a space-occupying lesion on the posterior wall of bladder.Cystoscopic examination revealed multiple tumors in the bladder and right cutaneous ureterostomy was performed under general anesthesia on December 29,2017.The postoperative pathological findings disclosed multifocal papillary urothelial carcinoma of the bladder(maximum size 3.7 cm×2.6 cm).The bladder cancer was considered a metastasis of the renal pelvis cancer after surgery.The pathological tumor stage was pT1N0M1.The patient refused chemotherapy after surgery.After another six years,the patient returned on February 28,2023,complaining of periumbilical pain that had lasted six days.This time,a CT scan of the abdomen showed a tumor in the ascending colon,but a subsequent colonoscopy examination indicated a tumor in the descending colon.On March 12,2023,a subtotal colectomy and an ileosigmoidal anastomosis were carried out under general anesthesia.Postoperative pathological findings revealed that all three tumors were adenocarcinomas.The final pathological tumor stage was pT3N0M0.The patient had an uneventful postoperative recovery and was discharged without complications.CONCLUSION The case of this elderly man presents a rare occurrence of metachronous primary cancers in the renal pelvis and colon.Bladder cancer is considered a metastasis of renal pelvis cancer after surgery.Optimal treatment can be implemented by evaluating the patient’s histological features,clinical history,and tumor distribution correctly.

Metachronous urothelial carcinoma in the renal pelvis,bladder,and urethra:A case report

BACKGROUND Urothelial carcinoma(UC)is a common malignancy of the urinary system that can occur anywhere from the renal pelvis to the proximal urethra.Most UCs are in the bladder and have multifocal growth.Upper urinary tract UC(UTUC),which occurs in the renal pelvis or ureter,accounts for only 5%to 10%of UCs.CASE SUMMARY In March 2015,a 70-year-old male who initially presented to a local hospital with a complaint of painless hematuria was diagnosed with UTUC of the right renal pelvis.The doctors administered radical nephroureterectomy and bladder cuff excision.Although the doctors recommended intravesical chemotherapy and regular follow-up,he rejected this advice.In December 2016,the patient presented at our hospital with dysuria.We identified UC in the residual bladder and administered radical cystectomy and left cutaneous ureterostomy.In November 2021,he presented again with urethral bleeding.We detected urethral UC as the cause of urethral orifice bleeding and administered radical urethrectomy.Since then,he has visited regularly for 6-mo follow-ups,and was in stable condition as of December 2022.CONCLUSION UTUC is prone to seeding and recurrence.Adjuvant instillation therapy and intense surveillance are crucial for these patients.

A Case of Papillary Thyroid Carcinoma Arising from Struma Ovarii and Extending into the Bladder

Ovarian goiter is a form of single tissue teratoma of the ovary, accounting for 2% - 3% of mature ovarian teratomas. Malignant transformation may occur in rare cases. Papillary thyroid-type carcinoma represents the most common type of malignant struma ovarii, followed by follicular carcinoma. Malignant struma ovarii is commonly seen in women in the fifth decade. The diagnosis is often made post-operatively after histological examination. Histology also helps assess tumor aggressiveness (mitoses, necrosis, poorly differentiated subtype, etc.). Given the rarity of these lesions, no therapeutic consensus or prognostic value had yet been formally established. We report herein, the case of a 76-year-old woman with a cystic tumor of the right ovary and a nodular lesion of the bladder. The clinical symptomatology is nonspecific, associating abdomino-pelvic pain and a right latero-uterine mass on abdominal palpation. After total hysterectomy with bilateral adnexectomy, the diagnosis of papillary carcinoma arising from struma ovarii and extending into the bladder was made. Through this observation, we suggest to discuss the anatomoclinical particularities of this rare pathological entity.

Results of Trans-Urethral Resection of Bladder (TURB) for the Treatment of Non-Infiltrating Bladder Tumors (NMIBT) in Musk in Bouake

Background: bladder tumors rank 2nd among urological tumors in sub-Saharan Africa, particularly in Côte d’Ivoire. Objective: to report the results of transurethral resection of the bladder (TURB) for the treatment of non-muscle-infiltrating bladder tumors (NMIBT) in Bouaké. Patients and methods: we conducted a cross-sectional, descriptive study of transurethral resection of the bladder (TURB) for the treatment of non-muscle-infiltrating bladder tumors (NMIBT) between January 2022 and April 2023. All patients and their families were informed in advance and had signed an informed consent form. All patients with a non-muscle-invasive bladder tumor confirmed by an initial TURB were included, and patients with a bladder tumor infiltrating the bladder muscle were excluded. Diagnosis was based on cystoscopy and anatomopathological examination of resection shavings. Parameters studied were: age, sex, risk factors, reason for consultation, clinical signs, cystoscopy findings, stage, grade, Evolution. Results: 17 patients with a mean age of 53.7 years (22-73 years) underwent trans-ureteral bladder resection to treat a non-muscle-infiltrating bladder tumor (NMIBT). Male gender predominated with 88.23% (n = 15), the majority of patients came from the ME region with 47.05% (n = 8), farmers were the most numerous (52.94%;n = 9). The most frequent reason for consultation was macroscopic hematuria with 64.1% (n = 11), risk factors were dominated by urinary bilharziasis with 70.58% (n = 12), physical examination was normal in 47.05% (n = 8). Hemoglobin (HB) levels were between 7.5 and 8.5 g/dl in 52.94% (n = 9). On cystoscopy, the tumor was budding in 76.45% (n = 13), the localization was trigonal in 52.9% (n = 9) and the base of implantation was sessile in 70.52% (n = 12). On ultrasound, the tumor was 3 cm or larger in 70.52% (n = 12). Therapeutically, 82.35% (n = 14) of patients received a blood transfusion. TURB was complete in the majority of cases 88.23% (n = 15). Squamous cell carcinoma was the most frequent histological type with 76.47% (n = 13). PTa and PT1 grade accounted for 23.52% (n = 4) and 76.47% (n = 13) respectively. High-grade PT1 accounted for 64.70% (n = 11). Follow-up to TURB was straightforward in 94.11% (n = 16). At three months post-TURB, seven patients presented a tumor recurrence, with 41.17% (n = 7) requiring a second TURB. At 6 months, follow-up noted 14 patients free of any clinical or endoscopic signs of bladder tumors. Conclusion: TURB is a safe and effective means of treating non-muscle-invasive bladder tumors.

Expression of KAI1/CD82 and MRP-1/CD9 in Transitional Cell Carcinoma of Bladder

The expression of KAI1/CD82 and MRP-1/CD9 in transitional cell carcinoma of bladder （TCCB） and its clinical significance were investigated. Immunohistochemistry was used to detect KAI1/CD82 and MRP-1/CD9 protein expression in 52 TCCB specimens. Correlation between the expression of KAI1/CD82 and MRP-1/CD9 to clinicopathologic factors was statistically analyzed. The results showed that the positive rate of KAI1/CD82 and MRP-1/CD9 in TCCB was 50% and 61.5%, respectively. The MRP-1/CD9 and KAI1/CD82 expression was significantly associated with grade of TCCB （P〈0.05）, but no correlation was found between MRP-1/CD9 or KAI1/CD82 expression and clinical stage of TCCB （P〉0.05）. The expression level of MRP-1/CD9 and KAI1/CD82 in recurrent TCCB samples was lower than that in non-recurrent samples （P〈0.05）. Meanwhile, the correlation between the KAI1/CD82 expression and MRP-1/CD9 expression was statistically significant （r=0.316, P〈0.05）. It was concluded that KAI1/CD82 and MRP-1/CD9 expression may be important prognostic indicators and potentially useful for assessing the biological behavior of TCCB.

ImmunoCytTM and cytology for diagnosis of bladder carcinoma：a meta analysis

Background Currently,cystoscopy and urine cytology are standard modalities in therapy monitoring and follow-up of bladder carcinoma （BC）.Cystoscopy is an invasive and uncomfortable procedure while cytology has a limited value because it is operator-dependent and has low sensitivity.This study was to assess the accuracy of ImmunoCyt in detecting BC by comparing it with cytology using systemic analyses of studies published in both English and Chinese.Methods Cochrane systematic evaluation was used to search through MEDLINE,EMBASE,Cochrane Library,CMCC,and CNKI for studies regarding ImmunoCyt and cytology for detection of BC.Data were extracted and analyzed by the software MetaDiSc 1.4.Results In total 42 relevant studies were searched,of which 15 were enrolled and 12 491 patients were included.Heterogeneity,except for threshold effects,was found within these studies.A meta-analysis was performed using the random effect model.Pooled accuracy indicators like sensitivity,specificity,and diagnostic odds ratio of ImmunoCytTM and cytology were 0.75 （0.73-0.77） vs.0.45 （0.43-0.48）,0.73 （0.72-0.74） vs.0.97 （0.96-0.97）,and 10.97 （7.53-15.99） vs.16.40 （10.57-25.46）,respectively.The sensitivity of both was increased with the increase of tumor grade and stage.The area under summary receiver operating characteristics curve was 0.834 4 and 0.853 4 and the Q index 0.766 7 and 0.785 3 for ImmunoCyt and cytology,respectively.Combination of both can obviously improve the accuracy of diagnosis.Conclusions ImmunoCyt has a high sensitivity in detecting BC,but its specificity is low.As an important adjunct,ImmunoCytTM can not replace cytology,but combined with cytology it could improve sensitivity with high specificity in the detection and postoperative monitoring of BC.

Expression of mi RNA-630 in Bladder Urothelial Carcinoma and Its Clinical Significance

Many studies informed that micro RNAs（mi RNAs） could function as diagnostic and prognostic indicators in several cancers. The aims of this study were to explore the expression of mi R-630 in bladder urothelial carcinoma and its clinical significance for the evaluation of cancer prognosis. A total of 116 patients with bladder urothelial carcinoma were obtained in this retrospective study between May, 2012 and Sep. 2015. Quantitative real-time PCR（q RT-PCR） was conducted to evaluate the expression level of mi R-630. The chi-square test was used to examine the associations between mi R-630 expression and the clinicopathological features. The Kaplan-Meier method was conducted to explore the survival status of urothelial carcinoma patients. The log-rank test was used to analyze differences in survival rate. The results showed an obvious increase in mi R-630 expression from normal bladder to bladder urothelial carcinoma（P=0.027）. Additionally, patients with higher mi R-630 expression had significantly shorter disease-free survival（DFS）（P=0.043） and overall survival（OS）（P=0.038） than those with lower mi R-630 expression. Furthermore, multivariate analysis revealed that up-regulation of mi R-630 was an independent prognostic factor for both DFS（P=0.042） and OS（P=0.046）. It was demonstrated that mi R-630 may be a novel and valuable prognostic factor for bladder urothelial carcinoma.

Multinucleated giant cells of bladder mucosa are modified telocytes:Diagnostic and immunohistochemistry algorithm and relation to PDL1 expression score

BACKGROUND Multinucleated giant cells(MGCs)in bladder carcinomas are poorly studied.AIM To describe the function,morphogenesis,and origin of mononuclear and MGCs in urothelial carcinoma(UC)of the bladder in Bulgarian and French patients.METHODS Urothelial bladder carcinomas(n=104)from 2016-2020 were analyzed retrospectively using immunohistochemical(IHC)and histochemical stain examination.Giant cells in the bladder stroma were found in 35.6%of cases,more often in highgrades.RESULTS We confirm that MGCs in the mucosa in UC of the bladder were positive for both mesenchymal and myofibroblast markers(vimentin,smooth muscle actin,Desmin,and CD34)and the macrophage marker CD68.Furthermore,IHC studies revealed the following profile of these cells:Positive for p16;negative for epithelial(CK AE1/AE3 and GATA-3),vascular(CD31),neural(PS100 and CKIT),cambial,blastic(CD34-blasts and C-KIT),and immune markers(IG G,immunoglobulin G4,and PD-L1);no proliferative activity,possess no specific immune function,and cannot be used to calculate the Combined Positive Score scale.CONCLUSION In conclusion,the giant stromal cells in non-tumor and tumor bladder can be used as a characteristic and relatively constant,although nonspecific,histological marker for chronic bladder damage,reflecting the chronic irritation or inflammation.Likewise,according to the morphological and IHC of the mono-and multinucleated giant cells in the bladder,they are most likely represent telocytes capable of adapting their morphology to the pathology of the organ.

The role of aberrant promoter hypermethylation of DACT1 in bladder urothelial carcinoma

The purpose of this study was to determine the relationship between hypermethylation of DACT1 gene pro-moter and lower mRNA expression in bladder urothelial carcinoma tissue.The methylation status of 29 urothelial carcinoma samples and 29 normal tissue samples were examined by methylation-specific polymerase chain reac-tion（MSP）.The DACT1 mRNA transcript levels and DACT1 protein levels in all samples were then evaluated to define the relationship between the methylation status of the DACT1 promoter and its expression at the transcrip-tional and translational levels.Decreased expression of DACT1 was detected in 89.66% of urothelial carcinomas（26/29;P 〈 0.005）.Promoter hypermethylation was found in 58.62%（17/29） urothelial carcinomas and 25%（7/29） normal tissues,respectively（P 〈 0.05）.DACT1 expression was lower in tissues where the DACT1 gene promoter was hypermethylated than in unmethylated tissues（0.25±0.17 vs 0.69±0.30,P 〈 0.05）.DACT1 gene hyper-methylation was closely related to tumor size,grade and stage（P 〈 0.05）.Our results indicate that silencing and downregulation of DACT1 mRNA may be implicated in carcinogenesis and the progression of bladder urothelial carcinoma,and may be a potential prognostic factor.

Infiltration Related miRNAs in Bladder Urothelial Carcinoma

This study aimed to investigate infiltration related microRNAs(miRNAs) in bladder urothelial carcinoma(BUC).Twenty patients with BUC were enrolled and divided into 2 groups according to infiltration or not:infiltrating BUC group(n=12) and non-infiltrating BUC group(n=8).Gene chip was used to detect infiltration related miRNAs in the BUC samples.In other recruited 17 patients with BUC who were divided into infiltrating BUC samples(n=14) and non-infiltrating BUC samples(n=3),and in 4 BUC cell lines(EJ,5637,T24 and BIU-87),the expression of miRNAs was assayed by using reverse transcription-polymerase chain reaction(RT-PCR).In infiltrating BUC group,as compared with non-infiltrating BUC group,there were 7 differentially expressed miRNAs:hsa-miR-29c,hsa-miR-200a,hsa-miR-378,hsa-miR-429,hsa-miR-200c and hsa-miR-141 were up-regulated,while hsa-miR-451 was down-regulated.In the BUC samples,the results of RT-PCR were consistent with those by the miRNA array.In the cancer cell lines,RT-PCR in T24 only revealed the similar expression pattern of miRNAs to that by the miRNA array.It is suggested that infiltration of BUC is related with different expression of miRNAs,which may provide a novel platform for further study on function and action mechanism of miRNAs.

MTHFR C677T polymorphisms are associated with aberrant methylation of the IGF-2 gene in transitional cell carcinoma of the bladder

The purpose of this study was to determine the relationship between methylation status of the insulin-like growth factor 2 （IGF-2） gene and methylenetetrahydrofolate reductase （MTHFR） C677T gene polymorphisms in bladder transitional cell carcinoma tissues in a Chinese population. The polymorphisms of the folate metabolism enzyme gene MTHFR were studied by restrictive fragment length polymorphism （RFLP）, PCR-based methods of DNA methylation analysis were used to detect the CpG island methylation status of the IGF-2 gene. The association between the methylation status of the IGF-2 gene and clinical characteristics, as well as MTHFR C677T polymorphisms, was analyzed. Aberrant hypomethylation of the IGF-2 gene was found in 68.3% bladder cancer tissues and 12.4% normal bladder tissues, respectively, while hypomethylation was not detected in almost all normal bladder tissues. The hypomethylation rate of the IGF-2 gene in cancer tissues was significantly higher in patients with lymph node metastasis than in those without lymph node metastasis （46.3% vs 17.2%, P = 0.018）. No association was found between aberrant DNA methylation and selected factors including sex, age, tobacco smoking, alcohol consumption and green tea consumption. After adjusting for potential confounding variables the variant allele of MTHFR C677T was found to be associated with hypomethylation of the IGF-2 gene. Compared with wildtype CC, the odds ratio was 4.33 （95% CI=1.06-10.59） for CT and 4,95 （95% CI=1.18-12.74） for TT. MTHFR 677 CC and CT genotypes might be one of the reasons that cause abnormal hypomethylation of the IGF-2 gene, and the aberrant CpG island hypomethylation of the IGF-2 gene may contribute to the genesis and progression of bladder transitional cell carcinoma.

Application of fluorescence in situ hybridization in the detection of bladder transitional-cell carcinoma: A multi-center clinical study based on Chinese population

Objective:To evaluate the diagnostic value of fluorescence in situ hybridization(FISH)in bladder cancer.Methods:We enrolled healthy volunteers and patients who were clinically suspected to have bladder cancer and conducted FISH tests and cytology examinations from August 2007 to December 2008.Receiver operating characteristic(ROC)curve analysis was performed and the area under curve(AUC)values were calculated for both the FISH and urine cytology tests.Results:A cohort of 988 healthy volunteers was enrolled to establish a reference range for the normal population.A total of 4807 patients with hematuria were prospectively,randomly enrolled for the simultaneous analysis of urine cytology,FISH testing,and a final diagnosis as determined by the pathologic findings of a biopsy or a surgically-excised specimen.Overall,the sensitivity of FISH in detecting transitional-cell carcinoma was 82.7%,while that of cytology was 33.4%(p<0.001).The sensitivity values of FISH for non-muscle invasive and muscle invasive bladder transitional-cell carcinoma were 81.7%and 89.6%,respectively(p=0.004).The sensitivity values of FISH for low and high grade bladder cancer were 82.6%and 90.1%,respectively(p=0.002).Conclusion:FISH is significantly more sensitive than voided urine cytology for detecting bladder cancer in patients evaluated for gross hematuria at all cancer grades and stages.Higher sensitivity using FISH was obtained in high grade and muscle invasive tumors.