ABSTRACT
Purpose@#To probe the effect and mechanism of androgen receptor antagonist MDV3100 on benign prostatic hyperplasia (BPH) of rats @*Methods@#BPH rat model was induced by testosterone propionate. Then antagomir-miR-21-3p or agomir-miR-21-3p was injected into rats before MDV3100 treatment. The prostate index was measured by weighing the wet weight of the rat prostate. The structural morphology of rat prostate was observed after hematoxylin & eosin staining. Immunohistochemistry was applied to evaluate the expression levels of Ki-6 and inflammatory cytokines (interleukin [IL]-6, IL-18, and tumor necrosis factor-α) in rat prostate tissues. Quantitative reverse transcription polymerase chain reaction was utilized for assessment of miR-21-3p expression, and Western blot for the performance of the phosphorylation levels of IKKα and p65. @*Results@#Injection of testosterone propionate caused increased prostate gland hyperplasia, heightened miR-21-3p level, and activated nuclear factor-kappa B (NF-κB) signaling pathway. Additionally, BPH was accompanied by inflammatory response, as evidenced by enhanced expressions of Ki-67 and inflammatory cytokines. MDV3100 exposure ameliorated BPH and suppressed miR-21-3p expression. Overexpression of miR-21-3p intensified BPH and inflammation level, while knockdown of miR-21-3p relieved BPH. The coeffect of miR-21-3p upregulation and MDV3100 subjection led to higher inflammatory response, elevated phosphorylation levels of IKKα and p65 than MDV3100 treatment alone. @*Conclusions@#Androgen receptor antagonist MDV3100 alleviates BPH and inflammatory response through miR-21-3p downregulation and NF-κB signaling pathway blockade.
ABSTRACT
Purpose@#To probe the effect and mechanism of androgen receptor antagonist MDV3100 on benign prostatic hyperplasia (BPH) of rats @*Methods@#BPH rat model was induced by testosterone propionate. Then antagomir-miR-21-3p or agomir-miR-21-3p was injected into rats before MDV3100 treatment. The prostate index was measured by weighing the wet weight of the rat prostate. The structural morphology of rat prostate was observed after hematoxylin & eosin staining. Immunohistochemistry was applied to evaluate the expression levels of Ki-6 and inflammatory cytokines (interleukin [IL]-6, IL-18, and tumor necrosis factor-α) in rat prostate tissues. Quantitative reverse transcription polymerase chain reaction was utilized for assessment of miR-21-3p expression, and Western blot for the performance of the phosphorylation levels of IKKα and p65. @*Results@#Injection of testosterone propionate caused increased prostate gland hyperplasia, heightened miR-21-3p level, and activated nuclear factor-kappa B (NF-κB) signaling pathway. Additionally, BPH was accompanied by inflammatory response, as evidenced by enhanced expressions of Ki-67 and inflammatory cytokines. MDV3100 exposure ameliorated BPH and suppressed miR-21-3p expression. Overexpression of miR-21-3p intensified BPH and inflammation level, while knockdown of miR-21-3p relieved BPH. The coeffect of miR-21-3p upregulation and MDV3100 subjection led to higher inflammatory response, elevated phosphorylation levels of IKKα and p65 than MDV3100 treatment alone. @*Conclusions@#Androgen receptor antagonist MDV3100 alleviates BPH and inflammatory response through miR-21-3p downregulation and NF-κB signaling pathway blockade.
ABSTRACT
Objective To investigate the effect of silencing Notch1 gene by RNA interference on the proliferation,apoptosis and Akt/mTOR signaling pathway in clear renal cell carcinoma.Methods The optimal segment targeting Notch1 gene was designed and transfected into 786-O cells by Lipofectamine TM2000.The Notch1 mRNA and protein were detected by RT-PCR and Western blot.The proliferation rate of 786-O cells was evaluated by MTT and the variation of apoptosis was measured by TUNEL.The protein expression level of apoptosis-related protein Bcl-2,caspase-3,caspase-9,and signaling pathway protein Akt,p-Akt,p-mTOR,p-P70S6K were detected by Western blott.Results Notchl mRNA and protein was markedly suppressed by the siRNA targeting Notch1.Treated with 0,40,60,80,100 and 120 nmol/L of Notch1 siRNA for 24 hours,cell proliferation rates were (98.51 ± 1.33) ％,(87.34 ± 2.26) ％,(64.72 ± 3.24)％,(57.68 ±3.32)％,(31.91 ± 1.85)％ and (19.27 ±2.73)％,and the difference was statistically significant (P ＜ 0.01).Treated with 0,40,80,and 120 nmol/L of Notchl siRNA for 24 hours,apoptosis rates were (7.6 ± 3.8) ％,(21.5 ± 4.8) ％,(32.3 ± 3.5) ％,and (46.3 ± 4.7％),the difference was statistically significant (P ＜ 0.05).Decreased expression of Akt signaling pathway proteins p-Akt,p-mTOR,p-70S6K and apoptosis-related protein Bcl-2,procaspase-3 was detected,but no change in the total protein of Akt.Conclusions Depletion of Notch1 gene could inhibit cell growth and induce apoptosis in 786-O cell line.It inhibits Akt/mTOR signaling pathway by dephosphorylated.
ABSTRACT
<p><b>BACKGROUND</b>Currently, cystoscopy and urine cytology are standard modalities in therapy monitoring and follow-up of bladder carcinoma (BC). Cystoscopy is an invasive and uncomfortable procedure while cytology has a limited value because it is operator-dependent and has low sensitivity. This study was to assess the accuracy of ImmunoCyt in detecting BC by comparing it with cytology using systemic analyses of studies published in both English and Chinese.</p><p><b>METHODS</b>Cochrane systematic evaluation was used to search through MEDLINE, EMBASE, Cochrane Library, CMCC, and CNKI for studies regarding ImmunoCyt and cytology for detection of BC. Data were extracted and analyzed by the software MetaDiSc 1.4.</p><p><b>RESULTS</b>In total 42 relevant studies were searched, of which 15 were enrolled and 12 491 patients were included. Heterogeneity, except for threshold effects, was found within these studies. A meta-analysis was performed using the random effect model. Pooled accuracy indicators like sensitivity, specificity, and diagnostic odds ratio of ImmunoCyt™ and cytology were 0.75 (0.73-0.77) vs. 0.45 (0.43-0.48), 0.73 (0.72-0.74) vs. 0.97 (0.96-0.97), and 10.97 (7.53-15.99) vs. 16.40 (10.57-25.46), respectively. The sensitivity of both was increased with the increase of tumor grade and stage. The area under summary receiver operating characteristics curve was 0.834 4 and 0.853 4 and the Q index 0.766 7 and 0.785 3 for ImmunoCyt and cytology, respectively. Combination of both can obviously improve the accuracy of diagnosis.</p><p><b>CONCLUSIONS</b>ImmunoCyt has a high sensitivity in detecting BC, but its specificity is low. As an important adjunct, ImmunoCyt™ can not replace cytology, but combined with cytology it could improve sensitivity with high specificity in the detection and postoperative monitoring of BC.</p>
Subject(s)
Humans , Immunohistochemistry , Sensitivity and Specificity , Urinary Bladder Neoplasms , DiagnosisABSTRACT
Objective To investigate phenyhexyle isothiocyanate (PHI) modulating histone acetylation and inhibiting Akt signaling pathway in prostate cancer cell line PC3 in vitro. Methods Apoptotic cells were measured by TUNEL assay. Histone acetylated H3, H4 and the Akt protein signaling pathway (Akt, p-Akt, mTOR, p-mTOR, p70S6K and p-p70S6K) were detected by Western blot. Results Apoptotic cells increased after exposure to PHI with concentration dependent. PHI significantly induced an accumulation of histone acetylated H3, H4. The change of Akt, mTOR, p70S6K proteins was not observed. Phosphorylation of Akt (p- Akt), mTOR (p-mTOR) and p70S6K (p-p70S6K) decreased after exposure to PHI for 7 h. Conclusions PHI can induce histone acetylation H3, H4 accumulation. PHI inhibits Akt signaling pathway resulting cell apoptosis. It might be a new anticancer agent.