Low-dose abiraterone plus Olaparib as a late-line treatment for mCRPC patients without BRCA1/2 mutations: a multicenter retrospective pilot study.

Although overall survival data are still premature, the PROpel study found radiological progression-free survival (PFS) benefits of abiraterone and olaparib in patients with metastatic castration-resistant prostate cancer (mCRPC). However, for patients who have not been genetically tested or lack BRCA1/2 mutations (BRCAm), this combination therapy has been questioned as a first-line conventional treatment for mCRPC, mainly due to significant health economics and side effects. In our retrospective study, we found that treatment with low-dose abiraterone plus olaparib as a late-line treatment for mCRPC could lead to prostate-specific antigen (PSA) and symptom PFS in selective cases even without BRCAm. The median PSA-PFS was 8 months (IQR: 6.5-11.5), with a median follow-up duration of 39.0 months (IQR: 27.5-64.5). Gene tests were conducted in all patients, identifying non-BRCA mutations through ctDNA testing (24%), tumor tissue testing (12%), or both (64%). Adverse events occurred in 72% of patients, with 16% experiencing Grade ≥ 3 events. Common adverse events included anemia (64%), decreased appetite (48%), and fatigue (25%). Our findings support low-dose abiraterone plus olaparib as a potential option for mCRPC patients without BRCAm, offering manageable safety and efficacy profiles.

Androgen deprivation therapy plus apalutamide as neoadjuvant therapy prior radical prostatectomy for patients with unresectable prostate cancer.

Whether neoadjuvant therapy confers a survival benefit in advanced prostate cancer (PCa) remains uncertain. The primary endpoints of previous retrospective and phase II clinical studies that used neoadjuvant therapy, including androgen deprivation therapy combined with new-generation androgen receptor signaling inhibitors or chemotherapy, were pathological downstaging, progression-free survival, prostate-specific antigen relief, and local symptom improvement. To the best of our knowledge, no studies have explored the efficacy and safety of neoadjuvant therapy in improving the surgical resection rate in cases of unresectable primary tumors of PCa. We first designed this retrospective study to evaluate the potential value of apalutamide as neoadjuvant therapy in improving the resectability rate of radical prostatectomy (RP). We initially reported 7 patients with unresectable primary lesions who underwent neoadjuvant apalutamide treatment for a median of 4 months, and all of them successfully underwent RP treatment. Our study supported apalutamide as neoadjuvant therapy, which helped improve RP's success rate and did not significantly increase perioperative complications, and the neoadjuvant therapy was controllable. Our findings' clinical value and benefit for survival still need further clinical research to confirm.

Pharmarcomechanical thrombectomy combined with transluminal balloon angioplasty for treating transplant renal vein thrombosis.

Renal vein thrombosis (RVT) is a rare vascular complication that occurs after renal transplantation and usually results in irreversible kidney damage and graft loss. We report the case of a patient who underwent right iliac fossa allogeneic kidney transplantation and developed RVT combined with ipsilateral thrombosis from the popliteal to the femoral veins, with extension to the common iliac veins, 4 months after transplantation. Under unfractionated heparin anticoagulation, an Aegisy (Life Tech Scientific Co., Ltd., Shenzhen, China) vena cava filter was placed to prevent pulmonary embolism. Percutaneous mechanical thrombectomy combined with balloon angioplasty was performed to aspirate the thrombus and successfully dilate the narrow venous lumen. The patient's renal function was restored postoperatively. Ultrasonography showed the allograft and ipsilateral lower extremity deep veins to be fluent and patent. To conclude, in patients with RVT after renal transplantation, percutaneous mechanical thrombectomy in conjunction with balloon angioplasty can be performed with desirable outcomes and no severe adverse effects. This method reduces the risk of bleeding from exposure to systemic intravenous thrombolysis and avoids surgery-associated trauma.

Presence of lymph nodes and metastasis within prostatic anterior fat pad in radical prostatectomy patients: A single Chinese institution experience and literature review.

INTRODUCTION: Limited data from China, aim to investigate the incidence and the risk fctors of lymph node metastases in the prostatic anterior fat pad (PAFP). MATERIAL AND METHODS: Patients underwent radical prostatectomy (RP) were enrolled between March 2020 to December 2022 at a single institution. Separate pathological analysis of PAFP was performed within this area. Univariate analysis and Multivariate analysis were performed to determine the risk factor of PAFP metastasis. RESULT: A total of 255 patients were included. The study revealed an average age of 67.72 ± 7.07 years, with a mean total tumor volume of 41.54 ± 23.79 mL, and an average Pre-op PSA of 16.85 ng/mL. Clinical T stage was divided into T2, T3, and T4 (226, 25, 4 cases, respectively), while the Clinical M stage was categorized as M0 and M1 (248 and 7 cases, respectively). Out of the patients with PAFP, 19 (7.45 %) had lymph node in PAFP, and 3 (1.18 %) patients had metastases. In the univariate and multivariate analysis, Clinical M stage and anterior primary tumor were found to be a significant high-risk factor. Among the other 15 studies, six examined the risk factors associated with it, including anterior tumors, higher tumour volume, intermediate or high risk prostate cancer. CONCLUSION: Due to the low proportion of lymph node involvement (7.45 %) and rare tumor metastasis (1.18 %), routine separate pathological analysis of PAFP is not recommended in all RP patients unless there are anterior tumors, higher tumor volume, or intermediate/high risk prostate cancer.

Comparison of Survival Outcomes and Risk Factors Between Ductal Carcinoma of the Prostate and Acinar Adenocarcinoma of the Prostate: A Population-based Propensity Score-matching Study.

Background: Ductal carcinoma of the prostate (DCP) is a rare type of prostate cancer (PCa) with a higher degree of infiltration and worse prognosis than acinar adenocarcinoma of the prostate (ACP). Previous reports comparing DCP and ACP have not been very reliable and involved small sample sizes. Objective: To assess differences in mortality between ACP and DCP in a large-scale study. Design setting and participants: Data were downloaded from the Surveillance, Epidemiology, and End Results database in June 2022. Data for 823 939 patients diagnosed with PCa from 2004 to 2019 were examined, excluding cases with survival data missing or pathological types other than DCP and ACP. Outcome measurements and statistical analysis: Prognostic and risk factors for DCP were analyzed by generating a propensity score-matched cohort of DCP and ACP cases (1:5). Adjusted Cox models were constructed to determine hazard ratios (HRs) with 95% confidence intervals (CIs) for cancer-specific mortality (CSM) and overall mortality (OM). Results and limitations: A total of 822 607 cases (99.8%) has ACP and 1332 (0.2%) had DCP. In comparison to ACP, age at diagnosis was significantly lower for DCP (≤66 yr: 38.0% vs 50.7%; p < 0.001) and a higher proportion of DCP patients distant metastases (13.7% vs 5.1%; p < 0.001). In comparison to the ACP group, significantly higher proportions of the DCP group underwent surgery (66.1% vs 38.1%; p < 0.001), radiotherapy (13.7% vs 3.1%; p < 0.001), or systemic therapy (18.2% vs 3.3%; p < 0.001). However, the median overall survival time was significantly shorter for DCP patients (44.0 vs 73.0 mo; p < 0.001). DCP patients also had higher risk of CSM (HR 2.07, 95% CI 1.68-2.56; p < 0.001) and OM (HR 2.73 95% CI 2.42-3.08; p < 0.001) after propensity score matching to adjust for the influence of baseline variables. Subgroup analysis showed that DCP patients who had surgical treatment had better CSM than those without surgery, while DCP patients with regional and lower stage had better OM than those with distant stage (both p < 0.05 for interaction). Conclusions: The risk of CSM and OM is significantly higher for DCP than for ACP. Earlier detection (lower stage) and surgical treatment are beneficial factors for DCP prognosis. Patient summary: We studied survival rates for two different types of prostate cancer. We found that survival is worse for the rarer ductal carcinoma of the prostate (DCP) than for the more common acinar adenocarcinoma of the prostate. Both early diagnosis when the cancer is at a lower stage and surgical treatment are beneficial for survival in patients with DCP.

Blood circulating exosomes carrying microRNA-423-5p regulates cell progression in prostate cancer via targeting FRMD3.

This study explored the role of circulating exosomal microRNA-423-5p in the progression of PCa and its molecular mechanism. First, based on the microarray analysis, microRNA-423-5p was at a high expression level in PCa peripheral blood samples. It was demonstrated that microRNA-423-5p expression in serum exosomes of PCa patients was notably higher than that in healthy people as revealed by qRT-PCR. Further studies indicated that overexpressing microRNA-423-5p promoted cell progression of PCa. Microarray analysis and luciferase gene reporter assay illustrated that FRMD3 was targeted by microRNA-423-5p, and its expression was down-regulated by microRNA-423-5p. While FEMD3 knockdown would reverse the repressive effect of silencing microRNA-423-5p on PCa cell functions. In addition, it was exhibited that exosomes carrying microRNA-423-5p could internalize into PCa cells by labeling and tracing exosomes. Cell function assays and animal experiments manifested those exosomes carrying microRNA-423-5p could enhance PCa cell proliferation, migration, and invasion in vivo. In conclusion, this study indicated that blood circulating exosomal microRNA-423-5p played important roles in PCa cell functions, and illustrated the molecular mechanism of microRNA-423-5p as an oncogene in PCa.

Applicability of 14 Formulas for Estimating Glomerular Filtration Rate in the Evaluation of Renal Function before and after Nephron-Sparing Surgery in Patients with Renal Tumors.

To compare the applicability of 14 equations of estimating glomerular filtration rate (eGFR) before and after nephron-sparing surgery (NSS) for renal function assessment of patients with renal tumors. Preoperative and postoperative GFR is measured by emission computed tomography (ECT) with 99mTc-DTPA as an imaging agent as reference GFR (rGFR) to compare with all formulas. Spearman correlation analysis and Bland-Altman agreement analysis were used to evaluate the correlation between rGFR and eGFR1 to 14 before and after surgery. A total of 50 cases including 22 males and 28 females were included. The results of preoperative eGFR1-14 correlated with rGFR (P < 0.05). The calculation results of all estimation formulas have a significant correlation with preoperative GFR. Preoperative MDRD-I, CKD-EPI SCysC, and FAS Scr-SCysC have good consistency. The CG formula has the highest precision and FAS Scr-SCysC has the highest accuracy. A total of 30 patients followed up after surgery, and postoperative rGFR correlated with CG, CKD-EPI, FAS, and BIS formulas (P < 0.05). But postoperative rGFR has no significant correlation with MDRD and Schwartz (P > 0.05). Postoperative CKD-EPI Scr-SCysC has best consistency, and FAS Scr-SCysC has the highest accuracy and precision. Our data suggest that eGFR equations evaluated by both serum creatinine (Scr) and cystatin C (SCysC) is not necessarily better than those evaluated by one of them alone. Among all enrolled equations, FAS Scr-SCysC is the best one to evaluate postoperative GFR in patients with renal tumors.

The Application of Scrotoscope-Assisted Minimally Invasive Excision for Epididymal Mass: An Initial Report.

Background: To compare the middle-term efficacy and safety results between scrotoscope-assisted (SA) minimally invasive excision and traditional open excision (OE) for the treatment of epididymal mass. Methods: A total of 253 males with surgery excision of epididymal mass from 2012 to 2018 were included in this retrospective study. Patients were divided into two groups: the traditional OE group and the SA group. Patient demographics and intraoperative and postoperative outcomes were obtained and compared between these two groups. Results: About 174 patients (68.8%) underwent SA, and the other 79 (31.2%) underwent OE. Demographic data were similar between the two groups. Compared with OE surgery, SA could significantly shorten the operating time (19.4 ± 4.1 vs. 53.8 ± 12.9 min), reduce blood loss (5.3 ± 1.5 vs. 21.3 ± 5.6 ml), and downsize the operative incision (1.5 ± 0.3 vs. 4.5 ± 0.8 cm). Additionally, postoperative complications were significantly less occurred in the SA group than those in OE (15.5% vs. 21.5%), in particular scrotal hematoma (1.7% vs. 12.7%) and incision discomfort (2.8% vs. 6.3%). Patients in the SA group had a significantly higher overall satisfaction score (94.8 ± 3.7 vs. 91.7 ± 4.9) and a significantly shorter length of hospital stay (4.1 ± 0.9 vs. 5.0 ± 1.5 days) than those in the OE group. No postoperative testicular atrophy occurred in the SA group. Conclusion: SA is emerging as a novel and effective option with promising perspectives for epididymal mass therapy.

Increasing the efficiency and precision of prime editing with guide RNA pairs.

The recently reported prime editor (PE) can produce all types of base substitution, insertion and deletion, greatly expanding the scope of genome editing. However, improving the editing efficiency and precision of PE represents a major challenge. Here, we report an approach termed the homologous 3' extension mediated prime editor (HOPE). HOPE uses paired prime editing guide RNAs (pegRNAs) encoding the same edits in both sense and antisense DNA strands to achieve high editing efficiency in human embryonic kidney 293T cells as well as mismatch repair-deficient human colorectal carcinoma 116 cells. In addition, we found that HOPE shows greatly improved product purity compared to the original PE3 system. We envision that this enhanced tool could broaden both fundamental research and therapeutic applications of prime editing.

Preliminary assessment of a portable Raman spectroscopy system for post-operative urinary stone analysis.

PURPOSE: We aimed to evaluate the reliability of a portable device that applies Raman spectroscopy at an excitation wavelength of 1064 nm for the post-operative analysis of urinary stone composition. MATERIALS AND METHODS: Urinary stone samples were obtained post-operatively from 300 patients. All samples were analyzed by the portable Raman spectroscopy system at an excitation wavelength of 1064 nm as well as by infrared spectroscopy (IR), and the results were compared. RESULTS: Both Raman spectroscopy and IR could detect multiple stone components, including calcium oxalate monohydrate, calcium oxalate dihydrate, calcium phosphate, uric acid, cystine, and magnesium ammonium phosphate hexahydrate. The results from 1064-nm Raman analysis matched those from IR analysis for 96.0% (288/300) of cases. Although IR detected multiple components within samples more often than Raman analysis (239 vs 131), the Raman analysis required less time to complete than IR data acquisition (5 min vs 30 min). CONCLUSIONS: These preliminary results indicate that 1064-nm Raman spectroscopy can be applied in a portable and automated analytical system for rapid detection of urinary stone composition in the post-operative clinical setting. TRIAL REGISTRATION: Chinese Clinical Trail Register ID: ChiCTR2000039810 (approved WHO primary register) http://www.chictr.org.cn/showproj.aspx?proj=63662 .

Mechanism of tumor­derived extracellular vesicles in regulating renal cell carcinoma progression by the delivery of MALAT1.

Renal cell carcinoma (RCC) is a major healthcare burden globally. Tumor­derived extracellular vesicles (EVs) contribute to the formation of a pro­metastatic microenvironment. In the present study, we explored the role and mechanism of RCC cell 786­O­derived EVs (786­O­EVs) in RCC. First, 786­O­EVs were extracted and identified, and EV internalization of RCC cells was observed. RCC cell malignant behaviors and long noncoding RNA (lncRNA) metastasis­associated lung adenocarcinoma transcript 1 (MALAT1) expression patterns were detected before and after 786­O­EV treatment. MALAT1 was intervened to evaluate RCC cell behaviors. The downstream mechanism involving MALAT1 was predicted. In addition, the relationship among MALAT1, transcription factor CP2 like 1 (TFCP2L1) and ETS proto­oncogene 1, transcription factor (ETS1) was analyzed. TFCP2L1 expression patterns were measured after 786­O­EV exposure. Tumor xenograft formation assay and lung metastasis model were adopted to verify the role of 786­O­EVs in vivo in RCC. It was found that 786­O­EVs could be internalized by RCC cells. 786­O­EVs promoted RCC cell malignant behaviors, accompanied by elevated MALAT1 expression levels. The 786­O­EVs with MALAT1 knockdown attenuated the promotive effect of sole 786­O­EVs on RCC cells. MALAT1 located ETS1 in the TFCP2L1 promoter and negatively regulated TFCP2L1, and ETS1 protein could specifically bind to MALAT1. 786­O­EVs enhanced the binding of ETS1 and the TFCP2L1 promoter and decreased TFCP2L1 expression. In vivo, 786­O­EVs promoted tumor growth and RCC lung metastasis, which was suppressed following inhibition of MALAT1. Our findings indicated that 786­O­EVs promoted RCC invasion and metastasis by transporting MALAT1 to promote the binding of transcription factor ETS1 and TFCP2L1 promoter.

The effect of the papillary renal cell carcinoma subtype on oncological outcomes.

The study aimed to compare the clinicopathological features and prognosis between type I and type II papillary renal cell carcinoma (PRCC) and to investigate whether the subtypes of PRCC would affect oncological outcomes. A total of 102 patients with PRCC were recruited, of which 42 were type I PRCC and 60 type II. The clinicopathological features and oncologic outcomes of the patients were evaluated. The type II cases had a higher WHO/ISUP grading (P < 0.001), T (P = 0.003), N (P = 0.010) stage and stage grouping (P = 0.011) than the type I. During a median follow-up period of 61.4 months, 1-year cancer specific survival (CSS) of the type I was 100%, 5-year CSS was 95.2%, the 1-year CSS of the type II was 96.2%, and 5-year CSS was 75.7%. The univariate analysis showed that subtype, symptoms, TNM, stage grouping, WHO/ISUP grading and surgical methods appeared to affect prognosis of the patients with PRCC. However, multivariate analysis revealed that only stage grouping was the independent risk factor. After the stage grouping factor was adjusted for the analysis, there were no statistically significant differences in CSS (P = 0.214) and PFS (P = 0.190) between the localized type I and type II PRCC groups. Compared with type I PRCC, type II had higher pathological T, N stage and WHO/ISUP grading. However, it was the Stage grouping that made a great difference to oncological outcomes, rather than the subtype of PRCC.

Modified three-level techniques of retroperitoneal laparoscopic procedures to treat adrenal lesions for patients with BMI ≥ 25 Kg/m2.

Objective: To evaluate the modified Zhang's 'three-level' technique of retroperitoneal laparoscopic adrenalectomy (RLA) to treat adrenal lesions for patients with BMI of 25-30 Kg/m2. Methods: A retrospective analysis was performed in all patients with BMI of 25-30 Kg/m2 in our hospital from January 2014 to December 2019. Those who underwent laparoscopic adrenal surgery were divided into two groups on the basis of the technique used: the Zhang's technique (the ZT group) and the modified technique (the MT group). Results: Herein, 170 operations were included (ZT, 91 patients; MT, 79 patients). RLA was successfully performed in all of them. Compared with the ZT group patients, the MT group patients showed shorter operation time (p = 0.007), lesser intraoperative blood loss (p = 0.023), shorter operation time, earlier postoperative diet recovery (p < 0.001), shorter postoperative drainage time (p < 0.001) and shorter postoperative hospitalization period (p = 0.001). It was also worth noting that the unplanned total adrenalectomy rate was significantly less in the MT group than in the ZT group (0% vs. 10.8%, p = 0.020). There was no significant difference in the complications between the two groups (3.3% vs. 2.5%, p = 0.567). Conclusions: We found that MT was a beneficial retroperitoneal laparoscopic treatment for adrenal lesions in patients who had a BMI of 25-30 Kg/m2. It may provide a reference for the treatment of adrenal surgical diseases in such patients.

Scrotoscopy and traditional open surgery shows a high degree of consistency in the diagnosis of testicular torsion: An initial report.

Traditional open surgery (OS) is usually necessary when testicular torsion (TT) cannot be excluded by scrotal ultrasound. Scrotoscopy has been used as a minimally invasive technique to diagnose or treat scrotal diseases, and it may also play a role in diagnosing TT.A retrospective analysis was performed for patients with TT to evaluate the consistency of scrotoscopy and OS in the diagnosis of TT. In the cases where preoperational Color Doppler ultrasonography was performed, scrotoscopy, open surgery, and confirmed TT were included for future analysis.A total of 43 patients were studied. Twisted testes were retained in 11 cases (25.59%), and the remaining 32 patients (74.41%) underwent orchiectomy. There were significant differences in the diagnostic value between the grading of scrotoscopy and ultrasound, as well as between ultrasound grading and blood supply grading (BSG) (both P < .05). However, no significant difference was observed between the grading of scrotoscopy and BSG in traditional OS (P > .05), but a high degree of consistency existed between scrotoscopy grading and BSG in traditional OS (Kappa = 0.733, P ≤ .001).Our limited data indicate that the diagnosis of testicular torsion by scrotoscopy is highly consistent with that of traditional surgical exploration. Therefore, further studies are necessary to confirm its application value in the future. Scrotoscopy may have potential application value for the patients whom testicular torsion are insufficiently diagnosed but cannot be excluded.

Dioscin Promotes Prostate Cancer Cell Apoptosis and Inhibits Cell Invasion by Increasing SHP1 Phosphorylation and Suppressing the Subsequent MAPK Signaling Pathway.

Dioscin possesses antioxidant effects and has anticancer ability in many solid tumors including prostate cancer (PCa). Nevertheless, its effect and mechanism of anti-PCa action remain unclear. The tyrosine protein phosphatase SHP1, which contains an oxidation-sensitive domain, has been confirmed as a target for multicancer treatment. Further studies are needed to determine whether dioscin inhibits PCa through SHP1. We performed in vitro studies using androgen-sensitive (LNCaP) and androgen-independent (LNCaP -C81) cells to investigate the anticancer effects and possible mechanisms of dioscin after administering interleukin-6 (IL-6) and dihydrotestosterone (DHT). Our results show that dioscin inhibited cell growth and invasion by increasing SHP1 phosphorylation [p-SHP1 (Y536)] and inhibiting the subsequent P38 mitogen-activated protein kinase signaling pathway. Further in vivo studies confirmed that dioscin promoted caspase-3 and Bad-related cell apoptosis in these two cell lines. Our research suggests that the anticancer effects of dioscin on PCa may occur through SHP1. Dioscin may be useful to treat androgen-sensitive and independent PCa in the future.

miR-223-5p targeting ERG inhibits prostate cancer cell proliferation and migration.

Ectopic expression of miR-223-5p, the lagging strand of miR-223 duplex, has been reported acting as anti-tumor miRNA in many cancers. How miR-223-5p influencing prostate cancer (PCa) remains obscure and worth of experimental investigation. In this study, the expressions of miR-223-5p and ERG in common PCa cell lines were detected and compared to RWPE-1, respectively. Then luciferase reporter assay was performed to verify whether miR-223-5p could specifically target and regulate ERG. Further discovery ERG's role in the PCa oncogenesis was also conducted by up or down regulating miR-223-3p expression. We found miR-223-5p was significantly down-regulated in DU145, while it was only up-regulated in LNCaP. Similarly, ERG expression remarkably decreased in both PC-3 and DU145 than that in RWPE-1, but significantly increasing in LNCaP. Luciferase assay demonstrated slightly decreased ERG expression after miR-223-5p-mimics but significantly increased ERG expression after miR-223-5p-inhibtor. Using gene interference, we further confirmed that both ERG mRNA and protein expressions were decreased in all PCa lines transfected ERG siRNA, but increasing in both DU145 and LNCaP cells with miR-223-5p antisense oligonucleotides. MTT assay, Transwell invasion and migration assay supported the function of ERG in PCa oncogenesis. We revealed tumor suppressive abilities of miR-223-5p in PCa by negatively targeting ERG gene. It could serve as a fundamental supplement and extension of our previous study about miR-223-3p in PCa, revealing the coordinative regulation between miR-223-5p and miR-223-3p in PCa cell biological behaviors. Exploration of miR-233-duplex orientated pathway networks may help us develop novel potential therapeutic options for PCa.

Circ_0039569 promotes renal cell carcinoma growth and metastasis by regulating miR-34a-5p/CCL22.

Circular RNAs (circRNAs) belong to non-coding RNAs and are known as key regulators in gene regulation. CircRNAs involve in the various biological processes of cancer. However, the functions of circRNAs in renal cell carcinoma (RCC) are still not clear. In this study, the circRNA expression profile was performed in the RCC tissues by microarray. There were 35 significantly expressed circRNAs with more than 5 folds from microarray analysis. Hsa_circ_0039569 (circ_0039569) was verified to be up-regulated in RCC and cells compared with the controls by real time RT-PCR. The assays of cellular functions showed that circ_0039569 down-regulation suppressed the proliferation and metastasis of RCC cells. The molecular mechanism of circ_0039569 in RCC cells showed that circ_0039569 promoted RCC progression by up-regulating CCL22 expression via down-regulating miR-34a-5p. Taken together, the study indicated that circ_0039569 played an important role in RCC cell survival and metastasis, which suggested that an oncogenic role of circ_0039569 in RCC progression.

Irradiated Bladder Cancer Cells Expressing both GM-CSF and IL-21 versus Either GM-CSF or IL-21 Alone as Tumor Vaccine in a Mouse Xenograft Model.

Previous studies have established the efficacy of irradiated cancer cells overexpressing GM-CSF or IL-21 as a vaccine. Here we examined whether the vaccine efficacy was greater when both factors were overexpressed together. MB49 bladder cancer cells were transfected with expression plasmid pT7TS encoding mouse GM-CSF and human IL-21, and then irradiated with 100 Gy at 4 days later. The cells (1×107 per animal) were injected subcutaneously into C57BL/6 mice at 0, 4, 8, and 12 days after inoculation with MB49 tumor xenografts. Control animals were injected with MB49 cells transfected with pT7TS encoding GM-CSF or IL-21 on its own. Tumor growth was monitored for 45 days and compared among the groups using repeated-measures ANOVA. Vaccination with irradiated MB49 cells did not affect xenograft growth. Vaccination with irradiated cells overexpressing GM-CSF or IL-21 alone significantly inhibited tumor growth and led to significantly more CD4+ CD8+ T cells and fewer CD4+ Foxp3+ T cells in the spleen and xenograft. These effects were even greater following vaccination with irradiated cells overexpressing both GM-CSF and IL-21. Irradiated bladder cancer cells overexpressing both GM-CSF and IL-21 are more effective than cells expressing either factor alone as a vaccine against bladder cancer.