RESUMO
Objective: To explore the effect and mechanism of Casticin (CAS) on the proliferation, migration and invasion of bladder cancer T24 cells. Methods: T24 cells were cultured in vitro and divided into control group, 5, 10, 20 μmol/L CAS groups, si-NC group, si-TM7SF4 group, CAS+ pcDNA group and CAS+ pcDNA-TM7SF4 group. Cell counting kit-8 (CCK-8) was used to detect cell proliferation; Transwell was used to detect cell migration and invasion; western blot was used to detect the protein expressions of cyclin D1, p21, MMP-2, MMP-9 and TM7SF4, and real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to detect the expression of TM7SF4 mRNA. Results: The inhibition rates of T24 cells in the 5, 10, 20 μmol/L CAS groups were (17.68±1.41)%, (33.54±3.16)% and (61.44±5.50)%, respectively, higher than (0.00±0.00)% of the control group (P<0.001), but the numbers of migration and invasion were 72.83±5.66, 59.13±4.27, 41.25±3.22 and 55.83±5.15, 42.19±3.06, 31.13±3.22, respectively, lower than 86.11±5.16 and 68.82±5.29 of the control group (P<0.001). The protein expression levels of cyclin D1, MMP-2, MMP-9, TM7SF4 and the expression levels of TM7SF4 mRNA in the 5, 10, and 20 μmol/L CAS groups were lower than the control group (P<0.001). However, the protein expression levels of p21 were 0.37±0.03, 0.51±0.04, and 0.66±0.06, respectively, higher than 0.25±0.03 in the control group (P<0.001). The inhibition rate of T24 cells in the si-TM7SF4 group was (50.35±4.67)%, higher than (6.31±0.58)% in the si-NC group (P<0.001), but the numbers of migration and invasion were 53.51±4.18 and 42.92±3.81, lower than 85.26±4.99 and 67.93±4.64 of the si-NC group (P<0.001). The protein expression levels of TM7SF4, CyclinD1, MMP-2, MMP-9 in the si-TM7SF4 group were lower than the si-NC group (P<0.001). However, the protein expression level of p21 in the si-TM7SF4 group was higher than the si-NC group (P<0.001). The inhibitory rate of T24 cells in the CAS+ pcDNA-TM7SF4 group was (21.45±2.46)%, lower than (64.06±4.49)% of the CAS+ pcDNA group (P<0.001), but the number of migration and invasion in the CAS+ pcDNA-TM7SF4 group were 75.66±6.57 and 59.35±5.40, higher than 40.43±3.85 and 30.25±3.32 in the CAS+ pcDNA group (P<0.001). The protein expression levels of TM7SF4, CyclinD1, MMP-2 and MMP-9 in the CAS+ pcDNA-TM7SF4 group were higher than the CAS+ pcDNA group (P<0.001), but the protein expression level of p21 was lower than the CAS+ pcDNA group (P<0.001). Conclusion: CAS may suppress the proliferation, migration and invasion of bladder cancer T24 cells by inhibiting the expression of TM7SF4.
Assuntos
Feminino , Humanos , Masculino , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Ciclina D1 , Flavonoides , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 9 da Matriz/genética , MicroRNAs/genética , RNA Mensageiro , Neoplasias da Bexiga Urinária/genéticaRESUMO
<p><b>OBJECTIVE</b>To summarize the experience of urethral reconstruction using circular fasciocutaneous flap for the treatment of complex anterior urethral strictures.</p><p><b>METHOD</b>The circular fasciocutaneous flap was harvested from the distal penile shaft or foreskin. Then the circular configuration was converted into longitudinal strip of skin which was rotated to ventral side to repair the exposured urethral strictures using the ventral onlay method. The surveillance protocol after urethroplasty was urination observation, regularly uroflowmetry and urethrography examination.</p><p><b>RESULTS</b>From Nov. 2006 to Oct. 2012, 15 cases were treated. The mean age was 45 years (20-76 years) and mean follow-up period was 18 months(3 months-3 years). Stricture was caused by chronic urethritis in 4 cases, long-term urethral catheterization in 3 cases, transurethral perfusion chem other aphy in 3 cases, transurethral prostatectomy in 3 cases and hypospadias after surgery in 2 patients. The mean stricture length was 7.0 cm (3.5-12.0 cm). The overall success rate was 80.0% (12/15). Recurrence stenosis was noted in 2 cases and diverticulum formation in 1 case.</p><p><b>CONCLUSIONS</b>The penile circular fasciocutaneous flap can be used for anterior urethral stricture in nearly any length. The flap has the characteristics of hairless, adequate mobile and length, well-vascularized pedicle and easy to harvest. The onlay reconstruction provides excellent cosmetic results, less trauma, higher success rate. Therefore it should be one of the preferred techniques for complex anterior urethral stricture repair.</p>
Assuntos
Humanos , Masculino , Seguimentos , Prepúcio do Pênis , Hipospadia , Cirurgia Geral , Pênis , Cirurgia Geral , Prostatectomia , Recidiva , Retalhos Cirúrgicos , Transplante , Uretra , Cirurgia Geral , Estreitamento Uretral , Patologia , Cirurgia Geral , Procedimentos Cirúrgicos Urológicos Masculinos , CicatrizaçãoRESUMO
<p><b>OBJECTIVE</b>To investigate the synergistical killing effect of docetaxel combined with ABT-737 on human prostate cancer cell line PC-3 by inducing apoptosis and further to determine the mechanism underlying such effect.</p><p><b>METHODS</b>PC-3 cells were treated with various concentrations of docetaxel or (and) ABT-737. Cell viability was determined using MTT assay. Apoptosis was assessed by fluorescence microscopy analysis of cells with condensed and segmented nuclei following staining with 4',6-diamidino-2-phenylindole (DAPI). Cellular DNA was stained with propidium iodide and flow cytometric analysis was performed to analyze the cell cycle distribution. Bcl-2, Bax, Bcl-xL and Mcl-1 protein changes were detected by Western blot. The activity of caspase-3 was measured using a colorimetric assay.</p><p><b>RESULTS</b>Docetaxel (20 nmol/L) combination with ABT-737 (400 nmol/L) for 48 hours, the cell viability was decreased to 19.7% ± 3.2% to compare with 44.2% ± 4.4% (t = 4.45) of docetaxel and 93.2% ± 1.8% of ABT-737 separately and there was a synergistic effect between the two drugs (CI = 0.8). Apoptosis rate of the combination group was higher than other two drugs. Docetaxel increased the cell number arrested in G(2)/M phase compared with control group (P < 0.05), but the combination treatment resulted in a significant arrest in the G(0)/G(1) phase. The combination treatment could significantly reduced the Bcl-2, Bcl-xL and Mcl-1 expression (F = 369.53, 57.89 and 32.77, all P < 0.05) and enhanced the activity of caspase-3 (419.7% ± 15.6%) (F = 207.33, P < 0.05).</p><p><b>CONCLUSIONS</b>The combination of ABT-737 with docetaxel can synergistically inhibit the proliferation of PC-3 cells through inducing apoptosis, which may be associated with cell cycle arrest, down-regulation of Bcl-2, Bcl-xL and Mcl-1 expression and activation of caspase-3.</p>
Assuntos
Humanos , Masculino , Apoptose , Compostos de Bifenilo , Farmacologia , Caspase 3 , Metabolismo , Ciclo Celular , Linhagem Celular Tumoral , Sinergismo Farmacológico , Proteína de Sequência 1 de Leucemia de Células Mieloides , Metabolismo , Nitrofenóis , Farmacologia , Piperazinas , Farmacologia , Neoplasias da Próstata , Metabolismo , Patologia , Proteínas Proto-Oncogênicas c-bcl-2 , Metabolismo , Sulfonamidas , Farmacologia , Taxoides , Farmacologia , Proteína bcl-X , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To improve the early diagnosis and therapeutic outcomes of testicular torsion.</p><p><b>METHODS</b>We retrospectively reviewed the clinical data of 49 cases of testicular torsion along with the results of their intratesticular color Doppler flow imaging (CDFI) and spermatic cord sonography.</p><p><b>RESULTS</b>Of the 49 cases, 42 showed abnormal intratesticular blood flow, including 3 cases of increased blood flow, while the other 7 presented no obvious abnormality. Morphological abnormality of the spermatic cord was found in 47 cases. Twenty-eight cases underwent testis removal, and the other 21 received detorsion and orchidopexy, in which 12 testes were preserved with normal size and blood flow.</p><p><b>CONCLUSION</b>Spermatic cord sonography and intratesticular CDFI play an important role in the early diagnosis of testicular torsion. And early surgical exploration contributes to the preservation of the testis.</p>
Assuntos
Adolescente , Adulto , Criança , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Diagnóstico Precoce , Orquidopexia , Estudos Retrospectivos , Cordão Espermático , Diagnóstico por Imagem , Torção do Cordão Espermático , Diagnóstico por Imagem , Cirurgia Geral , Ultrassonografia DopplerRESUMO
objective To investigate the effects of knocking down of miR-19a and miR-19b on the biological characteristics of SNB19 glioblastoma cells. Methods Oligonucleotides inhibitor of miR-19a and miR-19b (miR-19a inhibitor or miR-19b inhibitor) mediated by lipofectamine2000 were transfected to SNB19 cells to knock down miR-19a and miR-19b; control group (without transfection),group D (performing transfection with nonsense sequence) and group E (performing transfection with both miR-19a inhibitor and miR-19b inhibitor) were established. Real time PCR was conducted to detect the expressions ofmiR-19a and miR-19b in these groups after the transfection. The cell proliferation rate and cell cycle kinetics were detected by 3-(4, 5-Dime- -thylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and flow cytometry, respectively; the cell invasive ability was evaluated by Transwell assay.Results As compared with those in control group and group D, the expressions of miR-19a and miR-19b, proliferation activity and invasive ability of cells in the miR-19a/19b inhibitor transfected cells (group A/B) were significantly reduced (P<0.05). The expressions of miR-19a and miR-19b and the proliferation activity and invasive ability of cells 2, 3, 4 and 5 d after the transfection in group E were significantly reduced as compared with those in group A/B (P<0.05). Delayed cell cycle in group A/B and group E was noted as compared with that in control group and group D; and group E enjoyed more obviously delayed eell cycle than group A/B (P<0.05). Conclusion MiR-19a and miR-19b might be oncomiRs, and may be candidate target miRNAs for gene therapy of glioma.