ABSTRACT
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.
Subject(s)
Enhancer of Zeste Homolog 2 Protein/genetics , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , Prostatic Neoplasms/genetics , Prostatic Neoplasms/pathology , RNA Interference , 3' Untranslated Regions , Aged , Aged, 80 and over , Apoptosis , Biomarkers, Tumor , Cell Line, Tumor , Cell Proliferation , Enhancer of Zeste Homolog 2 Protein/metabolism , Epithelial-Mesenchymal Transition/genetics , Humans , Male , Middle Aged , Neoplasm Grading , Neoplasm Staging , Prostatic Neoplasms/metabolismABSTRACT
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.
Subject(s)
Apoptosis/drug effects , Autophagy/drug effects , Hydroxychloroquine/pharmacology , Prostate/cytology , Animals , Male , Orchiectomy , Random Allocation , Rats , Rats, Sprague-DawleyABSTRACT
OBJECTIVE: To investigate the effects of LY294002, a phosphatidylinositol 3-kinase inhibition, on sperm motility in asthenozoospermia patients in vitro, and further analyze the possible molecular mechanism. METHODS: Sperm aseptically obtained by masturbation and prepared by swim-up technique from 10 patients with asthenozoospermia and 10 healthy fertile men were incubated with different concentrations of LY294002. Measurements of motility were carried out at 10, 30 and 60 min in all specimens by CASA. RESULTS: The sperm in asthenozoospermia patients treated with LY294002 showed a significant increase in sperm progressive motility, the percentage of motile cells, VSL and VAP. CONCLUSION: LY294002 can enhance the motility of sperm in asthenozoospermia patients in vitro.
