ARPC4 promotes bladder cancer cell invasion and is associated with lymph node metastasis.

The significance of actin-related protein 2/3 complex subunit 4 (ARPC4) expression in bladder cancer, and its potential role in the invasion and migration of bladder cancer cells, has yet to be determined. This study was to identify the correlation between ARPC4 and lymph node metastasis, and to determine the role of ARPC4 in the invasive migration of T24 bladder cancer cells. One hundred and ninety-eight bladder cancer tissues and 40 normal bladder and lymph node tissues were examined. Tissue microarrays were constructed and subjected to immunohistochemical stating for ARPC4. Multiple logistic analysis was used to determine risk factors associated with bladder cancer metastasis. ARPC4 expression in T24 bladder cancer cells was suppressed using small interfering RNA and changes in protein levels were determined by Western blot analysis. The proliferation of bladder cancer cells after knocking down of ARPC4 was determined by cell counting kit-8. The effects of ARPC4 knockdown on T24 cell invasion and migration was determined using transwell and wound healing assays. Immunofluorescence analysis was performed to examine changes in pseudopodia formation and actin cytoskeleton structure. The expression of ARPC4 was elevated in bladder cancer tissues than normal tissues (84.3% vs 27.5%, P < 0.001). The multivariate logistic analysis demonstrated that the level of ARPC4, as a risk factor, was correlated with lymphatic metastasis (P < 0.05). ARPC4 knockdown attenuated proliferation, migration, invasion, and pseudopodia formation in T24 cells. ARPC4 expression, as a risk factor, is associated with lymphatic metastasis and is upregulated in bladder cancer tissues in comparison with normal tissues. Inhibition of ARPC4 expression significantly attenuates the proliferation, migration, and invasion of bladder cancer cell, possibly due to defects in pseudopodia formation.

Upregulation of Arp2 expression is associated with the prognosis and prediction of lymph node metastasis in bladder urothelial carcinoma.

OBJECTIVE: Upregulation of actin-related protein 2/3 complex subunit 2 (Arp2) is observed in various tumors, but its expression pattern has not been revealed in bladder urothelial carcinoma (BUC). The purpose of this study was to investigate the role of Arp2 expression in the pathological features and the prognosis as well as lymph node metastasis of BUC. METHODS: A total of 228 tissue specimens from BUC patients who underwent a radical cystectomy were collected. In addition, 40 lymph node specimens and 40 normal bladder tissue specimens as controls were used. All of the specimens were used to construct a tissue microarray and were subsequently subjected to immunohistochemical staining for Arp2 expression. Logistic and Cox regression analyses and Kaplan-Meier curve analysis were applied to analyze the relation between Arp2 expression and multiple clinical features of patients with BUC. RESULTS: Immunohistochemical staining observation showed that Arp2 was mainly expressed in the cytoplasm and nucleus of positive cells and that Arp2 expression in BUC was significantly higher than that in normal bladder tissues. Arp2 expression in BUC tissues was associated with tumor size, tumor multiplicity, tumor stage, tumor grade, and lymph node metastasis (P < 0.05). Logistic regression analysis demonstrated that Arp2 expression was an independent risk factor for lymph node metastasis of BUC (P < 0.05). Kaplan-Meier curve analysis showed that increased Arp2 expression was associated with a shortened recurrence-free survival (RFS) and overall survival (OS) of BUC patients after radical cystectomy (P < 0.05). CONCLUSION: These findings suggest that Arp2 is significantly upregulated in BUC tissues when compared with normal bladder tissues, and that Arp2 expression is an independent predictor for lymph node metastasis, RFS, and OS.

Fasudil inhibits proliferation and collagen synthesis and induces apoptosis of human fibroblasts derived from urethral scar via the Rho/ROCK signaling pathway.

Fasudil has shown antifibrotic effects in various fibrotic diseases. However, its effects on human urethral fibroblasts are unknown. This study evaluated the effects of fasudil on cellular proliferation, migration, apoptosis, and collagen synthesis in human fibroblasts derived from urethral scar tissues. Human urethral scar fibroblasts were cultured by explant and incubated for 24 h or 48 h with fasudil (12.5, 25, 50 µmol/L) with or without transforming growth factor ß1 (TGF-ß1, 10 ng/mL), or left untreated (control). Cell proliferation and migration was determined by MTT assay and Transwell chambers, respectively. Apoptosis was measured by flow cytometry. Levels of α-smooth muscle actin (α-SMA), myosin light-chain phosphatase (MLCP), LIM domain kinase 1 (LIMK1), phospho-cofilin (p-cofilin), collagen I, and collagen III were determined by Western blot. Compared with the control group, TGF-ß1 was associated with a significant increase in urethral fibroblast proliferation and migration, and α-SMA, MLCP, LIMK1, p-cofilin, collagen I, and collagen III levels. Compared with the control group, fasudil (with or without TGF-ß1), significantly and negatively correlated, in a dose-dependent manner, with the proliferation and migration of urethral fibroblasts, as well as α-SMA, MLCP, LIMK1, p-cofilin, collagen I, and collagen III levels. Moreover, fasudil significantly induced apoptosis of fibroblasts induced by TGF-ß1. Higher concentrations of fasudil (50 µmol/L) were associated with greater cell apoptosis without TGF-ß1 stimulation compared with the normal control group. Fasudil, with or without TGF-ß1 stimulation, may inhibit human urethral fibroblasts proliferation, migration, apoptosis, and collagen synthesis via the Rho/ROCK signaling pathway.

[Risk factors of ISUP Modified Gleason score upgrading after radical prostatectomy].

OBJECTIVE: To investigate the factors upgrading the International Society of Urological Pathology (ISUP) Gleason score using the specimens from preoperative prostatic biopsy and radical prostatectomy. METHODS: A total of 164 patients diagnosed with prostate cancer by biopsy underwent radical prostatectomy. We retrospectively analyzed their age, prostate volume, preoperative PSA level, PSA density (PSAD) , the time interval between biopsy and surgery, the number of positive punctures, positive surgical margin, seminal vesicle invasion, lymphatic invasion, and Gleason scores from biopsy and prostatectomy. We also determined the predictors of Gleason score upgrading by logistic regression analysis. RESULTS: Of the 164 cases analyzed, 95 (57.93% ) showed a consistency between the Gleason score of preoperative prostatic biopsy and that after radical prostatectomy, 55 (33.54% ) increased and 14 (8.52%) decreased after prostatectomy as compared with preoperative biopsy. The prostate volume (P < 0.01) and biopsy score (P < 0.05) were independent predictors of Gleason score upgrading. The risk of Gleason score upgrading was 27 times higher in the patients with the prostate volume ≤ 25 ml and 9 times higher in the 25-40 ml group than in the > 60 ml group (P < 0.05). CONCLUSION: Low Gleason score of biopsy (≤ 6) and small prostate volume (≤ 40 ml) may be the predictors of Gleason score upgrading after radical prostatectomy.