Maintenance of the stemness in CD44+ HCT-15 and HCT-116 human colon cancer cells requires miR-203 suppression

AbstractThe purpose of this study was to isolate cancer stem cells (CSCs, also called tumor-initiating cells, TICs) from established human colorectal carcinoma (CRC) cell lines, characterize them extensively and dissect the mechanism for their stemness. Freshly isolated CD44+ and CD44− cells from the HCT-15 human colon cancer cell line were subjected to various analyses. Interestingly, CD44+ cells exhibited higher soft agar colony-forming ability and in vivo tumorigenicity than CD44− cells. In addition, the significant upregulation of the protein Snail and the downregulation of miR-203, a stemness inhibitor, in CD44+ cells suggested that this population possessed higher invasion/metastasis and differentiation potential than CD44− cells. By manipulating the expression of CD44 in HCT-15 and HCT-116 cells, we found that the levels of several EMT activators and miR-203 were positively and negatively correlated with those of CD44, respectively. Further analyses revealed that miR-203 levels were repressed by Snail, which was shown to bind to specific E-box(es) present in the miR-203 promoter. In agreement, silencing miR-203 expression in wild-type HCT-116 human colon cancer cells also resulted in an increase of their stemness. Finally, we discovered that c-Src kinase activity was required for the downregulation of miR-203 in HCT-15 cells, which was stimulated by the interaction between hyaluronan (HA) and CD44.Taken together, CD44 is a critical molecule for modulating stemness in CSCs. More importantly, we show for the first time that the downregulation of miR-203 by HA/CD44 signaling is the main reason for stemness-maintenance in colon cancer cells

Bioequivalence Evaluation of Two Formulations of Celecoxib 200 mg Capsules in Healthy volunteers by using a validated LC/MS/MS method

The bioequivalence study to compare a new formulation of celecoxib to its reference formulation was designed as an open-label, randomized, single-dose, two-way crossover, comparative bioavailability study by using a validated LC/MS/MS method. In order to determine the plasma concentrations of celecoxib, a sensitive LC/MS/MS method was developed. The method was validated to possess adequate specificity, linearity, precision, accuracy and stability. The linearity of calibration curve was assessed between the concentration intervals (5–2000 ng/mL) with a correlation coefficient over 0.999. Regarding pharmacokinetic investigation, the mean celecoxib AUC0-t values from the test and reference drug formulations were 7360.44 ± 1714.14 h•ng/mL and 7267.48 ± 2077.68 h•ng/mL, respectively, and the corresponding AUC0-∞ values were 8197.45 ± 2040.31 h•ng/mL and 7905.54 ± 2286.12 h•ng/mL, respectively. The Cmax of the test and reference drugs was 705.30 ± 290.63 ng/mL and 703.86 ± 329.91 ng/mL, respectively, and the corresponding Tmax was 3.4 ± 1.6 h and 2.9 ± 1.4 h. Lastly, the T1/2 values of the test and reference drugs were 13.9 ± 7.9 h and 12.9 ± 7.7 h, respectively. The 90% confidence intervals for AUC0-t, AUC0-∞, and Cmax were 97.00-108.85, 98.01-112.09, and 93.20-116.13, respectively, satisfying the bioequivalence criteria of 80-125% range. In conclusion, these results demonstrated that the bioequivalence of two formulations of celecoxib was established successfully by utilizing present developed LC/MS/MS method

Asian Society of Gynecologic Oncology International Workshop 2014

published_or_final_versio

Emerging Two-Dimensional Crystallization of Cucurbit[8]uril Complexes: From Supramolecular Polymers to Nanofibers.

The binding of imidazolium salts to cucurbit[8]uril, CB[8], triggers a stepwise self-assembly process with semiflexible polymer chains and crystalline nanostructures as early- and late-stage species, respectively. In such a process, which involves the crystallization of the host-guest complexes, the guest plays a critical role in directing self-assembly toward desirable morphologies. These include platelet-like aggregates and two-dimensional (2D) fibers, which, moreover, exhibit viscoelastic and lyotropic properties. Our observations provide a deeper understanding of the self-assembly of CB[8] complexes, with fundamental implications in the design of functional 2D systems and crystalline materials.EPSRC (reference no. EP/ G060649/1), ERC Starting Investigator Grant (project no. 240629, ASPiRe) Next Generation Fellowship from the Walters-Kundert Foundation. MINE- CO, the FSE and the FEDER for funding through projects RYC-2015-18471 (Ramoń y Cajal program) and CTQ2017- 84087-R. Royal Society University Research Fellowship UF160152. EPSRC CDT in Nanoscience and Nanotechnology (NanoDTC), grant number EP/L015978/1

Identification of novel candidate target genes, including EPHB3, MASP1 and SST at 3q26.2–q29 in squamous cell carcinoma of the lung

<p>Abstract</p> <p>Background</p> <p>The underlying genetic alterations for squamous cell carcinoma (SCC) and adenocarcinoma (AC) carcinogenesis are largely unknown.</p> <p>Methods</p> <p>High-resolution array- CGH was performed to identify the differences in the patterns of genomic imbalances between SCC and AC of non-small cell lung cancer (NSCLC).</p> <p>Results</p> <p>On a genome-wide profile, SCCs showed higher frequency of gains than ACs (<it>p </it>= 0.067). More specifically, statistically significant differences were observed across the histologic subtypes for gains at 2q14.2, 3q26.2–q29, 12p13.2–p13.33, and 19p13.3, as well as losses at 3p26.2–p26.3, 16p13.11, and 17p11.2 in SCC, and gains at 7q22.1 and losses at 15q22.2–q25.2 occurred in AC (<it>P </it>< 0.05). The most striking difference between SCC and AC was gains at the 3q26.2–q29, occurring in 86% (19/22) of SCCs, but in only 21% (3/14) of ACs. Many significant genes at the 3q26.2–q29 regions previously linked to a specific histology, such as EVI1,<it>MDS1, PIK3CA </it>and <it>TP73L</it>, were observed in SCC (<it>P </it>< 0.05). In addition, we identified the following possible target genes (> 30% of patients) at 3q26.2–q29: <it>LOC389174 </it>(3q26.2),<it>KCNMB3 </it>(3q26.32),<it>EPHB3 </it>(3q27.1), <it>MASP1 </it>and <it>SST </it>(3q27.3), <it>LPP </it>and <it>FGF12 </it>(3q28), and <it>OPA1</it>,<it>KIAA022</it>,<it>LOC220729</it>, <it>LOC440996</it>,<it>LOC440997</it>, and <it>LOC440998 </it>(3q29), all of which were significantly targeted in SCC (<it>P </it>< 0.05). Among these same genes, high-level amplifications were detected for the gene, <it>EPHB3</it>, at 3q27.1, and <it>MASP1 </it>and <it>SST</it>, at 3q27.3 (18, 18, and 14%, respectively). Quantitative real time PCR demonstrated array CGH detected potential candidate genes that were over expressed in SCCs.</p> <p>Conclusion</p> <p>Using whole-genome array CGH, we have successfully identified significant differences and unique information of chromosomal signatures prevalent between the SCC and AC subtypes of NSCLC. The newly identified candidate target genes may prove to be highly attractive candidate molecular markers for the classification of NSCLC histologic subtypes, and could potentially contribute to the pathogenesis of the squamous cell carcinoma of the lung.</p

IL-12p40 Homodimer Ameliorates Experimental Autoimmune Arthritis

IL-23 is the key cytokine that induces the expansion of Th17 cells. It is composed of p19 and p40 subunits of IL-12. The p40 subunit binds competitively to the receptor of IL-23 and blocks its activity. Our aim was to assess the preventive and therapeutic effect of the IL-12p40 homodimer (p40)(2) subunit in autoimmune arthritis animal models. In the current study, using IL-1R antagonist-knockout mice and a collagen-induced arthritis model, we investigated the suppressive effect of (p40)(2) on inflammatory arthritis. We demonstrated that the recombinant adenovirus-expressing mouse (p40)(2) model prevented the development of arthritis when given before the onset of arthritis. It also decreased the arthritis index and joint erosions in the mouse model if transferred after arthritis was established. (p40)(2) inhibited the production of inflammatory cytokines and Ag-specific T cell proliferation. It also induced CD4(+)CD25(+)Foxp3 regulatory T (Treg) cells in vitro and in vivo, whereas the generation of retinoic acid receptor-related organ receptor gamma t and Th17 cells was suppressed. The induction of Treg cells and the suppression of Th17 cells were mediated via activated STAT5 and suppressed STAT3. Our data suggest that (p40)(2) suppressed inflammatory arthritis successfully. This could be a useful therapeutic approach in autoimmune arthritis to regulate the Th17/Treg balance and IL-23 signaling.1156Ysciescopu

High-fidelity spin and optical control of single silicon-vacancy centres in silicon carbide

Scalable quantum networking requires quantum systems with quantum processing capabilities. Solid state spin systems with reliable spin–optical interfaces are a leading hardware in this regard. However, available systems suffer from large electron–phonon interaction or fast spin dephasing. Here, we demonstrate that the negatively charged silicon-vacancy centre in silicon carbide is immune to both drawbacks. Thanks to its 4A2 symmetry in ground and excited states, optical resonances are stable with near-Fourier-transform-limited linewidths, allowing exploitation of the spin selectivity of the optical transitions. In combination with millisecond-long spin coherence times originating from the high-purity crystal, we demonstrate high-fidelity optical initialization and coherent spin control, which we exploit to show coherent coupling to single nuclear spins with ∼1 kHz resolution. The summary of our findings makes this defect a prime candidate for realising memory-assisted quantum network applications using semiconductor-based spin-to-photon interfaces and coherently coupled nuclear spins