PELO facilitates PLK1-induced the ubiquitination and degradation of Smad4 and promotes the progression of prostate cancer.

PLK1 and Smad4 are two important factors in prostate cancer initiation and progression. They have been reported to play the opposite role in Pten-deleted mice, one is an oncogene, the other is a tumor suppressor. Moreover, they could reversely regulate the PI3K/AKT/mTOR pathway and the activation of MYC. However, the connections between PLK1 and Smad4 have never been studied. Here, we showed that PLK1 could interact with Smad4 and promote the ubiquitination and degradation of Smad4 in PCa cells. PLK1 and PELO could bind to different domains of Smad4 and formed a protein complex. PELO facilitated the degradation of Smad4 through cooperating with PLK1, thereby resulting in proliferation and metastasis of prostate cancer cell. Changes in protein levels of Smad4 led to the alteration of biological function that caused by PLK1 in prostate cancer cells. Further studies showed that PELO upregulation was positively associated with high grade PCa and knockdown of PELO expression significantly decreased PCa cell proliferation and metastasis in vitro and vivo. PELO knockdown in PCa cells could enhance the tumor suppressive role of PLK1 inhibitor. In addition, blocking the interaction between PELO and Smad4 by using specific peptide could effectively inhibit PCa cell metastasis ability in vitro and vivo. Overall, these findings identified a novel regulatory relationship among PLK1, Smad4 and PELO, and provided a potential therapeutic strategy for advanced PCa therapy by co-targeting PLK1 and PELO.

Anticancer effect of miR-96 inhibitor in bladder cancer cell lines.

The present study aimed to investigate the role of microRNA-96 (miR-96) in the proliferation, invasion and apoptosis of bladder cancer cell lines, and the associated mechanisms. The expression of miR-96 and human ether-à-go-go-related (HERG1) potassium channel in the normal uroepithelium SV-HUC-1 cell line, and bladder cancer T24 and 5637 cell lines were examined using reverse transcription-polymerase chain reaction or/and western blotting. Transfection with miR-96 inhibitor or scrambled control (SC) was used to study the biological activities of miR-96 in bladder cancer cell lines. MTT, flow cytometric and Transwell assays were applied to detect cell viability, apoptosis and invasion, respectively. A dual-luciferase reporter assay was applied to determine the association between miR-96 and HERG1 expression. As demonstrated, miR-96 was highly expressed in the two bladder cancer cell lines, particularly in T24 cells. Following transfection with miR-96 inhibitor, miR-96 expression was significantly reduced in the T24 cell line, compared with SC. The miR-96 inhibitor suppressed cell proliferation and invasion, promoted apoptosis and arrested the cell cycle at the G1 phase. Consistently, HERG1 was also highly expressed in the two bladder cancer cell lines at the mRNA and protein level, but not in the normal uroepithelium cell line. The miR-96 inhibitor also significantly decreased HERG1 expression compared with SC. The results of the dual-luciferase reporter assay indicated that miR-96 directly targeted wild-type HERG1. In conclusion, miR-96 inhibitor exhibited anticancer effects on bladder cancer cells by inhibiting proliferation and invasion of cells, and promoting their apoptosis. HERG1 was an important target of miR-96. These results provided experimental evidence supporting miR-96 as a therapeutic target for patients with bladder cancer.

Experimental study of therapy of bone marrow mesenchymal stem cells or muscle-like cells/calcium alginate composite gel for the treatment of stress urinary incontinence.

OBJECTIVE: To explore the myoblast formation around the urethra and increase in urethral resistance of bone marrow mesenchymal stem cells or muscle-like cells/calcium alginate composite gel injection therapy and effect on LPP in SUI rat model. METHODS: Isolation, cultivation, and identification of SD rat bone marrow mesenchymal stem cell were performed. 5-Azacytidine was introduced to induce muscle-like cells. SUI was produced in 72 6-week-old female Sprague-Dawley rats, which were divided into four groups: stem cell-gel group, muscle-like cell-gel group, Gel group, and mock control group. One, 4, and 8 weeks after injection, the leak point pressure (LPP) was measured. HE staining of Desmin and α-skeletal muscle actin (α-SMA) were performed. RESULTS: At 4 and 8 weeks after injection in stem cell-gel group and muscle-like cell-gel group, growth of blood vessels gradually increased at gel edge, BMSC, and muscle-like cells gathered around the new blood vessels observed by fluorescence tracer, muscle-like cells grew into elongated spindle-like cells, Desmin, and α-SMA staining were obviously positive expression. LPP determinations of the mock control group compared with the Gel groups were significantly different. CONCLUSIONS: Compound of BMSC, muscle-like cells, and calcium alginate composite gel has the potential to differentiate into muscle cells in the microenvironment of SUI rat model. It is found by LPP measurement that the correlation between the increase in urethral resistance and the volume effect of calcium alginate gel is high.