BACKGROUND AND OBJECTIVE: Clusterin promotes cell proliferation, motility and invasiveness in human renal cell carcinoma (RCC) cells but the underlying molecular mechanisms of this action are largely unknown. The aim of this study was to investigate the effects of clusterin on cancer cell growth, i nvasion and S100A4 expression and to determine the effects of clusterin on in vitro cell proliferation and migration and in vivo tumour growth in RCC cells. METHODS: We have established stable transfectants of highly invasive Caki-1 human RCC cells with expression of clusterin shRNA targeting clusterin (Caki-1/clusterin shRNA). We also established stable transfectants of 786-O human RCC cells with expression of clusterin cDNA plaismid (786-O/clusterin cDNA). Clusterin and S100A4 expression was detected by reverse transcription (RT) PCR and western blot assay; Caki-1/clusterin shRNA and 786-O/clusterin cDNA clones were subjected to in vitro-invasion assays. Cell viability and cell growth was assessed in MTT and clonogenic assay. Specific small interfering RNA was employed to down-regulate S100A4. The expression plasmid for S100A4 (pCMV-S100A4) was used to upregulate S100A4. Caki-1/clusterin shRNA clones were injected subcutaneously in nude mice to determine tumour growth and cancer cell invasiveness in vivo. Xenograft tumour tissues were assessed by immunohistochemistry and frozen tissues were used for the detection of S100A4 and clusterin. RESULTS: Overexpression of clusterin increased cell invasiveness; and targeting clusterin reduced cell invasiveness in vitro. This increase in cell invasiveness was mediated by S100A4. Targeting clusterin decreased cell proliferation and down-regulated cellular S100A4 levels in Caki-1 cells; Overexpression of clusterin increased cell proliferation and up-regulated cellular S100A4 levels in 786-O cells; Stable Caki-1/clusterin shRNA transfectants produced smaller xenograft tumours containing reduced S100A4 protein levels in vivo. Stable 786-O/clusterin cDNA transfectants produced larger xenograft tumours containing increased S100A4 protein levels in vivo. CONCLUSION: Our results indicate that clusterin promotes growth and invasion in RCC cells in vitro and in vivo through upregulation of S100A4; And targeting clusterin confers growth inhibitory and anti-invasive properties in RCC cells in vitro and in vivo through a down-regulation of S100A4. These findings provide the rationale for future oncostatic strategies aimed at suppressing clusterin-mediated signal transduction pathways as a novel therapeutic approach in human RCC.
SC79, an AKT activator, has been reported to protect experimental ischemia-elicited neuronal death, brain injury, and myocardiocyte hypoxia/reoxygenation (H/R) injury. However, the protection of SC79 from renal ischemia-reperfusion (I/R) injury and the precise mechanisms involved are unknown. Here, we investigated the effects of SC79 in renal tubular epithelial cells in vitro and in mouse kidney in vivo following hypoxia-reoxygenation (H/R) and renal I/R injury.The kidneys of Sprague-Dawley rats were subjected to 30 min of warm ischemia followed by 24 h of reperfusion. Murine renal tubular epithelial NRK-52E cells were subjected to hypoxia for 6 h and reoxygenation for 24 h. The NRK-52E cells and the renal I/R injury model were treated with SC79 and/or LY294002 at different times and concentrations. Serum creatinine (Cr) concentration, renal histology, cellular viability, and cell apoptosis were assessed. Levels of phospho-Akt, bad, Bim, bax, bcl-2, and bcl-XL in NRK-52E cells and renal tissues were determined by western blotting.SC79 improved viability and inhibited apoptosis in NRK-52E cells following H/R. SC79 decreased serum Cr and markedly improved pathology and decreased cell apoptosis in kidneys following I/R. In addition, SC79 promoted the expression of phospho-Akt, bcl-2, and bcl-XL, and decreased the expression levels of bid, bax, and bim. PI3K inhibitor (LY294002) pre-treatment completely abolished these effects of SC79.The protective role of SC79 against H/R of NRK-52E cells or renal I/R injury is related to activation of phosphorylation of AKT, resulting in a decrease in the pro-apoptotic proteins bim, bax, and bad and an increase in the anti-apoptotic proteins bcl-2 and bcl-XL induced by cell H/R and renal I/R injury.
MicroRNAs (miRNAs) are involved in cancer development and progression. Renal cell carcinoma (RCC) frequently undergoes metastasis and has a high mortality rate. The current study measured miRNA‑126 (miR‑126) expression levels in 128 pairs of clear cell RCC and adjacent normal kidney tissue samples by reverse transcription‑quantitative polymerase chain reaction, and analyzed the association between miR‑126 and various clinicopathological parameters. In addition, cell proliferation, wound healing and cell invasion assays were conducted using RCC cells overexpressing miR‑126. Potential miR‑126 target genes and the signaling pathways that may be regulated by miR‑126 were then examined. miR‑126 expression was significantly reduced in patients with metastatic RCC compared with patients without metastasis. Consistently, overexpression of miR‑126 in RCC cells significantly inhibited cell proliferation, migration and invasion in vitro compared with negative control miRNA. A luciferase reporter assay demonstrated that miR‑126 targets Rho associated coiled‑coil containing protein kinase 1 (ROCK1) by directly binding the 3'‑untranslated region. Furthermore, western blotting identified miR‑126 as an important regulator of the AKT and extracellular signal‑regulated 1/2 signaling pathways. The results of the present study indicate that miR‑126 inhibits RCC cell proliferation, migration and invasion by downregulating ROCK1. These findings suggest that miR‑126 may be valuable as a potential target for therapeutic intervention in RCC.
The global incidence of metabolic syndrome (MetS) is dramatically increasing. Considerable interest has been devoted to the relationship between MetS and prostate cancer (PCa) risk. However, few studies have examined the association between MetS and PCa progression. This retrospective study consisted of 1016 patients with PCa who received radical prostatectomy. The association between MetS and pathological features was evaluated using logistic regression analysis. Compared with patients without MetS, those with MetS indicated an increased risk of prostatectomy Gleason score (GS) ≥8 (odds ratio [OR] =1.670, 95% confidence interval (CI) 1.096–2.545, P= 0.017), and a 1.5-fold increased risk of pT3–4 disease (OR = 1.583, 95% CI 1.106–2.266, P= 0.012). The presence of MetS was an independent predictor of lymph node involvement (OR = 1.751, 95% CI 1.038–2.955, P= 0.036). Furthermore, as the number of MetS components accumulated, the risk of a GS ≥ 8 increased. The present study indicates a significant association between MetS and advanced PCa. The results need to be evaluated in large-scale prospective cohorts.
Purpose: The aim of this study was to investigate the relationship between single nucleotide polymorphisms (SNPs) within the 3′ untranslated region of the microRNA (miRNA)-binding site of the ADAM metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS5) gene and the risk of knee osteoarthritis (KOA) and its mechanism. Materials and Methods: Sanger sequencing was used to determine the genotypes of three ADAMTS5 gene SNPs (rs3171407, rs229071, and rs229077) from 310 patients suffering from osteoarthritis of the knee joint (KOA) and 310 healthy controls. The levels of the miRNAs, thsa-miR-103b, hsa-miR-144-3p, and hsa-miR-105-5p in plasma, and the level of ADAMTS5 mRNA in the articular cartilage of 132 patients with KOA were detected by real-time quantitative PCR. Results: The G allele at the rs3171407 locus of the ADAMTS5 gene was demonstrated to be a protective genetic factor for KOA. In addition, the risk of developing KOA was significantly increased in subjects carrying the C allele at the rs229071 locus. The risk of developing KOA in carriers of the G allele at locus rs229077 was 1.49 times greater than those with the A allele. The level of the hsa-miR-144-3p was increased, and the expression level of the ADAMTS5 protein was decreased in carriers of the T allele at the rs229071 locus. In carriers of rs229077 locus A allele the hsa-miR-105-5p level was increased and the expression level of ADAMTS5 protein was decreased. Conclusion: SNPs at the rs3171407, rs229071, and rs229077 loci of the ADAMTS5 gene are related to the risk of OA. The likely mechanism underlying these observations is that these SNPs affect the regulation of ADAMTS5 protein expression through miRNAs; however, this needs to be verified using in vivo models.
To explore the effect of anti-P-selectin monoclonal antibody (mAb) on renal ischemia/reperfusion (I/R) injury in rats.Renal function, renal histopathological changes, plasma P-selectin levels and renal P-selectin protein and mRNA expression were studied in a renal I/R injury rat models. Biochemical measurement, ELISA, Immunohistochemistry and Nested RT-PCR were used.Renal function insufficiency and renal histopathological damage were much less in the anti-P-selectin mAb-treated group than the saline-treated group. Plasma P-selectin levels were lower and renal P-selectin protein and mRNA expression were down-regulated in the former group.Anti-P-selectin mAb might be an efficient approach for the treatment of renal I/R injury.
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