10 research outputs found
Morphological change of SMMC-7721 cells and HUVECs in a 3D co-culture model.
<p>(A) SMMC-7721 cells and HUVECs were seeded and cultured in Matrigel. Morphogenesis was observed by using an inverted phase-contrast microscope. (B) These 3D-cultured cell sheets were fixed and cut into sections which were stained with anti CD147 (green) and KDR/Flk (red) antibodies, and DAPI (blue), respectively, and observed using a laser scanning confocal microscope.</p
Knock-down of CD147 in SMMC-7721 cells decreased the levels of VEGF and IGF-I in TCMs under normoxia and hypoxia conditions.
<p>All figures are a representation of three trials. The concentrations of VEGF (A), IGF-I (B), bFGF (C) and EGF (D) in TCMs were determined by ELISA analysis. *<i>P</i><0.05, <sup>#</sup><i>P</i>>0.05, compared to the control level.</p
The angiogenic phenotype was inhibited by knock-down of CD147 in either SMMC-7721 cells (TC) or endothelial cells (EC) in a 3D co-culture model.
<p>(A) CD147 expression was analyzed by western blot in SMMC-7721 cells and HUVECs after transfected with CD147 siRNA or control siRNA for 48 h. (B) <i>In vitro</i> tube formation analysis in Matrigel. After SMMC-7721 cells and HUVECs were transfected with CD147 siRNA or control siRNA respectively, SMMC-7721 cells were co-cultured with HUVEC cells for 48 h in Matrigel. Images were acquired at 6, 24 and 48 h, using an inverted microscope (Olympus CKX41) fitted with a 10× phase-contrast objective lens. (C) Semi-quantitative assessment of tube formation was performed by determining the number of branches per field. Results are based on four randomly selected fields and are expressed as the mean ± SD of three independent experiments. Statistical significance was determined by the Student’s t-test. *<i>P</i><0.05, **<i>P</i><0.001, compared with the control level.</p
IGF-I induced CD147 expression in HUVECs and SMMC-7721 cells.
<p>CD147 expression level was determined by qRT-PCR and western blot in HUVECs (A and B) or SMMC-7721 (C and D) which were treated with VEGF, bFGF, EGF and IGF-I (100ng/mL) for 24 h. *<i>P</i><0.05, **<i>P</i><0.0001, compared with the control level.</p
IGF-I induced CD147 expression in dose-dependent fashion in multiple tumor cells.
<p>(A, B) qRT-PCR (A) and Western blot (B) analysis of CD147 expression level in SMMC-7721, HepG-2, A549, and MCF-7 cells in response to stimulation by IGF-I for 24 h.</p
Knock-down of CD147 down-regulated the expression of VEGF, IGF-I and HIF-1α in normoxia and hypoxia conditions.
<p>All figures are a representation of three trials. (A) RT-PCR was performed to examine the transcription levels of CD147, VEGF, IGF-I, HIF-1α and GAPDH in SMMC-7721 cells transfected with CD147 siRNA or control siRNA. (B) Western blots were performed to examine the expression level of CD147, HIF-1α, and β-actin proteins in SMMC-7721 cells transfected with CD147 siRNA or control siRNA. The expression of β-actin was used as internal control. (C) qRT-PCR was performed to examine the transcription levels of these molecular in SMMC-7721 cells transfected with CD147 siRNA or control siRNA. *<i>P</i><0.05, **<i>P</i><0.001, compared with the control level.</p
IGF-I was a specific up-regulator of CD147 expression to promote angiogenesis.
<p>We removed IGF-I from TCMs by using immunoprecipitation and observed the change of proliferation (A) and migration (B) and formation tube-like structures (C and D) of HUVECs response to TCM, IGF-I and siRNA CD147.</p
Solid-State NMR Shows That Dynamically Different Domains of Membrane Proteins Have Different Hydration Dependence
Hydration has a profound influence
on the structure, dynamics,
and functions of membrane and membrane-embedded proteins. So far the
hydration response of molecular dynamics of membrane proteins in lipid
bilayers is poorly understood. Here, we reveal different hydration
dependence of the dynamics in dynamically different domains of membrane
proteins by multidimensional magic angle spinning (MAS) solid-state
NMR (ssNMR) spectroscopy using 121-residue integral diacylglycerol
kinase (DAGK) in 1,2-dimyristoyl-<i>sn</i>-glycero-3-phosphocholine
(DMPC)/1,2-dimyristoyl-<i>sn</i>-glycero-3-phospho-(1′-<i>rac</i>-glycerol) (DMPG) lipid bilayers as a model system. The
highly mobile and immobile domains of DAGK and their water accessibilities
are identified site-specifically by scalar- and dipolar-coupling based
MAS ssNMR experiments, respectively. Our experiments reveal different
hydration dependence of the dynamics in highly mobile and immobile
domains of membrane proteins. We demonstrate that the fast, large-amplitude
motions in highly mobile domains are not triggered until 20% hydration,
enhanced at 20–50% hydration and unchanged at above 50% hydration.
In contrast, motions on submicrosecond time scale of immobile residues
are observed to be independent of the hydration levels in gel phase
of lipids, and at the temperature near gel–liquid crystalline
phase transition, amplitude of whole-molecule rotations around the
bilayer normal is dominated by the fluidity of lipid bilayers, which
is strongly hydration dependent. The hydration dependence of the dynamics
of DAGK revealed by this study provides new insights into the correlations
of hydration to dynamics and function of membrane proteins in lipid
bilayers
NaGdF<sub>4</sub>:Yb<sup>3+</sup>/Er<sup>3+</sup>@NaGdF<sub>4</sub>:Nd<sup>3+</sup>@Sodium-Gluconate: Multifunctional and Biocompatible Ultrasmall Core–Shell Nanohybrids for UCL/MR/CT Multimodal Imaging
Multimodal bioimaging nanoparticles
by integrating diverse imaging ingredients into one system, represent
a class of emerging advanced materials that provide more comprehensive
and accurate clinical diagnostics than conventional contrast agents.
Here monodisperse and biocompatible core–shell nanoparticles,
NaGdF<sub>4</sub>: Yb<sup>3+</sup>/Er<sup>3+</sup>@NaGdF<sub>4</sub>:Nd@sodium-gluconate (termed as GNa-Er@Nd), with about 26 nm in diameter
were successfully prepared by a facile two step reactions in high
boiling solvents, and followed a ligand exchange process with sodium
gluconate. The resulting GNa-Er@Nd nanoparticles were well characterized
by transmission electron microscopy (TEM), X-ray diffraction (XRD),
Fourier transform infrared spectra (FTIR), and zeta potentials. These
nanohybrids present brightly dual-wavelength excited upconversion
luminescence (UCL) under both 980 and 793 nm laser because of the
synergistic effect of Yb<sup>3+</sup>/Er<sup>3+</sup> and Nd<sup>3+</sup>. They also exhibited excellent relaxivity parameters (<i>r</i><sub>1</sub>) in magnetic resonance imaging (MRI) and Hounsfield
units (HU) in X-ray computed tomography (CT) that are comparable to
the clinical contrast agents. Therefore, these small and monodisperse
nanoparticles provide options to construct a unique platform for potential
multimodal UCL/CT/MRI imaging simultaneously
Accessible Method for the Development of Novel Sterol Analogues with Dipeptide-like Side Chains That Act as Neuroinflammation Inhibitors
A number of novel sterol derivatives
with dipeptide-like side chains
were synthesized using an Ugi four-component condensation method and
assayed to test their anti-inflammatory effects in activated microglial
cells. Compound <b>18b</b> ((3<i>S</i>,10<i>R</i>,13<i>S</i>)-<i>N</i>-((<i>R</i>)-1-(<i>tert</i>-butylamino)-1-oxo-3-phenylpropan-2-yl)-3-hydroxy-<i>N</i>,10,13-trimethyl-2,3,4,7,8,9,10,11,12,13,14,15-dodecahydro-1<i>H</i>-cyclopentaÂ[<i>a</i>]Âphenanthrene-17-carboxamide)
was identified as the most potent anti-inflammatory agent in the series
of compounds analyzed. Compound <b>18b</b> markedly inhibited
the expression of proinflammatory factors, including inducible nitric
oxide synthase, interleukin (IL)-6, IL-1β, tumor necrosis factor-α,
and cyclooxygenase-2 in lipopolysaccharide-stimulated microglial cells.
Further studies showed that compound <b>18b</b> significantly
suppressed the transcriptional activity of AP-1 and NF-κB in
activated microglial cells, which was likely mediated by the inhibition
of the p38 MAPK and JNK signal transduction pathways. In addition,
compound <b>18b</b> displayed neuroprotective effects in a microglial-conditioned
medium/neuron coculture and an experimental focal ischemic mouse model