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
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
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.
