Mir-135a enhances cellular proliferation through post-transcriptionally regulating PHLPP2 and FOXO1 in human bladder cancer.

BACKGROUND: Bladder cancer is the most common malignancy in urinary system and the ninth most common malignancy in the world. MicroRNAs (miRNAs) are small, non-coding RNAs that regulate gene expression by targeted repression of transcription and translation and play essential roles during cancer development. We investigated the expression of miR-135a in bladder cancer and explored its bio-function during bladder cancer progression. METHODS: The expression of miR-135a in bladder cancer cells and tissues are performed by using Real-time PCR assay. Cell viability assay (MTT assay), colony formation assay, anchorage-independent growth ability assay and Bromodeoxyuridine labeling and immunofluorescence (BrdUrd) assay are used to examine cell proliferative capacity and tumorigenicity. Flow cytometry analysis is used to determine cell cycle progression. The expressions of p21, p27, CyclinD1, Ki67, PHLPP2 and FOXO1 are measured by Western blotting assay. Luciferase assay is used to confirm whether FOXO1 is the direct target of miR-135a. RESULTS: miR-135a is upregulated in bladder cancer cells and tissues. Enforced expression of miR-135a promotes bladder cancer cells proliferation, whereas inhibition of miR-135a reverses the function. Furthermore, for the first time we demonstrated PHLPP2 and FOXO1 are direct targets of miR-135a and transcriptionally down-regulated by miR-135a. Suppression of PHLPP2 or FOXO1 by miR-135a, consisted with dysregulation of p21, p27, Cyclin D1 and Ki67, play important roles in bladder cancer progression. CONCLUSION: Our study demonstrates that miR-135a promotes cell proliferation in bladder cancer by targeting PHLPP2 and FOXO1, and is performed as an onco-miR.

Bladder preservation in complicated invasive urothelial carcinoma following treatment with cisplatin/gemcitabine plus tislelizumab: A case report.

BACKGROUND: Invasive urothelial carcinoma (UC) with squamous and glandular differentiation is a highly malignant and complicated pathological subtype, and the standard care is radical cystectomy (RC). However, urinary diversion after RC significantly reduces patient quality of life, thus bladder-sparing therapy has become a research hotspot in this field. Recently, five immune checkpoint inhibitors have been approved for systemic therapy of locally advanced or metastatic bladder cancer by the Food and Drug Administration, but the efficacy of immunotherapy combined with chemotherapy for invasive UC is still unknown, especially for pathological subtypes with squamous and glandular differentiation. CASE SUMMARY: We report the case of a 60-year-old male who complained of repetitive painless gross hematuria and was diagnosed with muscle-invasive bladder cancer with squamous and glandular differentiation, defined as cT3N1M0 according to the American Joint Committee on Cancer, who had a strong desire to preserve the bladder. Immunohistochemical staining revealed that programmed cell death-ligand 1 (PD-L1) expression in the tumor was positive. Thus, a transurethral resection to maximize removal of the bladder tumor was performed under cystoscopy, and the patient subsequently received a combination of chemotherapy (cisplatin/gemcitabine) and immunotherapy (tislelizumab) treatment. No tumor recurrence in the bladder was observed following pathological and imaging examination after 2 cycles and 4 cycles of treatment, respectively. The patient achieved bladder preservation and has been tumor-free for more than two years. CONCLUSION: This case shows that the combination of chemotherapy and immunotherapy might be an effective and safe treatment strategy for PD-L1 expression positive UC with divergent histologic differentiation.

[Clinical characteristics and surgical treatment for localized Castleman's disease].

OBJECTIVE: To investigate the clinical characteristics and treatment of localized Castleman's disease (CD), and review the literatures to improve the diagnosis and management of this disease. METHODS: The clinical symptoms, histopathology, CT, MRI findings and results of surgery in 20 patients with localized CD were evaluated retrospectively. RESULTS: The average age of the patients was 37.7 years. The lesions were located in the retroperitoneal space (9 cases), mediastinum (7 cases), pelvic cavity (1 case), neck (1 case), upper arm (1 case), and axillary (1 case). All patients underwent surgical resection, including 9 cases for retroperitoneal resection (6 cases had open operation and 3 cases laparoscopic resection) and 7 cases for mediastinal resection (open operation in 5 cases and thoracoscopic resection in 2 cases). The Castleman's disease was confirmed by histopathology. There were hyaline vascular type of CD in 17 cases, plasma cell type of CD in 1 case, and mixed cellularity type of CD in 2 cases. The duration of follow-up ranged from 12 to 165 months for 16 cases. Among them 15 patients were alive without recurrence, and 1 case had recurrence in the primary site at 47 months after the operation. CONCLUSIONS: Patients with Castleman's disease have no typical clinical symptoms and have normal laboratory results. The majority of patients are of hyaline vascular type of the disease. Imaging examination is helpful to diagnosis, and the final diagnosis depends on pathologic examination. Complete surgical resection of the tumor is the best treatment for localized Castleman's disease.

[Factors associated with increased blood loss in patients undergoing radical cystectomy in a contemporary series].

OBJECTIVE: To evaluate factors predictive of blood loss in radical cystectomy in a contemporary series. METHODS: From December 1996 to December 2008, clinical data of 233 patients who underwent radical cystectomy were reviewed retrospectively. Various preoperative and operative factors were assessed for their association with blood loss using univariate, multivariate regression and correlation analysis. RESULTS: One hundred eighty-one patients underwent open radical cystectomy and 52 cases were treated by laparoscopic radical cystectomy. Overall mean operative time was (339 ± 84) minutes, and mean blood loss was (818 ± 756) ml. On univariate analysis, body mass index (F = 9.039), history of pelvic operation (t = -4.365), anesthetic techniques (t = 3.125), surgical type (t = 6.643), use of Ligasure (t = 6.923), and urethra resection (t = -1.984) correlated with blood loss. However, multiple linear regression showed that body mass index (R(2) = 0.256, P < 0.001), history of pelvic operation (R(2) = 0.222, P < 0.001), use of Ligasure (R(2) = 0.172, P < 0.001), and surgical type (R(2) = 0.271, P = 0.027) were significant predictors of blood loss. The transfusion was required in 176 of 233 patients (75.5%) with a median requirement of (649 ± 569) ml. Likewise logistical regression analysis revealed that older age (OR = 3.2, P = 0.010), female gender (OR = 33.7, P = 0.013), anemia (OR = 6.6, P = 0.039), increased blood loss (OR = 14.3, P < 0.001), open radical cystectomy (OR = 6.4, P = 0.036) and nonuse of Ligasure (OR = 10.1, P < 0.001) were predictors of transfusion need. CONCLUSIONS: Increased body mass index, history of pelvic operation, open radical cystectomy, and non-use of Ligasure were independent predictors of increased blood loss during radical cystectomy. Such a prediction formula has an important role in identifying high risk patient for increased blood loss and transfusion need before radical cystectomy.

[The analysis of perioperative complications during radical cystectomy using a standardized reporting system].

OBJECTIVES: To analyze the perioperative complications of radical cystectomy using a standardized reporting methodology. METHODS: The clinical data of 233 cases of radical cystectomy from January 1996 to December 2008 were reviewed. Two hundred male patients and 33 female patients were included. The mean age was 58.9 years old. All complications within 30 days of surgery were recorded and classified using a 5-grade modification of the Clavien system. RESULTS: Overall mean operative time was 339 (170 - 610) minutes, and mean blood loss was 818 (range 100 to 3500) ml. Of the 233 subjects at least 1 postoperative complication developed in 84 (36.1%), including five cases of intraoperative complications. According to the modified Clavien system, 27 patients (11.6%) had grade 1, 38(16.3%) had grade 2, 16(6.9%) had grade 3, and 3(1.7%) had grade 4 complications. The most frequent complication was gastrointestinal complications (15.9%), then the incision-related complications (15.0%) and lung infections (4.7%). An association between hypoproteinemia and any complication was found after adjusting for confounding variables (OR = 2.963, 95%CI: 1.451 - 6.050, P = 0.003), and American society of anesthesia score (ASA score) was significantly associated with any major complication (OR = 2.520, 95%CI: 1.003 - 6.332, P = 0.049). CONCLUSIONS: Radical cystectomy is associated with a high perioperative complications, using the modification of the Clavien system has allowed us to stratify complications during radical cystectomy. Hypoproteinemia is independently associated with any complication in these patients and ASA score was significantly associated with any major complication.

Successful robot-assisted partial nephrectomy for giant renal hilum angiomyolipoma through the retroperitoneal approach: A case report.

BACKGROUND: Giant renal angiomyolipomas (AMLs) may lead to complications including flank pain, hematuria, hypertension, retroperitoneal hemorrhage and even death. Giant AMLs which grow around renal hilar vessels and the ureter are rare. Most previous reports on the treatment of giant renal AMLs have focused on open surgery or a transperitoneal approach, with few studies on the retroperitoneal approach for large AMLs. We here report a case of giant renal hilum AML successfully treated with robot-assisted laparoscopic nephron sparing surgery the retroperitoneal approach, with a one-year follow-up. CASE SUMMARY: A 34-year-old female patient was diagnosed with renal AML 11 years ago and showed no discomfort. The tumor gradually increased in size to a giant AML over the years, which measured 63 mm × 47 mm ×90 mm and was wrapped around the right hilum. Therefore, a robotic laparoscopic partial nephrectomy (LPN) via the retroperitoneal approach was performed. The patient had no serious postoperative complications and was discharged soon after the operation. At the one-year follow-up, the patient's right kidney had recovered well. CONCLUSION: Despite insufficient operating space via the retroperitoneal approach, LPN for giant central renal AMLs can be completed using a well-designed procedure with the assistance of a robotic system.

Construction of a recombinant eukaryotic expression plasmid containing human PDLIM2 gene and its biological activity.

In order to obtain a full-length expression plasmid for human PDLIM2 gene, fragment amplification was used to clone its full-length coding sequence (CDS) region. The amplified PCR product was then digested and inserted into the pMD 18-T vector and subcloned into the pIRES2-EGFP plasmid to form the pIRES2-EGFP-PDLIM2 eukaryotic expression vector. After it was transfected to the bladder cancer cell line, BIU-87, the biological activities of high expression were verified by RT-PCR and Western blotting. Meanwhile the mRNA and protein expressions of p65 were detected. Finally we analyzed the effect of overexpressed PDLIM2 on BIU-87 cell proliferation. In conclusion, a recombinant eukaryotic expression vector pIRES2-EGFP-PDLIM2 containing the complete CDS region of PDLIM2 was successfully constructed. PDLIM2 negatively regulated p65 expression and inhibited BIU-87 cell proliferation. We laid the foundations for further research into the function of the PDLIM2 gene in bladder cancer.

[Transrectal ultrasonography in the etiological diagnosis of male obstructive azoospermia: analysis of 695 cases].

OBJECTIVE: To assess the role of transrectal ultrasonography (TRUS) in the etiological diagnosis of male obstructive azoospermia. METHODS: We retrospectively analyzed the clinical data and TRUS findings of 695 patients with obstructive azoospermia from January 2007 to May 2009. RESULTS: Concerning the etiology of obstructive azoospermia, the main TRUS findings included ejaculatory duct abnormality (29.2%), seminal vesicle abnormality (25.4%) and prostate midline cyst (18.5%). TRUS revealed 203 cases of ejaculatory duct dilation, 177 cases of seminal vesicle abnormality (including 108 with absence or agenesis and 51 with dilation of the seminal vesicle), and 128 cases of prostate midline cyst (including 75 with ejaculatory duct cyst and 39 with Müllerian cyst). Calcification of the verumontanum or ejaculatory duct was suspected to be the causes of obstructive azoospermia in 34 cases. However, no significant etiological abnormality was found in 153 cases. Obvious etiology was shown by TRUS in 78.0% of the patients. CONCLUSION: TRUS can clearly display the structural abnormality of the ejaculatory duct and seminal vesicle, and provide important information on the etiology of male obstructive azoospermia.

Comparison of surgical stress response to laparoscopic and open radical cystectomy.

OBJECTIVES: To prospectively compare stress response to laparoscopic and open radical cystectomy by the measurement of humoral mediators and the incidence of systemic inflammatory response syndrome (SIRS). METHODS: Thirty-eight patients undergoing radical cystectomy were prospectively assessed. Blood samples were obtained from all patients before surgery, during surgery, 72 h after surgery. Serum levels of interleukin (IL)-6 and interferon (IFN)-gamma were measured using an enzyme-linked immunosorbent assay. We also investigated the incidence and duration of SIRS in the two groups. RESULTS: The two groups had comparable perioperative variables except for less estimated blood loss in the laparoscopic group. The IL-6 levels increased during and after surgery in the two groups (P < 0.001). However, the IL-6 levels in the laparoscopic group were significantly lower than those in the open group during and after surgery (P = 0.006, P < 0.001). The incidence of SIRS was 57.1% in the laparoscopic group and 79.2% in the open group (P = 0.149). The mean duration of SIRS was 1.4 days in the laparoscopic group and 2.8 days in the open group (P = 0.032). The IFN-gamma levels decreased, but there was no difference in the two groups over the entire period assessed. Multivariate analysis demonstrated that the group (laparoscopic versus open) was the only influencing factor on the levels of IL-6 and the duration of SIRS. CONCLUSIONS: Our study suggests that the laparoscopic group is markedly less stressful and it has a shorter duration of SIRS than the open group.

Analysis of long-term survival in patients with localized renal cell carcinoma: laparoscopic versus open radical nephrectomy.

OBJECTIVES: To assess the impact of surgical approaches and clinico-pathological parameters on the prognosis of localized renal cell carcinoma (RCC) after laparoscopic radical nephrectomy (LRN) or open radical nephrectomy (ORN). METHODS: Between 2002 and 2008, 336 patients with localized RCC were treated at our two institutions. Of these, 261 were classified as clinical stage pT1 and 75 as pT2, the mean age was 52.3 years. One hundred forty-two patients underwent LRN, and 194 underwent ORN. The medical records of all patients were retrospectively reviewed. RESULTS: The mean follow-up was 44 months (12-84 months). The 3-, 5-, and 7-year cancer-specific survival (CSS) rates of LRN group (96.3, 94.6, and 92.5%, respectively) were equivalent to ORN (97.4, 94.2, and 91.4%, respectively). The mean CSS rates were not significantly different between LRN and ORN groups for either pT1 (82.3 vs. 81.6 months, P = 0.713) or pT2 (69.0 vs. 72.1 months, P = 0.729). Univariate analysis demonstrated significant impact of tumor stage (pT1 vs. pT2, P = 0.002) and tumor grade (G1/2 vs. G3/4, P = 0.045) on CSS. Surgical approach (laparoscopic or open) was not a statistically significant factor on CSS. Multivariate analysis identified that tumor stage was an independent prognostic factor for CSS of localized RCC (P = 0.007). CONCLUSIONS: Based on the long-term follow-up, our evaluation suggests that LRN is oncologically equivalent to ORN for patients with pT1 or pT2 tumors. Among the clinical variables, tumor stage appears to be an independent prognostic factor of CSS of localized RCC patients.

Comparison of hand-assisted laparoscopic and open radical cystectomy for bladder cancer.

PURPOSE: To present our experience of hand-assisted laparoscopic radical cystectomy (HALRC) and compare it with open radical cystectomy (ORC). PATIENTS AND METHODS: During the period between May 2004 and November 2007, 31 patients underwent HALRC and 39 patients underwent ORC for bladder cancer. The patient demographics, intraoperative variables and postoperative outcomes were compared retrospectively. RESULTS: There was no statistically significant difference with regard to age, sex, body mass index, and urinary diversion in the 2 groups. In the HALRC group, 20 patients had stage pT2 or less, 7 had pT3 disease, and 4 had stage pT4 disease. In the ORC group, 29 patients had stage pT2 or less, 5 had pT3 disease, and 5 had stage pT4 disease. All margins in both groups were negative. The HALRC group had decreased blood loss (250.9 vs. 812.8 ml, p < 0.001) and a lower rate of transfusion (9.7 vs. 76.9%, p < 0.001), but similar mean operative time (365.7 vs. 362.6 min, p = 0.862). Time to liquid diet was significantly less in the HALRC group versus the ORC group (4.3 vs. 6.3 days, p < 0.001). The median number of lymph nodes were similar between the HALRC and ORC groups (14 vs. 15, p = 0.377). Six patients developed perioperative complications in the HALRC group and 12 patients had complications in the ORC group (19.4 vs. 30.8%, p = 0.278). Late complications occurred in 3 patients (2 parastomal hernias and 1 ureteroenteral stricture) in the HALRC group. CONCLUSIONS: Compared with ORC, HALRC patients had decreased blood loss, less transfusion requirements, and quicker intestinal recovery. Long-term follow-up in a larger cohort of patients is needed to assess the long-term oncological and functional outcomes.

[Multivariate analysis of recurrence factors for non muscle-invasive bladder urothelial carcinomas].

OBJECTIVE: To discuss the risk factors of recurrent non-muscle-invasive bladder cancer and elucidate its clinical significance. METHODS: The retrospective survival analysis of 161 patients with non-muscle-invasive bladder cancer was performed by Kaplan-Meier method, Log-rank test and COX proportional hazard model. RESULTS: On univariate analysis, the parameters of tumor stage, tumor grade, number of tumors and previous recurrence were significant for tumor recurrence (all P < 0.05). On multivariate analysis of COX proportional hazard model, all the above risk factors remained significant for tumor recurrence. The hazard ratios were as follows: tumor stage (RR = 3.810, P = 0.001), tumor grade (RR = 2.416, P = 0.009), number of tumors (RR = 1.736, P = 0.036) and previous recurrence (RR = 1.810, P = 0.010). CONCLUSION: Tumor stage and tumor grade plays the most important role in tumor recurrence.

Hand-assisted versus pure laparoscopic radical cystectomy: a clinical outcome comparison.

OBJECTIVES: To compare hand-assisted laparoscopic radical cystectomy (HALRC) with the standard laparoscopic radical cystectomy (LRC) in an attempt to delineate their role in bladder cancer treatment. METHODS: We retrospectively analyzed 51 patients who underwent HALRC (HALRC group, 31 cases) or LRC (LRC group, 20 cases). Urinary diversion was performed extracorporeally through the hand port or the incision for specimen retrieval, respectively, in the two groups. Baseline patient characteristics, intraoperative parameters, and postoperative outcomes were evaluated. RESULTS: There was no statistically significant difference in age, sex, body mass index, previous abdominal surgeries, or tumor stage between the two groups. Although the LRC group yielded a significantly smaller incision for urinary diversion than the HALRC group (7.3 cm vs 6.2 cm, P < 0.05), mean operative time, mean estimated blood loss, blood transfusion rate, time to oral intake and complications were similar in the two groups. Hernia formation was observed with increased frequency in the HALRC group. No patients in the HALRC group and only one patient (5%) in the LRC group presented a positive margin. CONCLUSIONS: The HALRC group yielded the same outcomes as the LRC group, except with a larger incision. The hand-assisted approach might be preferred for obese patients or those having multiple previous abdominal surgeries.

[A case-control study of environmental and genetic factors and prostate cancer in Guangdong].

OBJECTIVE: To explore the etiologic relationship of prostate cancer and environmental and genetic polymorphism in southern China METHODS: A hospital-based and 1:1 matched case-control study was conducted. A total of 142 matched pairs of subjects were investigated in this study. The blood samples were collected from 85 cases of prostate cancer and 82 controls of other diseases after informing consent. The CYP1A1, CYP17 and AR genes were analyzed by using the method of PCR, PCR-RFLP. The data were analyzed with conditional logistic regression model. RESULTS: An increased risk of prostate cancer development was observed with the early first spermatorrhea (age < 18) (OR = 2.90, 95% CI: 1.76 - 4.80), early first sexual intercourse (age < or = 24) (OR = 2.38, 95% CI: 1.14 - 4.96), frequent sexual intercourse before 35 year old (OR = 1.80, 95% CI: 1.19 - 2.70), family history of cancer (OR = 2.70, 95% CI: 1.31 - 5.58), more intake of pork (OR =2.27, 95% CI: 1.38 - 3.70). Factors in lowing the risks were the fruit intake and drinking of green tea by OR value at 0.25 (95% CI: 0.08 - 0.75) and 0.52 (95% CI: 0.28 -0.96) respectively. CYP17 A1/A2 and CYP17 A2/A2 genotypes were related with a high risk of prostate cancer and OR values of 1.78 (95% CI: 0.70 - 4.53) and 2.57 (95% CI: 0.91 - 7.25) respectively. Study also showed that there was an interaction between CYP17 polymorphisms and early first spermatorrhea and family cancer history related to the risk of prostate cancer with OR value at 13.35 (95% CI: 1.58 - 113.00) and 4.01 (95% CI: 1.22 - 13.17) respectively. CONCLUSION: Sexual intercourse, dietary intake and family cancer history should be related to prostate cancer occurrence. CYP17 polymorphism might be associated with a high risk of prostate cancer. It suggests that there are multiple environmental and genetic factors to the prostate cancer.

Hand-assisted laparoscopic radical cystectomy and extracorporeal urinary diversion: experience with 31 cases.

OBJECTIVES: To report our techniques and experience with hand-assisted laparoscopic radical cystectomy and extracorporeal urinary diversion for bladder cancer. METHODS: Between May 2004 and November 2007, 31 patients (mean age 61.3 years, range 40-79) underwent hand-assisted laparoscopic radical cystectomy with extracorporeal urinary diversion for bladder cancer. Five patients had previously undergone abdominal surgeries. Data were collected with respect to patient demographics, perioperative outcomes and short-term oncological follow up. RESULTS: Twenty-four patients underwent an ileal conduit and seven patients underwent an orthotopic neobladder. Mean operative time was 365.7 min (range 245 to 530). Estimated blood loss was 250.9 cc (range 100 to 500), with a transfusion rate of 9.7%. Oral liquids were resumed at 4.3 days and the mean hospital stay was 19.7 days. There were no intraoperative complications. Postoperative early complications (within 30 days of surgery) occurred in six patients (19.4%). Two wound infections, one urinary leak, one wound dehiscence, one bowel obstruction and one alimentary tract hemorrhage were all treated conservatively. Late complications occurred in three patients (two parastomal hernias and one ureteroenteric stricture). With a mean follow up of 18 months, 27 patients had no evidence of disease. One patient died because of cancer and one died for unrelated causes. One was alive with local recurrences and one with lung metastasis. CONCLUSIONS: Hand-assisted laparoscopic radical cystectomy is a safe, reproducible and minimally invasive option for bladder cancer patients.

[Holmium laser urethrotomy for male urethral stricture].

OBJECTIVE: To evaluate transurethral holmium laser incision, its safety and effect in the treatment of male urethral stricture. METHODS: Thirty-eight males with urethral stricture were treated by 1045 W holmium laser urethrotomy, 18 with the stricture length shorter than 1.0 cm, 9 between 1.0 cm and 1.5 cm, 7 longer than 1.5 cm , 4 with occlusive stricture and 6 companies with bladder calculus. The average peak urinary flow rate (Q(max)) was (5.6 +/- 2.3) ml/s. RESULTS: Successful surgery was achieved in 36 of the cases, with no complications and the average Q(max) increased to (17.5 +/- 3.4) ml/s. Two cases were converted to open surgery. Thirty-two cases were followed up for 3-18 months, of whom 4 received urethral dilation and 2 underwent a second holmium laser urethrotomy. CONCLUSION: Holmium laser urethrotomy is a safe, effective and minimally invasive therapeutic modality for male urethral stricture.

[Clinical analysis of renal cell carcinoma: report of 271 cases].

OBJECTIVE: To study the diagnosis and treatment of renal cell carcinoma. METHOD: From January 1993 to December 2000 the data of 271 cases of renal cell carcinoma were reviewed. RESULTS: Ultrasonography and CT scanning were still the main diagnostic methods. Surgical operation was performed on 234 patients. Radical nephrectomy was performed on 197 patients (72.6%); Nephron sparing surgery was performed on 19 patients; Metastatic tumor resection was performed on 6 patients and other procedures for 12. The pathological results showed that 137 cases (61.4%) were clear cell carcinoma, 18 cases (8. 1%) of granular cell carcinoma, 32 cases (14. 3%) being combination of the above two varieties, 23 cases (10.3%) of renal papillary adenocarcinoma, 13 cases being renal cell of other types. And 210 cases (77.5%) had been successfully followed up. The 1, 3, 5 and 10 year survival rates were 95.3% (182/191), 88.7% (107/122), 74.7% (56/75) and 32.1% (10/31) respectively. CONCLUSIONS: Ultrasonography is the first select examination method of detecting of renal cell carcinoma, and CT scanning is the most valuable diagnostic mean. Early diagnosis and prompt radical nephrectomy or nephron sparing nephrectomy are the critical points for achieving long-term survivals of patients with renal cell carcinoma.

[The clinico-pathological significance of protein expression of PAK1 in bladder transitional cell carcinoma].

OBJECTIVE: To investigate the protein expression of p21-activated kinase 1 gene (PAK1) in bladder transitional cell carcinoma (BTCC) and its clinico-pathological significance. METHODS: Immunohistochemistry and TUNEL were used, in combination with tissue microarray technique, to examine the protein expression of PAK1 and status of cell apoptosis in 100 BTCC tissue specimens obtained during operation and 30 specimens of adjacent normal bladder mucosa. RESULTS: All adjacent normal bladder mucosa specimens were negative in PAK1 protein expression or only with a low-level expression of PAK1 protein, while 58% of the BTCC specimens showed over-expression of PAK1. PAK1 expression was significantly associated with tumor pathological grade and tumor size (both P < 0.05). The PAK1 overexpression rate of the poorly-differentiated BTCC specimens (at the G3 stage) was 78%, significantly higher than that of the well-differentiated specimens (at the stage G1/G2, 47%, P = 0.05). The PAK1 overexpression rate of the large-sized BTCC specimens (>or= 3 cm in diameter) was 73%, significantly higher than that of the small-sized BTCC specimens (< 3 cm in diameter, P = 0.034). The PAK1 protein expression was negatively correlated with the apoptotic index of the cells (P < 0.05). CONCLUSION: Overexpression of PAK1 protein may via its anti-apoptotic function to play an important role in the development and progression of BTCC. Overexpression of PAK1 in BTCC is associated closely with tumor malignant histological phenotype and it may be used as a molecular marker to predicate the malignant potential of BTCC.

[siRNA knocks down the expression of AR gene in PC3 cell lines].

OBJECTIVE: To observe the suppression of the expression of androgen receptor (AR) gene in PC3 cells after AR-specific siRNAs transfection, and to search for the siRNA (s) with the greatest suppressing efficiency. METHODS: Five AR-specific siRNAs were selected, RNAi expression vectors were constructed and transfected into PC3 cells, and the AR expression was detected by real time FQ-PCR and Western blot. A nonsense small RNA was set as negative control. RESULTS: Compared with the control group, the AR expression decreased in various degrees in the 5 experimental groups (P < 0.05), and siRNA1, siRNA4 and siRNA5 showed the greatest suppressing efficiency as compared with the other experimental groups, with statistically significant difference (P < 0.05). CONCLUSION: The AR-specific siRNAs could suppress the endogenous expression of target gene. Three siRNAs with great suppressing efficiency were identified and the expression vectors were constructed successfully. It can be applied in the future researches in vivo.

[The clinical significance of expression and amplification of FGF3 in bladder transitional cell carcinoma].

OBJECTIVE: To investigate the amplification and expression of FGF3 in bladder transitional cell carcinoma (BTCC) and its clinical significance. METHODS: Immunohistochemistry (IHC) and Fluorescence In Situ Hybridization (FISH) methods were used to examine the protein expression and amplification of FGF3 in a tissue microarray (TMA) of 100 BTCCs and 30 adjacent normal bladder mucosas, so as to analyze their correlation and association with patient's clinico-pathological features. RESULTS: In this study, none of the normal bladder mucosas were detected FGF3 positivity, while in 89 informative BTCCs, 20 (22%) cases were observed positive expression of FGF3 protein, and it was significantly more frequently to occur in BTCCs of poor-differentiation (Grade 3), later clinical stage (T2-4) and tumor in >or= 3 cm in diameter (P < 0.05). In FISH study, 10 of the 63 (16%) informative BTCCs were observed amplification of FGF3 and it was significantly associated with BTCC's tumor size and clinical stage (P < 0.05). In addition, 10 BTCCs with amplification of FGF3 in this study were all detected positive expression of FGF3 protein, while in the remaining 53 BTCCs without amplification of FGF3, only 3 (6%) cases were observed FGF3 protein positivity. CONCLUSION: The up-regulated expression of FGF3 in BTCC was associated closely with tumor's malignant clinical phenotypes, and it might be involved in the malignant progression of parts of BTCC. The amplification of FGF3 gene might be a predominant mechanism of increased expression of FGF3 protein in BTCC.