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Objective To study the expression of long non-coding RNA (lncRNA)-urothelial carcinoma associated 1 (UCA1) and miR-34b in bladder cancer and its correlation to the clinicopathologic features of bladder cancer.Methods Between January 2011 and October 2012,the expression of UCA1 and miR-34b in 5 bladder cancer cell lines (T24,BIU-87,EJ,T24-MMC,T24-ADM) and 1 normal bladder cell lines (SV-HUC-1) were measured by real-time reverse transcription-polymerase chain reaction (RTPCR).Meanwhile,the 56 bladder cancer specimens and paraneoplastic normal bladder tissues,which diagnosed by pathology were collected from bladder cancer patients undergoing radical resection of bladder.Among them,41 cases were male and 15 cases were female.The mean age was (68.4 ± 7.5)years old,range 52 to 78 years.43 cases were older than 65 years old,and 13 cases were less than 65 years old.The pathological classification included non muscle-invasive bladder cancer (NMIBC) 18 cases,muscle-invasive bladder cancer 38 cases;low grade papillary urothelial carcinoma 22 cases,high grade papillary urothelial carcinoma 34 cases;12 cases were primary lesion,the other 44 cases were diagnosed as tumor recurrence.Real-time RT-PCR was performed to analyze the expression of UCA1 and miR-34b.Results The relative expression levels of UCA1 in the normal bladder cell lines (SV-HUC-1) and 5 bladder cancer cell lines (T24,BIU-87,EJ,T24-MMC and T24-ADM) were (0.0675 ± 0.0133),(0.2934 ± 0.0531),(0.4246 ± 0.0650),(0.4206 ± 0.0826),(0.6472 ± 0.0875) and (0.7165 ± 0.1032),respectively (P ＜ 0.05).Moreover,the expression levels of UCA1 were up-regulated in 2 drug resistant bladder cancer cells lines T24-MMC (0.6472 ± 0.0875)and T24-ADM (0.7165 ± 0.1032),as compared with the T24 bladder cancer lines (0.2934 ± 0.0531),respectively (P ＜ 0.05).However,the expression levels of miR-34b in 5 bladder cancer cell lines [T24 (0.1600 ± 0.0455),BIU-87 (0.1720 ± 0.0658),EJ (0.1150 ± 0.0352),T24-MMC(0.0576 ± 0.0087),T24-ADM (0.0510 ± 0.0125)] were decreased (P ＜ 0.05),as compared with normal bladder cell lines SV-HUC-1 (0.6384 ± 0.1083).Moreover,the expression levels of miR-34b were down-regulated in 2 drug resistant bladder cancer cells lines T24-MMC (0.0576 ± 0.0087) and T24-ADM(0.0510 ± 0.0125),as compared with the T24 bladder cancer lines T24 (0.1600 ± 0.0455),respectively (P ＜ 0.05).The relative expression levels of UCA1 and miR-34b in bladder cancer tissues and paraneoplastic normal bladder tissues were (0.4225 ± 0.0714) vs.(0.0532 ± 0.0192) and (0.0340 ± 0.0134)vs.(0.5643 ±0.0616),respectively (P ＜0.05).Statistical correlation analysis showed that UCA1 to be significantly negative correlated with miR-34b in bladder cancer specimens(r =-0.54,P ＜ 0.05).The high level of UCA1 and low level of miR-34b were significantly correlated with tumor malignant grade,invasiveness and recurrence.The 3-year overall survival rate (OS) in UCA1 (+)/miR-34b(-) group (27.6％) were significantly worse compared with non UCA1 (+)/miR-34b (-) group (73.7％).Conclusion High expression of UCA1 and low expression of miR-34b were associated with the occurrence and development of bladder cancer.
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Objective To compare the clinical efficacy of laparoscopic surgery and open surgery for intraperitoneal bladder rupture.Methods From January 2004 to August 2013,the clinical data and therapeutic methods of 50 patients with intraperitoneal bladder rupture were retrospectively reviewed,inclu-ding 26 cases of laparoscopic surgery and 24 cases of open surgery.The operative time,intraoperative blood loss,postoperative intestinal recovery,hospital stay,analgesic use rate and complication ratio were com-pared between the two groups.Results All surgeries were successfully performed.There were significant differences in intraoperative blood loss [(54.24 ±5.38)ml vs(89.35 ±12.17)ml],intestinal recovery time [(23.24 ±2.39)h vs(38.42 ±6.98)h],hospital stay [(4.64 ±1.42)d vs(7.04 ±1.29)d]and analgesic use rate [38.64%(10 /26)vs 75.00%(18 /24)]between laparoscopy group and open surgery group,respectively(P 0.05).Conclusion Laparoscopic treatment of intraperitoneal bladder rupture has the advantages of mini-invasion and rapid re-covery compared with traditional open surgery.
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Objective To investigate the causes and treatment of familial neurohypophyseal diabetes insipidus with hydronephrosis.Methods A retrospective analysis was conducted in 6 cases (5 males and 1 female,aged 11 to 53 years) of familial neurohypophyseal diabetes insipidus with hydronephrosis treated in our institute from June 2009 to December 2010.All cases had polydipsia and polyuria since their childhood.The daily output of urine ranged from 5,290 to 15,040 ml.The urine specific gravity was less than 1.005.The water deprivation and vasopressin injection test showed positive results,and MRI showed that the shape and size of pituitary gland were in normal range.Ultrasound and IVU showed that all cases had hydronephrosis.Five adult cases were administered with Desmopressin 0.2 mg three times a day,and 1 juvenile patient given half dosage of Desmopressin as in adult.The case No.1 underwent percutaneous nephrostomy and bilateral ureteral reimplantation.Case No.2 received urethral catheterization for 5 days and Tamsulosin.Three cases with urinary tract infection were given antibiotics on the base of urine culture and antibiotic sensitivity test results.Follow-up was undertaken every 3 mon for the duration of 18-36 mon.Results In 6 cases,polydipsia and polyuria were significantly improved after the treatment.Daily urine output dropped to 6000 ml in 5 adult cases and decreased to 2000 ml in the juvenile case.The flank sore of case No.1 was relieved after percutaneous nephrostomy,and hydronephrosis improved 6 mon after bilateral ureteral reimplantation.The residual urine volume of case No.2 was reduced to 40 ml,and no recurrence was observed after anti-infection therapy.During the follow-up,6 cases showed relieved hydronephrosis and no recurrent infection.Conclusions It is of important to reduce the urine volume for the treatment of familial neurohypophyseal diabetes insipidus with hydronephrosis.Early diagnosis and treatment of the diseases is crucial for the improvement of renal function.
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Fournier's gangrene (FG) is an extremely aggressive and rapidly progressive polymicrobial soft tissue infection of the perineum, anal area or genitalial regions with a high mortality rate. The objectives of this study were to share our experience with the management of this serious infectious disease over the last 15 years. This retrospective study examined 24 patients diagnosed as having FG who were admitted to our hospital between March 1996 and December 2011. The gender, age, etiology, predisposing factors, laboratory findings, treatment modality, hospitalization time and spread of gangrene of the subjects were all recorded and analyzed. The results showed that the mean age of the patients was 48.33 years, the male-to-female ratio was 5:1 and the mortality rate was 20.8% (5/24). The most common predisposing factor was diabetes mellitus in 10 patients (41.6%), followed by alcohol abuse, obesity, neoplasms and immunosuppression. The most common etiology was peri-anal and peri-rectal abscesses (45.8%), followed by lesions of urogenital origin (33.3%) and cutaneous (8.3%) origin. No local pathologies could be identified in 3 (12.5%) patients. The most commonly isolated microorganisms were Escherichia coli (62.5%), followed by Enterococcus, Pseudomonas aeruginosa and Staphylococcus aureus. The median admission Fournier's gangrene severity index (FGSI) score for survivors was 5.63±1.89 against 13.6±3.64 for non-survivors which was designed for predicting the disease severity in the series. Early diagnosis and immediate extensive surgical debridement were significant prognostic factors in the management of Fournier gangrene. Individualized reconstructive modalities for wound coverage were useful in that they repaired the tissue defect and improved the quality of life. We are led to conclude that Fournier's gangrene is a severe condition with a high mortality. The Fournier's gangrene severity index (FGSI) score at admission serves as a good predictor for the disease severity. Early diagnosis, surgical debridement and aggressive fluid therapy are significant prognostic factors in the management of Fournier gangrene. Individualized reconstructive surgery modalities for wound coverage are useful to correct the tissue defect and improve the quality of life.
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This study presented our experience in the treatment of testicular torsion, which may help achieve early diagnosis and improve therapeutic effects. A retrospective analysis was conducted in 71 patients with testicular torsion who were treated in our hospital from October 2007 to April 2011. The age of the patients ranged from 16 days to 34 years. All the patients had unilateral testicular torsion, which took place on the left side in 43 cases and on the right side in 28 cases. The course of the disease varied between three hours to 30 days. Post-operative follow-up was conducted until October 2011. Items examined included signs and symptoms at their first clinical visit, ultrasound findings, treatment in emergency surgery, and post-operative follow-up. In this study, the 71 patients were diagnosed with testicular torsion by color Doppler sonography, 7 had testicular fixation, 63 patients received orchiectomy, while 1 patient did not undergo surgery due to pressure from family members. Post-operative follow-up showed that the one patient's testicle, which had been reserved, atrophied, while all the other survived. No recurrence was found during the follow-up visits. It is concluded that an early diagnosis and surgery is important in improving the survival rate of testicular torsion, and the diagnosis and treatment by the first attending clinician is of critical importance.
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The effect of Smac gene on the TRAIL-induced apoptosis of the prostate cancer cell line PC-3 and the molecular mechanism were investigated. The Smac gene was transfected into PC-3 cells under the induction of liposome. The intrinsic Smac gene expression was detected by Western blotting. After treatment with TRAIL as an apoptosis inducer, in vitro cell growth activity was assayed by MTT colorimetry. The apoptosis rate of PC-3 cells was determined by annexin V-FITC and propidium iodide staining flow cytometry. The expression of cellular XIAP and caspase-3 genes was examined by Western blotting. Smac-transfected cells (PC-3/Smac group) had significantly increased Smac protein level as compared with PC-3 controls (P<0.01). After induction with 100-200 ng/mL TRAIL for 12-36 h, cellular proliferation rate in PC-3/Smac group was significantly lower than in PC-3 controls (P<0.05). After induction with 100 ng/mL TRAIL for 24 h, the apoptosis rate in PC-3/Smac group was significantly enhanced as compared with that of PC-3 controls (P<0.05). Accordingly, the XIAP expression level was down-regulated significantly (P<0.05) and caspase-3 subunit P20 was up-regulated significantly (P<0.05). It is suggested that the over-expression of cellular Smac can inhibit inhibitor of apoptosis proteins (IAPs), enhance caspases activity and the apoptosis rate of PC-3 cells induced by TRAIL, which may provide a useful experimental basis for prostate cancer therapy.
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This study presented our experience in the treatment of testicular torsion, which may help achieve early diagnosis and improve therapeutic effects. A retrospective analysis was conducted in 71 patients with testicular torsion who were treated in our hospital from October 2007 to April 2011. The age of the patients ranged from 16 days to 34 years. All the patients had unilateral testicular torsion, which took place on the left side in 43 cases and on the right side in 28 cases. The course of the disease varied between three hours to 30 days. Post-operative follow-up was conducted until October 2011. Items examined included signs and symptoms at their first clinical visit, ultrasound findings, treatment in emergency surgery, and post-operative follow-up. In this study, the 71 patients were diagnosed with testicular torsion by color Doppler sonography, 7 had testicular fixation, 63 patients received orchiectomy, while 1 patient did not undergo surgery due to pressure from family members. Post-operative follow-up showed that the one patient's testicle, which had been reserved, atrophied, while all the other survived. No recurrence was found during the follow-up visits. It is concluded that an early diagnosis and surgery is important in improving the survival rate of testicular torsion, and the diagnosis and treatment by the first attending clinician is of critical importance.
Subject(s)
Adolescent , Adult , Humans , Male , Young Adult , Emergency Treatment , Methods , Postoperative Period , Testis , General SurgeryABSTRACT
Fournier's gangrene (FG) is an extremely aggressive and rapidly progressive polymicrobial soft tissue infection of the perineum, anal area or genitalial regions with a high mortality rate. The objectives of this study were to share our experience with the management of this serious infectious disease over the last 15 years. This retrospective study examined 24 patients diagnosed as having FG who were admitted to our hospital between March 1996 and December 2011. The gender, age, etiology, predisposing factors, laboratory findings, treatment modality, hospitalization time and spread of gangrene of the subjects were all recorded and analyzed. The results showed that the mean age of the patients was 48.33 years, the male-to-female ratio was 5:1 and the mortality rate was 20.8% (5/24). The most common predisposing factor was diabetes mellitus in 10 patients (41.6%), followed by alcohol abuse, obesity, neoplasms and immunosuppression. The most common etiology was peri-anal and peri-rectal abscesses (45.8%), followed by lesions of urogenital origin (33.3%) and cutaneous (8.3%) origin. No local pathologies could be identified in 3 (12.5%) patients. The most commonly isolated microorganisms were Escherichia coli (62.5%), followed by Enterococcus, Pseudomonas aeruginosa and Staphylococcus aureus. The median admission Fournier's gangrene severity index (FGSI) score for survivors was 5.63±1.89 against 13.6±3.64 for non-survivors which was designed for predicting the disease severity in the series. Early diagnosis and immediate extensive surgical debridement were significant prognostic factors in the management of Fournier gangrene. Individualized reconstructive modalities for wound coverage were useful in that they repaired the tissue defect and improved the quality of life. We are led to conclude that Fournier's gangrene is a severe condition with a high mortality. The Fournier's gangrene severity index (FGSI) score at admission serves as a good predictor for the disease severity. Early diagnosis, surgical debridement and aggressive fluid therapy are significant prognostic factors in the management of Fournier gangrene. Individualized reconstructive surgery modalities for wound coverage are useful to correct the tissue defect and improve the quality of life.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Fournier Gangrene , Diagnosis , Pathology , Retrospective StudiesABSTRACT
The growth inhibition and pro-apoptosis effects of dracorhodin perchlorate on human prostate cancer PC-3 cell line were examined. After administration of 10-80 μmol/L dracorhodin perchlorate for 12-48 h, cell viability of PC-3 cells was measured by MTT colorimetry. Cell proliferation ability was detected by colony formation assay. Cellular apoptosis was inspected by acridine orange-ethidium bromide fluorescent staining, Hoechst 33258 fluorescent staining, and flow cytometry (FCM) with annexin V-FITC/propidium iodide dual staining. The results showed that dracorhodin perchlorate inhibited the growth of PC-3 in a dose- and time-dependent manner. IC50 of dracorhodin perchlorate on PC-3 cells at 24 h was 40.18 μmol/L. Cell clone formation rate was decreased by 86% after treatment with 20 μmol/L of dracorhodin perchlorate. Some cells presented the characteristic apoptotic changes. The cellular apoptotic rates induced by 10-40 μmol/L dracorhodin perchlorate for 24 h were 8.43% to 47.71% respectively. It was concluded that dracorhodin perchlorate significantly inhibited the growth of PC-3 cells by suppressing proliferation and inducing apoptosis of the cells.
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In our previous study, we identified a novel testis-specific expressed gene 2 (TSEG-2) from mouse testis. To further investigate its functions, 35 male Balb/c mice (8 weeks old) were divided into cryptorchidism group (n=20), sham group (n=10), and control group (n=5). In cryptorchidism group, the right testes were anchored to the inner lateral abdominal wall. In situ hybridization (ISH) was applied to measure the localization of TSEG-2 in mouse testis. Real-time quantitative PCR was performed to detect the expression of TSEG-2 gene. Meanwhile, under the mediation of polyethylenimine (PEI), the recombinant vector pEGFP-TSEG-2 (n=5) or empty vector (mock, n=5) was transfected into the testis of male mice. The transfection efficiencies were measured under a fluorescence microscope. The apoptosis of spermatogenic cells was detected by terminal deoxynuleotidyl-mediated nick end labeling (TUNEL). The results showed that TSEG-2 was expressed in convoluted seminiferous tubules, more precisely, in spermatogonia and spermatocytes. As compared with sham and control groups, the TSEG-2 transcription was significantly enhanced (P<0.05) and was correlated with apoptosis of spermatogenic cells in cryptorchid testes (P<0.05). PEI was efficient in mediating transfection of TSEG-2 into seminiferous tubules of testis. One week post-transfection, intratesticular injection of TSEG-2 resulted in increased apoptosis of spermatogenic cells in vivo (P<0.05). These results indicate that TSEG-2 may participate in the apoptosis of spermatogenic cells and the pathogenesis of cryptorchidism.
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Objective To evaluate the efficacy and safety of α1-adrenergic blockers for treating young men with primary bladder neck obstruction (PBNO). Methods A retrospective review was done of the presenting symptoms and videourodynamic findings of 22 young men younger than 35 years with PBNO. Mean age was 28 years (range 18 to 35). The presenting symptoms were hesitancy in 21 (95%), weak stream in 17(77%), frequency in 16(73%), urgency in 8(36%) and pelvic pain in 6 (27%). Mean symptom duration was 28(3-62)months. A dose of 4 mg Doxazosion was adminis-tered for at least 6 months. International prostate symptom score(IPSS), Quality of life(QOL), uro-flowmetry, post-void residual urine and blood pressure were assessed before and 6 months after medi-cation. Improved urine flow was defined as at least 3 ml. per second increase in the maximum flow rate. Improved symptom was defined as more than a 40% decrease in IPSS. Successful treatment was defined as improved in urine flow and symptoms. Results Follow-up data were available for 21 of 22 patients. The medication period was 8.7±2.5 months and follow-up duration was 12.3±4.9 months. Mean Ⅰ-PSS decreased from 16.9±3.7 to 10.7±4.5. Mean QOL decreased from 4.3±1.2 to 2.5±1.0. Mean maximum flow rate increased from (9.8±3.5)ml to (14.9±3.6)ml. per second. Mean post-void residual urine decreased from (78.2±35.6)ml to (46.5±19.4)ml. There were significant differences(P<0.01). Treatment was successful in 14 patients (67%). Drug tolerability was good. Mean blood pressure was (110.0±7.9)/ (75.0±5.9)mm Hg and (107.0±8.7)/(72.0±7.1)mm Hg before medicine therapy and after 6 months medication(P>0.05). Conclusions Videourody-namics is the diagnostic gold standard of PBNO. In our experience α1-adrenergic blockers are clinically effective therapy and safety for PBNO and have been well tolerated in young male patients.
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AIM:To investigate the activation and inactivation of nuclear factor kappa B(NF-κB)when tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)is applied to induce the apoptosis of androgen-independent prostate cancer cell line PC-3M.METHODS:After the treatment of TRAIL or LPS at different doses,we tested the nuclear translocation of NF-κB by cell immunohistochemical staining and electrophoretie mobility shift assay(EMSA),and evaluated the level of IκB by RT-PCR under pyrrolidine dithiocarbamate(PDTC)treatment.RESULTS:EMSA and cell immunohistochemical analysis showed that the translocation of NF-κB was significantly activated when PC-3M cells were treated with TRAIL or LPS(P＜0.05).The pretreatment of PDTC upregulated the expression of IκB and blocked the nuclear translocation of NF-κB.CONCLUSION:TRAIL remarkably stimulates the activation of nuclear NF-κB in androgen-independent prostate cancer cells.On the other hand,the translocation of NF-κB can be significantly and efficiently inhibited in PC-3M cells by pretreatment with PDTC.The increased expression of IκB might be a clue for this inhibition,which means the possible way to enhance the effect of TRAIL in the apoptosis of prostate cancer cells.
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Objective To investigate the effect of recombinant plasmid pshRNA-DNMT3b on expression of DNMT3b mRNA and protein and on the proliferation of bladder cancer T24 cells,and research the function of DNMT3b in the process of bladder tumor formation.Methods There were three groups in this study,which are blank controller,HK and pshRNA-DNMT3b(24h,48h,72h),respectively.T24 cells were cultured routinely and transfected by the recombinant plasmids with lipfectamine 2000.The cells were detected by methods of RT-PCR,western blot and MTT.The varying level of DNMT3b mRNA and expression protein,and the conditions of cellular survival rate were observed.Results The recombinant plasmids were successfully transfected into T24 cell lines.The grey valHe of RT-PCR elctrophoretogram was analyzed by the software of Gel-pro analyzer,the rate of blank controller,HK and pshRNA-DNMT3b(24h,48h,72h),was (99.56±1.24)%,(99.12±1.35)%,(75.77±1.42)%,(44.69±1.05)%and(20.52±0.89)%,respectively.The analytical resuit of western blot image was(99.43±1.28)%,(98.90±1.31)%,(67.83±1.02)%,(43.43±1.05)%and(21.92±0.89)%.There was no statistically difference in survival between blank control and HK(P＞0.05).The group of pshRNA-DNMT3b and other two groups had statistical difference only at the 72th hour and the cell inhibitory growth rate only increase 0.45%.Conclusions The recombinant ptasmid pshRNA-DNMT3b can inhibit the expression of mRNA and protein of DNMT3b effectively.However,it has slight function on inhibiting cell proliferation.
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The present study was aimed at finding an effective method to isolate and purify the subtype of type A spermatogonial stem cells (SSCs) in juvenile rats. Testes from 9-days-old rats were used to isolate germ cells by using two-step enzymatic digestion. The expression of c-kit in the testes of the rats was immunohistochemically detected. After isolation, cell suspension was enriched further by discontinuous density gradient centrifugation. Then type A1-A4 spermatogonia was isolated from the purified spermatogonia with c-kit as the marker by using fluorescence-activated cell sorting (FACS). Electron microscopy was used to observe their ultrastructure. Finally, highly purified and viable subtype of SSCs was obtained. Cells separation with discontinuous density gradient centrifugation significantly increased the concentration of c-kit positive cells [(18.65+/-1.69)% after the centrifugation versus (3.16+/-0.84)% before the centrifugation, P<0.01]. Furthermore, the recovery and viability were also high [(65.9+/-1.24)% and (85.6+/-1.14)%]. It is concluded that FACS with c-kit as the marker in combination with discontinuous density gradient centrifugation can well enrich type A1-A4 spermatogonia from the testes of 9-days-old rats.
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It has been suggested that progression of bladder transitional cell cancer (BTCC) may be regulated at the molecular level by a typical pattern of expression of genes involved in apoptosis. Recently Livin, belonging to the inhibitors of apoptosis (IAP) family, has been found to be expressed in most solid tumors, where its expression is suggested to have clinical significance. In order to explore the significance of Livin expression in the development of BTCC, immunohistochemistry and RT-QPCR were used to detect the expression of Livin mRNA in tumor tissues and adjacent normal tissues of 30 cases of BTCC. The results showed that the positive rate of Livin expression in adjacent normal tissues and tumor tissues was 0 and 60% (18/30) respectively. The-DeltaDeltaCT value of Livin in BTCC tissues was 8.0454 (7.4264-8.6644) times of that in adjacent normal tissues. The expression of Livin mRNA had no correlation with tumor pathological grades and clinical stages. It was suggested that there was weak expression of Livin mRNA in adjacent normal tissues, but strong in tumor tissues.
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The expression of survivin, a member of inhibitor of apoptosis (IAP) family, was examined in bladder transitional cell cancer (BTCC) tissue and adjacent normal tissues to examine its clinical implication in the development of BTCC. Thirty specimens of bladder cancer were detected for the expression of survivin by using immunohistochemistry and real-time quantitative reverse transcription polymerase chain reaction (RT-QPCR) in BTCC tissue and adjacent normal tissues. Our results showed that the positive rate of survivin immunostaining specimen were 0 and 60% (18/30) in the adjacent normal tissues, bladder cancer, respectively. The-DeltaDeltaCT value of survivin in bladder cancer tissue was 10.2829 (9.0034-11.5624) times that in the adjacent normal tissues. The expressions of survivin were correlated with the pathological grades of tumor and clinical stages. It is concluded that there was only weak expression of survivin mRNA in the adjacent normal tissues, but the expression of survivin mRNA in bladder cancer tissue was much higher than that in the adjacent normal tissues and the expression of survivin was correlated with pathological grades and clinical stages of tumor.
Subject(s)
Carcinoma, Transitional Cell/metabolism , Carcinoma, Transitional Cell/pathology , Microtubule-Associated Proteins/genetics , Microtubule-Associated Proteins/metabolism , Prognosis , RNA, Messenger/genetics , RNA, Messenger/metabolism , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , Urinary Bladder Neoplasms/metabolism , Urinary Bladder Neoplasms/pathologyABSTRACT
Connexin-43 (Cx43) expression in prostate cancer (PCa) cells and the potency of gap junctional intercellular communication (GJIC) in the cells were investigated, with an attempt to elucidate the reason why the so-called "bystander effect" mediated by thymidine kinase (TK) suicide gene therapy on PCa cells is not of significance and to explore the role of GJIC in PCa carcinogenesis. mRNA and protein expression of Cx43 in a PCa cell line PC-3m was detected by reverse-transcription polymerase chain reaction (RT-PCR) and strapt-avidin-biotin-enzyme complex (SABC) immunohistochemical staining, and inherent GJIC of PC-3m cells was assayed by scrape-loading and dye transfer (SLDT) assay. The expression of Cx43 in human normal and malignant prostate tissues was determined by SABC immunohistochemistry as well. It was found that Cx43 mRNA and protein expression in PC-3m cells was slightly reduced as compared with positive controls and the location of Cx43 protein was aberrant in cytoplasm rather than on membrane. Assessment of paraffin sections demonstrated that the expression of Cx43 protein in PCa cells was abnormally located and markedly diminished as compared with normal prostatic epithelial ones, displaying a negative correlation to the pathological grade (chi2=4.025, P<0.05). Additionally, capacity of inherent GJIC in PC-3m cells was disrupted, which was semi-quantified as (+) or (-). It was indicated that both down-regulated expression of Cx43 mRNA and aberrant location of Cx43 protein participated in the mechanisms leading to deficient GJIC in PC-3m cells. Lack of efficient GJIC is a molecular event, which may contribute not only to limited extent of "bystander effect", but also to initiation and progression of prostatic neoplasm.
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The expression of X-linked inhibitor of apoptosis protein (XIAP) gene and its effect on chemotherapeutic sensitivity in bladder carcinoma was explored. By using immunohistochemistry, the expression of XIAP was detected in 47 bladder carcinomas and 5 normal bladder tissues. The XIAP gene was transfected into bladder cancer cell line T24 by liposome and the positive clone was screened by G418. Cellular XIAP mRNA level was detected by RT-PCR. Low-dose mitomycin C was administered to induce the apoptosis of T24 cells. The in vitro growth of bladder carcinoma cells was analyzed by MTT colorimetry, and the apoptosis rate was assayed by TUNEL methods. It was found XIAP was moderately expressed in bladder carcinomas with the the positive rate being 78.73% (37/47), but the positive rate was not correlated with carcinoma stages and grades (P<0.05). XIAP mRNA level in transfected T24 cells was significantly increased by 3.8 times as compared with that in the cells not transfected with XIAP. After treatment with low-dose mitomycin C (0.005 and 0.05 mg/mL), the growth rate in XIAP no-transfected control group was increased by (11.60+/-0.25)% and (16.51+/-0.87)% (P<0.05), and the apoptosis rate was decreased by (10.1+/-0.2)% and (11.9+/-0.2%) (P<0.05) respectively as compared with XIAP transfected group. It was concluded that XIAP was expressed in most of bladder carcinoma samples. Overexpression of XIAP in T24 could significantly reduce the MMC-induced apoptosis of bladder carcinoma, suggesting its effect on the chemotherapeutic sensitivity of T24 cells.
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The expression of X-linked inhibitor of apoptosis protein (XIAP) gene and its effect on chemotherapeutic sensitivity in bladder carcinoma was explored. By using immunohistochemistry,the expression of XIAP was detected in 47 bladder carcinomas and 5 normal bladder tissues. The XIAP gene was transfected into bladder cancer cell line T24 by liposome and the positive clone was screened by G418. Cellular XIAP mRNA level was detected by RT-PCR. Low-dose mitocycin C was administered to induce the apoptosis of T24 cells. The in vitro growth of bladder carcinoma cells was analyzed by MTT colorimetry, and the apoptosis rate was assayed by TUNEL methods. It was found XIAP was moderately expressed in bladder carcinomas with the the positive rate being 78.73% (37/47), but the positive rate was not correlated with carcinoma stages and grades (P＜0.05). XIAP mRNA level in transfected T24 cells was significantly increased by 3.8 times as compared with that in the cells not transfected with XIAP. After treatment with low-dose mitomycin C (0.005 and 0.05 mg/mL), the growth rate in XIAP no-transfected control group was increased by (11.60±0.25)% and (16.51±0.87)% (P＜0.05), and the apoptosis rate was decreased by (10.1±0.2)% and (11.9±0.2%) (P＜0.05) respectively as compared with XIAP transfected group. It was concluded that XIAP was expressed in most of bladder carcimoma samples. Overexpression of XIAP in T24 could significantly reduce the MMC-induced apoptosis of bladder carcinoma, suggesting its effect on the chemotherapeutic sensitivity of T24 cells.
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Connexin-43 (Cx43) expression in prostate cancer (PCa) cells and the potency of gap junctional intercellular communication (GJIC) in the cells were investigated, with an attempt to elucidate the reason why the so-called "bystander effect" mediated by thymidine kinase (TK) suicide gene therapy on PCa cells is not of significance and to explore the role of GJIC in PCa carcinogenesis.mRNA and protein expression of Cx43 in a PCa cell line PC-3m was detected by reverse-transcription polymerase chain reaction (RT-PCR) and strapt-avidin-biotin-enzyme complex (SABC) immunohistochemical staining, and inherent GJIC of PC-3m cells was assayed by scrape-loading and dye transfer (SLDT) assay. The expression of Cx43 in human normal and malignant prostate tissues was determined by SABC immunohistochemistry as well. It was found that Cx43 mRNA and protein expression in PC-3m cells was slightly reduced as compared with positive controls and the location of Cx43 protein was aberrant in cytoplasm rather than on membrane. Assessment of paraffin sections demonstrated that the expression of Cx43 protein in PCa cells was abnormally located and markedly diminished as compared with normal prostatic epithelial ones, displaying a negative correlation to the pathological grade (χ2=4.025, P＜0.05). Additionally, capacity of inherent GJIC in PC-3m cells was disrupted, which was semi-quantified as (+) or (-). It was indicated that both down-regulated expression of Cx43 mRNA and aberrant location of Cx43 protein participated in the mechanisms leading to deficient GJIC in PC-3m cells. Lack of efficient GJIC is a molecular event, which may contribute not only to limited extent of "bystander effect", but also to initiation and progression of prostatic neoplasm.