[Show abstract][Hide abstract] ABSTRACT: Recent studies have demonstrated the critical roles of micro-RNAs in tumorigenesis and tumor progression. Here, we describe the regulation and function of miR-490-5p in bladder cancer.
Paired tissue samples were collected from bladder cancer patients (n = 20). Real-time PCR revealed that miR-490-5p expression was significantly down-regulated in human bladder cancer tissues and cells. Also there was an inverse relationship between the expression level of miR-490-5p and the pathological grade of bladder cancer. Western blotting was performed to detect the expression levels of c-FOS and TET1 in 6 matched tumor tissue samples and 4 bladder cell lines. Furthermore, to better understand the underlying mechanisms of miR-490-5p, we conducted gain and loss of function analysis by transfecting bladder cancer T24 cells with chemically synthesized miR-490-5p mimics and inhibitor, respectively.
We found that overexpression of miR-490-5p in T24 cells could inhibit cell proliferation and invasion and induce cell apoptosis. Conversely, suppression of miR-490-5p expression induced cell proliferation and invasion, while it inhibited cell apoptosis. In addition, our bioinformatics prediction and experimental data showed that c-FOS was a potential target of miR-490-5p. The expression level of c-FOS was significantly decreased after miR-490-5p overexpression and significantly increased after miR-490-5p suppression, indicating that c-FOS was a target of miR-490-5p.
These findings suggest that miR-490-5p is a novel tumor suppressor, contributing to the carcinogenesis of bladder cancer by targeting c-FOS.
Preview · Article · Jun 2015 · Archives of Medical Science
[Show abstract][Hide abstract] ABSTRACT: The present study aimed to investigate the effect of hydrogen sulfide (H2S) on kidney injury induced by urinary-derived sepsis. Rabbits were randomly divided into control, sham, sepsis, NaHS 2.8 μmol/kg and NaHS 8.4 μmol/kg groups, with six rabbits in each group. Upper urinary tract obstruction and acute infection was induced to establish the sepsis model. Blood was collected to carry out a white blood cell (WBC) count, and creatinine (Cr) and blood urea nitrogen (BUN) analysis. Morphological changes were observed by hematoxylin and eosin (H&E) staining and transmission electron microscopy. Immunohistochemical staining was used to detect the expression levels of tumor necrosis factor (TNF)-α, interleukin (IL)-10 and nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB). Cystathionine-γ-lyase (CSE) activity was measured by the spectrophotometric methylene blue method and the blood H2S concentration was measured by deproteinization. WBC, Cr and BUN levels were significantly elevated in the sepsis group compared with those in the control group (P<0.05). Following treatment with NaHS, the WBC, Cr and BUN levels were significantly decreased in the NaHS groups compared with those in the sepsis group (P<0.05). The pathological features of kidney injury were also alleviated by NaHS. In the sepsis group, the levels of TNF-α, IL-10 and NF-κB were significantly increased compared with those in the control group (P<0.05). In the NaHS groups, the TNF-α and NF-κB levels were significantly reduced whereas the IL-10 level was significantly increased compared with the respective levels in the sepsis group (P<0.05). The H2S concentration was significantly decreased in the sepsis group and this reduction was attenuated in the NaHS groups (P<0.05). Furthermore, the NaHS 8.4 μmol/kg dose revealed a more potent effect than the NaHS 2.8 μmol/kg dose. Thus, exogenous H2S reduced kidney injury from urinary-derived sepsis by decreasing the levels of NF-κB and TNF-α, and increasing the level of IL-10.
Preview · Article · Aug 2014 · Experimental and therapeutic medicine
[Show abstract][Hide abstract] ABSTRACT: The aim of the present study was to determine the efficacy of ulinastatin (UTI) for the treatment of sepsis and to investigate the associated molecular mechanisms. Twenty‑four male rabbits were randomly divided into 4 groups, the normal, sham, sepsis model and UTI groups, each containing 6 rabbits. Serum levels of interleukin (IL)‑10 and tumor necrosis factor‑α (TNF‑α) were measured by enzyme‑linked immunosorbent assay (ELISA). Liver, kidney and lung tissues were stained with hematoxylin and eosin (H&E) 36 h after sacrifice and morphological changes were observed under an optical microscope. The expression levels of IL‑10 and TNF‑α proteins in rabbit kidney tissue in each group were determined by immunohistochemical detection and western blot analysis. ELISA results indicated that, compared with the sepsis model, IL‑10 levels were significantly higher in the UTI treatment group (183.91±11.521 pg/ml) at 36 h (P=0.000), while serum TNF‑α concentration decreased significantly in the UTI treatment group (31.637±2.770 pg/ml; P=0.000). Results of western blot analysis were consistent with the immunohistochemistry, indicating that UTI upregulates IL‑10 and downregulates TNF‑α levels. In the current study, UTI was demonstrated to effectively treat urinary sepsis and alleviate the inflammatory response in tissues. These effects were mediated by the upregulation of IL‑10 and downregulation of TNF‑α levels.
Preview · Article · May 2013 · Molecular Medicine Reports
[Show abstract][Hide abstract] ABSTRACT: β-elemene, a non-cytotoxic antitumor reagent, inhibits the growth, proliferation and DNA synthesis of multiple types of malignant cells, resulting in the apoptosis or suppressed vascularization of tumors. β-elemene is also able to enhance the immunogenicity of the tumor cells and ameliorate systematic cellular immunity in the tumor‑bearing body. Moreover, β-elemene has the advantages of high efficiency, safety and low possibility of drug tolerance over other antitumor agents used in antitumor treatment. Therefore, β-elemene has great potential in clinical applications. However, the mechanism of β-elemene antitumor activity is largely unknown. The aim of this study was to investigate whether β-elemene is able to repress the proliferation of T24 bladder carcinoma cells through regulation of the expression of the tumor suppressor gene, Smad4. Results of a methylthiazolyl tetrazolium (MTT) assay indicated that the proliferation of T24 cells was repressed by β-elemene in a time- and concentration‑dependent manner. The lowest concentration of β-elemene to inhibit cell survival by >50% was determined using IC50 software. Microscopic observation also demonstrated the potential of β-elemene to induce the apoptosis of cancer cells. Western blot and RT-PCR analyses revealed that the expression of the Smad4 protein and mRNA was upregulated by treatment with β-elemene. Our results revealed that β-elemene was able to upregulate the expression of Smad4 in tumor cells to inhibit the proliferation of these cells.
No preview · Article · Nov 2012 · Molecular Medicine Reports
[Show abstract][Hide abstract] ABSTRACT: β-elemene, a non-cellular antineoplastic agent, may be used to effectively inhibit the growth and proliferation of various types of tumor cells by inhibiting the nucleic acid synthesis or inducing their apoptosis and differentiation. The aim of this study was to investigate the expression, as well as the effects, of Mta-1, survivin and Bcl-xL in T24 bladder cancer cells following β-elemene treatment. The expression of the three proteins in T24 cells following β-elemene treatment was analyzed by immunocytochemistry staining and western blot analysis. The survival rate and apoptosis of T24 cells following β-elemene treatment was detected by MTT assay and TUNEL staining. We analyzed the internal corelations between apoptosis-associated genes, tumor metastasis-associated genes (cancer genes) and cell apoptosis, and investigated the mechanism of action by which β-elemene induces the apoptosis of T24 cells at a molecular level. These results provide scientific evidence for further study on the anticancer effect of β-elemene in carcinoma of the urinary bladder. In this study, it is shown that β-elemene downregulates the expression of survivin, Bcl-xL and Mta-1 in tumor cells. The apoptosis of T24 cells is dependent on the dosage and length of incubation time of β-elemene.
Preview · Article · Aug 2012 · Molecular Medicine Reports
[Show abstract][Hide abstract] ABSTRACT: FeIII porphyrin complexes as bioinspired catalysts exhibit remarkable catalytic activity for sulfides oxidation near the natural conditions, i.e. at room temperature, in the protonic solvent and with psychological oxidant H2O2 aqueous in the absence of other chemical assistance. Various sulfur substrates existing in petroleum were examined including refractory dibenzothiophene. Reaction conditions affecting the conversion of sulfides and the stability of catalyst were investigated. The results showed that polar and protonic solvent can facilitate this oxidation and the catalyst's life-span was closely related with temperature variations. FeIII porphyrin, substituted on the phenyl groups with a chlorine atom, displayed higher efficiency than no substituted. The conversions calculated from the GC data are 100, 99.81, 99.78 and 77.05 wt % for di-n-butyl sulfide, thioanisole, diphenyl sulfide and dibenzothiophene respectively after a typical procedure.
[Show abstract][Hide abstract] ABSTRACT: The purpose of this study was to explore the functional role and mechanism of miR-155 in the development of renal cell carcinoma (RCC). miR-155 expression was quantified in renal cancers, matched adjacent non‑tumor tissues and renal cell lines using quantitative real-time PCR (RT-PCR). Cell proliferation, apoptosis and migratory activity were measured following suppression of miR-155 expression by antisense oligonucleotides. miR-155 targets were scanned using target prediction programs. Following the inhibition of miR-155, target gene expression was detected by western blotting. The expression of miR-155 was upregulated in clear cell RCC (ccRCC) tissue and renal cancer cell lines. The suppression of miR-155 inhibited cell proliferation and migratory activity and induced apoptosis in renal cancer cells. The suppressor gene suppressor of cytokine signaling (SOCS-1) and BACH1 were predicted as potential target genes by bioinformatics analysis. The suppression of miR-155 inhibited BACH1 protein expression. miR-155 may function as an oncogene by targeting BACH1. Thus, the inhibition of miR-155 may be an effective way to treat RCC.
Preview · Article · Apr 2012 · Molecular Medicine Reports
[Show abstract][Hide abstract] ABSTRACT: The development of effective therapies for the prevention of colorectal cancer (CRC) liver metastasis is of great importance. Recently, chitosan (CS) nanoparticles have been utilized as carriers of interluekin-12 (IL-12) administered locally to deliver therapeutic proteins and genes. In this study, we encapsulated IL-12 by incorporation using tripolyphosphate (TPP) as the coacervated crosslinking agent to form CS-TPP/IL-12 nanoparticles. We further characterized the association efficiency, rate of release, liver-targeting, and toxicity, which were predominantly dependent on the factors of particle size, zeta potential, pH of solution, and whether or not modified with TPP. Systemic delivery of CS-TPP/IL-12 nanoparticles significantly reduced the number and volume of CRC liver metastasis foci compared to the CS-TPP treated mouse group. Although delivery of IL-12 alone also inhibited the number of CRC liver metastasis observed, further study of the change in hepatic metastasis volume demonstrated no significant differences between the groups treated with CS-TPP or IL-12 alone. Mechanistically, CS-TPP nanoparticles blocked the toxicity of IL-12 and induced infiltration of NK cells and some T cells, which are most likely the effector cells that mediate tumor metastasis inhibition during CS-TPP/IL-12 immunotherapy. The results obtained from this study demonstrate the potential benefit of using chitosan modification technology as a cytokine delivery system for the successful prevention of CRC liver metastasis by exploiting liver immunity.
[Show abstract][Hide abstract] ABSTRACT: Numerous microRNAs (miRNAs) play crucial roles in cancer development. In this study, we report that hsa-miR-96 is expressed at higher levels in human bladder urothelial carcinomas compared to normal tissues. We found that hsa-miR-96 increased invasion and differentiation of human bladder T24 cells and promoted their growth. Down‑regulation of hsa-miR-96 significantly affected the phenotype of bladder cancer T24 cells. The mRNA and protein levels of insulin receptor substrate 1 (IRS1) and MAP4K1 were significantly reduced in cells transfected with the hsa-miR-96 inhibitor when compared with levels in cells transfected with the empty plasmid vector or the negative control miRNA inhibitor. Altogether, these results suggest that hsa-miR-96 may affect the growth of bladder cancer cells by up-regulating IRS1 and MAP4K1 levels, functioning as a promising diagnostic marker in human bladder urothelial carcinomas.
Preview · Article · Jan 2012 · Molecular Medicine Reports
[Show abstract][Hide abstract] ABSTRACT: It was the aim of this study to explore the effects of 3-(5'-hydroxymethyl-2'-furyl)-l-benzyl indazole (YC-1) on transcription activity, cell proliferation and apoptosis of hypoxic human bladder transitional carcinoma cells (BTCC), mediated by hypoxia-inducible factor 1α (HIF-1α).
BTCC cell line T24 cells were incubated under normoxic or hypoxic conditions, adding different doses of YC-1. The protein expression of HIF-1α and HIF-1α-mediated genes was detected by Western blotting. RT-PCR was used to detect HIF-1α mRNA expression. Cell proliferation, apoptosis and migration activity were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry and transwell migration assay. The cells were pretreated by two ERK/p38 MAPK pathway-specific inhibitors, PD98059 or SB203580, and then incubated with YC-1 treatment under hypoxic condition. HIF-1α protein expression was detected by Western blotting.
Hypoxic T24 cells expressed a higher level of HIF-1α, vascular endothelial growth factor, matrix metalloproteinases-2, B-cell lymphoma/leukemia-2 protein and HIF-1α mRNA compared with normoxic controls, in which the above-mentioned expression was downregulated by YC-1 in a dose-dependent manner. Cell proliferation and migration activity were inhibited while apoptosis was induced by YC-1 under hypoxic condition. Moreover, YC-1-downregulated HIF-1α expression was reversed by PD98059 and SB203580, respectively.
YC-1 inhibits HIF-1α and HIF-1α-mediated gene expression, cell proliferation and migration activity and induces apoptosis in hypoxic BTCC. The ERK/p38 MAPK pathway may be involved in YC-1-mediated inhibition of HIF-1α.
No preview · Article · Jan 2012 · Urologia Internationalis