Hydroxychloroquine Blocks Autophagy and Promotes Apoptosis of the Prostate after Castration in Rats.

Autophagy is an important pro-survival mechanism and closely related to apoptosis. The aim of this study was to investigate whether hydroxychloroquine (HCQ) blocks autophagy and promotes apoptosis of the prostate after castration. METHODS: Thirty-six male SD rats were randomly divided into 3 groups (n = 12): control group (sham operation), castration group, and HCQ group (castrated and treated with HCQ). On day 7, all mice were executed and prostates were isolated. The morphological changes of prostates were observed by light microscope, and the ultrastructure changes were observed under scanning electron microscope (SEM). The protein expression of Beclin-l, P62, caspase-3, Bcl-2, and Bax was assessed by immunohistochemical analyses. The mRNA expression of microtubule-associated protein light chain 3 (LC3) and autophagy-related gene 5 (Atg5) was detected by RT-PCR. RESULTS: Prostates of castration group shrank remarkably and prostates of HCQ group shrank more remarkably than castration group. Cytolysosomes were visible in the prostates of the castration group under SEM. Immunohistochemistry showed that the protein of Beclin-1 increased in the castration group compared to the control group, while decreased in the HCQ group compared to the castration group. While P62 protein moderately dyed in the control group and weakly dyed in the castration group, it strongly dyed in the HCQ group. Caspase-3 and Bax protein were weakly dyed in the control group but moderately dyed in the castration group and strongly dyed in the HCQ group. The expressions of apoptosis suppressor Bcl-2 were reduced in the castration group and further reduced in the HCQ group compared to the castration group. RT-PCR revealed that the mRNA of LC3 and Atg5 in the castration group increased compared to the control group, while decreased after treated with HCQ. CONCLUSION: Autophagy increased after castrated in prostates, while decreased after treated with HCQ; all these indicated that HCQ blocked autophagy and then promoted prostate apoptosis of castrated mice.

MicroRNA-144 Suppresses Prostate Cancer Growth and Metastasis by Targeting EZH2.

BACKGROUND: Prostate cancer is a common malignant tumor with a high incidence. MicroRNAs (miRNAs) have been shown to be important post-transcriptional regulators during tumorigenesis. This study aimed to explore the effect of miR-144 on PCa proliferation and apoptosis. MATERIAL AND METHODS: The expression of miR-144 and EZH2 were examined in clinical PCa tissues. PCa cell line LNCAP and DU-145 was employed and transfected with miR-144 mimics or inhibitors. The correlation between miR-144 and EZH2 was verified by luciferase reporter assay. Cell viability, apoptosis and migratory capacity were detected by CCK-8, flow cytometry assay and wound healing assay. The protein level of EZH2, E-Cadherin, N-Cadherin and vimentin were analyzed by western blotting. RESULTS: miR-144 was found to be negatively correlated to the expression of EZH2 in PCa tissues. Further studies identified EZH2 as a direct target of miR-144. Moreover, overexpression of miR-144 downregulated expression of EZH2, reduced cell viability and promoted cell apoptosis, while knockdown of miR-144 led to an inverse result. miR-144 also suppressed epithelial-mesenchymal transition level of PCa cells. CONCLUSION: Our study indicated that miR-144 negatively regulate the expression of EZH2 in clinical specimens and in vitro. miR-144 can inhibit cell proliferation and induce cell apoptosis in PCa cells. Therefore, miR-144 has the potential to be used as a biomarker for predicting the progression of PCa.

[Feasibility of muscle-derived cell autotransplantation as a treatment for post-prostatectomy urinary incontinence].

OBJECTIVE: To explore the feasibility of muscle-derived cell autotransplantation in the treatment of post-prostatectomy urinary incontinence. METHODS: Skeletal muscle-derived cells (MDC) were isolated and purified by replate technique from 6 female SD rats, and then transduced with adenovirus carrying Lac-Z gene. About 5 x 10(6) of the transduced cells were injected autologously into the bladder neck of the animals. Tissues were harvested after 5 and 15 days for histological examination and X-gal staining. RESULTS: At 5 and 15 days after the autologous MDC transplantation, histological examination revealed no apparent sign of inflammation and inflammatory cell invasion, and X-gal staining showed a large number of cells dyed blue, indicating the survival of the autologous cells. CONCLUSION: Autotransplanted MDCs can survive permanently. Autologous muscle stem cell injection can be an effective treatment for post-prostatectomy urinary incontinence.

[Experimental study of contralateral testicular changes after unilateral testicular torsion in rats].

OBJECTIVE: To study the changes of the contralateral testicular histology and germ cell apoptosis after unilateral testicular torsion (UTT) and to determine whether the contralateral testis is injured or not. METHODS: Sixty SD male rats were divided into control group (12 rats) and experimental group(48 rats). The former underwent sham operation of the left testis under general anaesthesia. The latter underwent left testis torsion(720 degrees) for 6 h, and then 4 of them were sacrificed and the other 44 were subdivided into the torsed testis untwisted group (22 rats) and the torsed testis removal group (22 rats), 7-8 rats were sacrificed and both testes (twisted and untwisted) were removed 1 day, 1 week and 4 weeks after surgery. All testes underwent histological and germ cell apoptosis examination. RESULTS: There were significant histological changes in the contralateral testis, and the germ cell apoptosis was increased greatly in the contralateral testis. CONCLUSIONS: UTT can cause contralateral testicular injury, whose mechanism may be related to reperfusion, and torsed testis removal can prevent or reduce damage to the contralateral testis.