Translational Andrology and Urology
·
2024
·Vol. 13
·期 9
·2055
·
IF 1.9
Shoubin Li,Chunhong Yu,Helong Xiao,Qingle Xu,Bo Gao,Liuxiong Guo,Zhanxin Sun,Junjiang Liu
Background Benign prostatic hyperplasia (BPH) is a common disease among older men characterized by non-malignant proliferation of epithelial cells and inflammation. Nitric oxide synthase traffic inducer (NOSTRIN) is a pleiotropic regulator of endothelial cell function and signaling and exerts anti-inflammatory, anti-proliferation, and modulating nuclear factor-kappa B (NF-κB) signaling effects. Its expression and function in BPH tissues and prostate epithelial cells are unknown. The study aims to investigate the expression and functions of NOSTRIN in BPH, and its possible molecular mechanism. Methods The BPH model was constructed in male Institute of Cancer Research (ICR) mice using 5 mg/kg/day testosterone propionate (TP) for 30 days, and the model was evaluated by detecting prostate index, prostate epithelial thickness, and prostate-specific antigen (PSA) expression. Dihydrotestosterone (DHT, 10 nM)-induced in vitro model of human prostate epithelial cells (RWPE-1) was established. We generated lentivirus-harboring human NOSTRIN . The mRNA expression was detected by real-time quantitative polymerase chain reaction (PCR) assay; the protein expression or localization was detected by western blot assay, immunohistochemistry, or immunofluorescence staining. Cell proliferation was assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and 5-ethynyl-2'-deoxyuridine (EdU) staining. Reactive oxygen species (ROS) production was observed by dihydroethidium staining. Nitric oxide (NO) and malondialdehyde (MDA) levels and superoxide dismutase (SOD) activity were detected using commercial kits. Enzyme-linked immunosorbent assay (ELISA) was used to determine levels of interleukin 1 beta (IL1B), interleukin 6 (IL6), interferon gamma (IFNG), and tumor necrosis factor (TNF). Results NOSTRIN expression was significantly inhibited in the TP-induced ICR mouse BPH model and DHT-induced model of RWPE-1 proliferation. Protein expression of the BPH-related and proliferation markers PSA and proliferating cell nuclear antigen (PCNA) was suppressed in NOSTRIN-overexpressing RWPE-1 cells exposed to DHT. NOSTRIN overexpression notably inhibited the RWPE-1 cell proliferation in vitro , as evidenced by MTT and EdU staining. NOSTRIN overexpression significantly decreased the expression of cell cycle-related proteins cyclin dependent kinase 4 (CDK4) and cyclin D1 (CCND1) in vitro . The production of ROS, NO, and lipid peroxidation products MDA was inhibited by NOSTRIN overexpression in vitro , while the SOD activity was increased. NOSTRIN overexpression reduced the mRNA expression of inflammatory mediator nitric oxide synthase 2 (NOS2) and inhibited the mRNA expression and secretion of pro-inflammatory cytokines IL1B, IL6, IFNG, and TNF in vitro . The mechanistic studies revealed an increased phosphorylation of NF-κB p65 in vivo and in vitro . Remarkably, NOSTRIN overexpression notably inhibited the protein expression of phospho-NF-κB p65 in vitro . Conclusions NOSTRIN is involved in BPH by inhibiting proliferation, oxidative stress, and inflammation in prostate epithelial cells. These functions may act through the inhibition of NF-κB signaling.
研究领域:
泌尿学
·
细胞生物学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Oncology Letters
·
2020
·Vol. 20
·期 4
·1-1
·
IF 2.311
Peng He,Guoguang Xiong,Wei Guo,Guanjun Jiang,Yun Li,Hao Li
Long non‑coding RNA colon cancer associated transcript 2 (CCAT2) is dysregulated in a number of different types of human cancer, and affects cancer progression via the Wnt/β‑catenin signaling pathway. However, the roles of CCAT2 and the Wnt/β‑catenin signaling pathway in prostate cancer (PCa) are not completely understood. The present study aimed to investigate the potential mechanism of CCAT2 in PCa. In the present study, the reverse transcription‑quantitative PCR (RT‑qPCR) results indicated that CCAT2 expression was significantly upregulated in PCa tissues, and DU145 and PC3 cell lines compared with normal prostate tissues and the epithelial RWPE‑1 cell line, respectively. Functional assays indicated that CCAT2 downregulation inhibited DU145 and PC3 cell proliferation, cell cycle, migration and invasion. In addition, the luciferase reporter assay, RT‑qPCR and western blotting results indicated that CCAT2 regulated transcription factor 7 like 2 (TCF7L2) expression by binding to microRNA‑217. Further western blotting and TOPFlash assays indicated that CCAT2‑knockdown inhibited the Wnt/β‑catenin signaling pathway in DU145 and PC3 cell lines by inhibiting the expression of TCF7L2. However, CCAT2‑knockdown‑mediated effects were reversed by the Wnt/β‑catenin signaling pathway activator lithium chloride (LiCl). Further cell experiments suggested that LiCl treatment reversed CCAT2‑knockdown‑mediated inhibition of PCa cell proliferation, cell cycle, epithelial‑mesenchymal transition, migration and invasion. Overall, the results indicated that CCAT2 regulated PCa via the Wnt/β‑catenin signaling pathway; therefore, CCAT2 may exhibit key role during the progression of PCa and may serve as a therapeutic target for the disease.
研究领域:
肿瘤学
·
分子生物学
·
细胞生物学
·
癌症遗传学
✓ 已验证
DOI
PubMed
高影响因子
RSC Advances
·
2016
·Vol. 6
·期 92
·89484-89491
·
IF 3.289
Yingshuai Liu,Jin Xie,Zeying Zhang,Zhisong Lu
O-GlcNAcylation is a very important post-translational modification of nucleocytoplasmic proteins. Aberrant O-GlcNacylation has been found in a variety of human diseases including many types of cancers, diabetes, and Alzheimer's disease. However, analysis of protein O-GlcNacylation still remains a great challenge due to its low levels and labile nature. Herein, a highly sensitive and reliable colorimetric method is developed by use of wheat germ agglutinin (WGA) lectin for specific recognition of O-GlcNAc and gold nanoparticle (AuNP)-catalyzed copper deposition for signal amplification. In presence of the target molecule, AuNP is specifically introduced onto the sensing surface and then catalyzes the copper deposition. An Au@Cu core–shell structure is produced from the deposition. The obtained copper shell on AuNP is dissolved by FeCl3 due to the oxidation–reduction reaction between copper and Fe3+, resulting in Cu2+ and Fe2+, which subsequently form a red coordination complex in the presence of bathophenanthrolinedisulfonic acid disodium salt (BPT2−). Thus, the amount of the red complex is proportional to the level of O-GlcNAc. Qualitative O-GlcNAc detection is realized by direct readout with the naked eye, while the quantitative analysis is achieved by measurement of the absorbance using a UV-Vis spectrophotometer. With increasing time, more and more copper is deposited on the surface of AuNP, thus achieving significant signal amplification. Under optimal conditions, an ultralow limit of detection (LOD) down to 0.02 pg mL−1 is achieved with GlcNAc-conjugated BSA as a model analyte. The developed platform was also successfully applied to analyze O-GlcNAcylation levels in prostate cancer cell line PC-3 and normal cell line RWPE-1. The results obtained by the proposed method are in good agreement with the data from protein microarray assays, demonstrating its great potential for sensitive and reliable O-GlcNAcaylation detection in fundamental research and in cancer diagnosis.
研究领域:
生物化学
·
分析化学
·
癌症生物学
·
蛋白质科学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）