ABSTRACT
Hypertrophic Scar (HS) is a complicated fibrotic disease. In addition, its pathogenesis is still to be further explored. Long non-coding RNAs (lncRNAs) have been proved to be participated in multiple diseases, including HS. However, the role of lncRNA TUG1 in HS remains unclear. The expression level of RNA and protein in cells were detected by q-PCR and western blot, respectively. MTT assay was performed to test the cell proliferation. Cell migration was detected by transwell assay. Cell apoptosis was measured by flow cytometry. Dual luciferase report assay and RNA pull down were used to verify the relationship between TUG1, miR-27b-3p and TAK1.TUG1 and TAK1 were upregulated in HS, while miR-27b-3p was downregulated. Knockdown of TUG1 significantly suppressed the proliferation and migration and induced the apoptosis of HS fibroblasts (HSF). In addition, silencing of TUG1 notably inhibited the extracellular matrix (ECM) biosynthesis in HSF. Overexpression of miR-27b-3p has the same effect on HS as that of TUG1 knockdown. Meanwhile, TUG1 could sponge miR-27b-3p, and TAK1 was the direct target of miR-27b-3p. Furthermore, knockdown of TUG1 significantly suppressed the fibrosis in HS via miR-27b-3p/TAK1/YAP/TAZ axis mediation. LncRNA TUG1 promotes the fibrosis in HS via sponging miR-27b-3p and then activates TAK1/YAP/TAZ pathway, which may serve as a potential target for treatment of HS.
Subject(s)
Cell Cycle Proteins/metabolism , Cicatrix, Hypertrophic/pathology , Fibrosis/pathology , Intracellular Signaling Peptides and Proteins/metabolism , MAP Kinase Kinase Kinases/metabolism , RNA, Long Noncoding/genetics , Transcription Factors/metabolism , Apoptosis , Biomarkers/metabolism , Cell Cycle Proteins/genetics , Cell Movement , Cell Proliferation , Cells, Cultured , Cicatrix, Hypertrophic/genetics , Cicatrix, Hypertrophic/metabolism , Fibrosis/genetics , Fibrosis/metabolism , Gene Expression Regulation , Humans , Intracellular Signaling Peptides and Proteins/genetics , MAP Kinase Kinase Kinases/genetics , MicroRNAs/genetics , Prognosis , Transcription Factors/genetics , Transcriptional Coactivator with PDZ-Binding Motif ProteinsABSTRACT
The aim of the present study was to prepare a novel CD133 aptamer modified DTX liposome system and investigate its characteristics in vitro and in vivo studies. In this study, the CD133-DTX LP was prepared by the thin-film hydration method and with the particle size of 100-120 nm. The TEM photomicrographs were smooth, sub-spherical in shape and aggregated to form small clusters. In vitro, a relatively slower DTX release profile was observed in CD133-DTX LP due to the presence of CD133 aptamers on the outer surface which might hinder the drug release. The drug release mechanism fit well with the Higuchi equation better. In cytotoxicity study, CD133 aptamers modified DTX LP significantly decreased cell proliferation and improved the therapeutic efficiency. In vivo imaging result indicated that CD133-DTX LP had very good tumour targeting ability. In vivo antitumour activity indicated that the CD133-DTX LP showed a significant antitumour activity in A549 tumour mice, with a very low systemic toxicity.
Subject(s)
AC133 Antigen , Docetaxel , Drug Delivery Systems/methods , Lung Neoplasms/drug therapy , A549 Cells , AC133 Antigen/chemistry , AC133 Antigen/pharmacokinetics , AC133 Antigen/pharmacology , Animals , Docetaxel/chemistry , Docetaxel/pharmacokinetics , Docetaxel/pharmacology , Humans , Liposomes , Lung Neoplasms/metabolism , Lung Neoplasms/pathology , Rabbits , Surface PropertiesABSTRACT
Development of chemoresistance remains a major hurdle for triple negative breast cancer treatment. Previous studies suggest that CD44+/CD24- cells, subpopulation of cancer stem cells with self-renewing and tumor-initiating capacities, are partly responsible for chemoresistance and therapeutic failure of triple negative breast cancer. Therefore, novel agents that target cancer stem cells (CSCs) may improve the clinical outcome. KIF11 (kinesin family member 11), overexpressed in many cancer cells, is a molecular motor protein that plays essential role in mitosis. In this study, we assess its role in docetaxel resistant triple negative breast cancer (TNBC). We found that the expression of KIF11 was significantly increased in CD44+/CD24- subpopulation of docetaxel resistant TNBC cells. Knockdown of KIF11 resulted in a significant decrease in the percentage of CSCs and mammosphere formation. KIF11 knockdown also inhibits cell growth and induces cell cycle G2/M arrest followed by cell mitosis and apoptosis. Further docetaxel resistant TNBC xenograft models demonstrated that KIF11 inhibitor exerts growth inhibitory effect in vivo. Of note, we also found that KIF11 was highly expressed in TNBC and its expression was correlated with shorter disease free survival time. All these data indicate that KIF11 is critical for proliferation and self-renewal in TNBC tumor cells in vitro and in vivo, suggesting that KIF11 may be a promising therapeutic target for treating chemoresistant TNBC.
ABSTRACT
This study investigated the impact of four environmental factors on the recruitment of cyanobacteria from bottom sediments in the eutrophic Shanzi Reservoir. Temperature and light were identified as the key determinants for the recruitment of Microcystis and Oscillatoria. Cyanobacteria became dominant at higher temperature (20 °C) and light intensity (2000â lx) and Microcystis and Oscillatoria were the major species. Detailed recruitment simulation undertaken with the respective gradients of temperature and light suggested that both Microcystis and Oscillatoria are temperature sensitive and that their critical temperature point was 10 °C. However, distinct light impacts were observed only on Microcystis. The recruitment of Oscillatoria was light independent, whereas Microcystis had a positive relationship with light intensity. Physical disturbance promoted Microcystis recruitment and also affected the structure of the recruited cyanobacterial community at the water-sediment interface, based on quantitative polymerase chain reaction (qPCR) and phylogenetic analysis.
Subject(s)
Geologic Sediments/microbiology , Microcystis/isolation & purification , Oscillatoria/isolation & purification , Water Microbiology , China , DNA, Bacterial/genetics , Eutrophication , Light , Microcystis/genetics , Oscillatoria/genetics , Phylogeny , Polymerase Chain Reaction , RNA, Ribosomal, 16S/genetics , Temperature , Water SupplyABSTRACT
Primary investigation of Platymonas subcordiformis was performed with laser scanning microscopic technology. Both autofluorescence spectra and autofluorescence imaging of Platymonas subcordiformis were achieved by one photon excitation of 488 nm Ar+ laser. Autofluorescence images revealed cup-shaped object inside the Platymonas subcordiformis cell, and the corresponding fluorescence peak located at 682 nm was attributed to chloroplast. Moreover, in single channel mode, there existed a round-shaped object in chloroplast center, showing strong fluorescence by 800 nm femtosecond laser excitation. Also, the authors obtained both the individual cup-shaped chloroplast and round-shaped object image and the overlaid images by using dual-channel mode. Meanwhile, six main fluorescence peaks were found. Single photon excitation can be used to obtain autofluorescence image and autofluorescence spectra of Platymonas subcordiformis, while two photon excitation combined with multitrack mode and Lambda mode can be used not only to observe internal structure, but also for the analysis of existence of biological and chemical substances with higher sensitivity than single photon excitation. LSCM technique can be a promising tool for rapid, convenient, real-time and effective investigation in ocean algae.
Subject(s)
Chlorophyta , Fluorescence , Chloroplasts , Lasers , Light , Microscopy, Confocal , Microscopy, Fluorescence, Multiphoton , PhotonsABSTRACT
Autofluorescence spectra and optical imaging of Platymonas subcordiformis after irradiation of diode laser were observed via laser scanning confocal microscopy (LSCM). With 488 nm Ar(+) laser excitation, the horizontal and vertical dimensions of a cup-shaped chloroplast of the irradiation group increased about 10% compared with the control group. The fluorescence spectra were similar between irradiation group and control group with a maximum fluorescence band around 682 nm, whereas the former has a higher intensity. Image of a small circular substance with stronger two-photon autofluorescence (TPA) was obtained when using two-photon excitation wavelength of 800 nm in single-channel mode. Further analysis by the 800 nm excitation based on two independent-channels mode showed an emission band of the small circular substance around 376-505 nm, which corresponded to the eyespot of P. subcordiformis. In lambda scanning mode, with two-photon wavelength of 800 nm excitation, six fluorescence peaks that are located at 465, 520, 560, 617, 660 and 680 nm were observed; the fluorescence intensity of the irradiation group was higher than that of the control group, especially at 520, 560 and 617 nm. As a conclusion, diode laser irradiation can promote chloroplast growth of P. subcordiformis cells in the form of expanding area and the increasing content of protein, phospholipids and chlorophyll. LSCM, especially TPA imaging based on femtosecond laser excitation, provides a nondestructive, real-time and accurate method to study changes of living algal cells under laser irradiation and other environmental factors.
Subject(s)
Chlorophyta/radiation effects , Lasers, Semiconductor , Microscopy, Confocal/methods , Microscopy, Fluorescence, Multiphoton/methods , Spectrometry, Fluorescence/methods , Chlorophyta/physiology , Chlorophyta/ultrastructure , Microscopy, Confocal/instrumentation , Microscopy, Fluorescence, Multiphoton/instrumentation , Pigments, Biological , Spectrometry, Fluorescence/instrumentationABSTRACT
FT-Raman spectroscopy was adopted to investigate the Raman spectrum of Platymonas subcordiformis. The result shows that the optimal experiment condition is making sample lose solvent with centrifuge, exciting laser power set at 360 mW and accumulating 70 times. The main peaks of the spectra are 381-432, 552-556, 611-613, 710, 873, 953-964, 1 108-1119, 1457, 1523-1527 and 2986 cm(-1). They belong to protein, insaturation acid and ester, etc, which are the main composition of Platymonas subcordiformis. The precise measurements of algae Raman spectra could be used for developing a new optical taxonomic methodology to distinguish between different algae species, and a rapid, non-destructive detection way of stress effects.
