ABSTRACT
BACKGROUND AND OBJECTIVES@#Recent studies have examined the structure-function relationship of high-density lipoprotein (HDL). This study aimed to identify and rank HDL-associated proteins involved in several biological function of HDL.@*METHODS@#HDLs isolated from 48 participants were analyzed. Cholesterol efflux capacity, effect of HDL on nitric oxide production, and vascular cell adhesion molecule-1 expression were assessed. The relative abundance of identified proteins in the highest vs. lowest quartile was expressed using the normalized spectral abundance factor ratio.@*RESULTS@#After adjustment by multiple testing, six proteins, thyroxine-binding globulin, alpha-1B-glycoprotein, plasma serine protease inhibitor, vitronectin, angiotensinogen, and serum amyloid A-4, were more abundant (relative abundance ratio ≥2) in HDLs with the highest cholesterol efflux capacity. In contrast, three proteins, complement C4-A, alpha-2-macroglobulin, and immunoglobulin mu chain C region, were less abundant (relative abundance ratio <0.5). In terms of nitric oxide production and vascular cell adhesion molecule-1 expression, no proteins showed abundance ratios ≥2 or <0.5 after adjustment. Proteins correlated with the functional parameters of HDL belonged to diverse biological categories.@*CONCLUSIONS@#In summary, this study ranked proteins showing higher or lower abundance in HDLs with high functional capacities and newly identified multiple proteins linked to cholesterol efflux capacity.
ABSTRACT
BACKGROUND AND OBJECTIVES: Recent studies have examined the structure-function relationship of high-density lipoprotein (HDL). This study aimed to identify and rank HDL-associated proteins involved in several biological function of HDL.METHODS: HDLs isolated from 48 participants were analyzed. Cholesterol efflux capacity, effect of HDL on nitric oxide production, and vascular cell adhesion molecule-1 expression were assessed. The relative abundance of identified proteins in the highest vs. lowest quartile was expressed using the normalized spectral abundance factor ratio.RESULTS: After adjustment by multiple testing, six proteins, thyroxine-binding globulin, alpha-1B-glycoprotein, plasma serine protease inhibitor, vitronectin, angiotensinogen, and serum amyloid A-4, were more abundant (relative abundance ratio ≥2) in HDLs with the highest cholesterol efflux capacity. In contrast, three proteins, complement C4-A, alpha-2-macroglobulin, and immunoglobulin mu chain C region, were less abundant (relative abundance ratio <0.5). In terms of nitric oxide production and vascular cell adhesion molecule-1 expression, no proteins showed abundance ratios ≥2 or <0.5 after adjustment. Proteins correlated with the functional parameters of HDL belonged to diverse biological categories.CONCLUSIONS: In summary, this study ranked proteins showing higher or lower abundance in HDLs with high functional capacities and newly identified multiple proteins linked to cholesterol efflux capacity.
Subject(s)
Amyloid , Angiotensinogen , Atherosclerosis , Cardiovascular Diseases , Cholesterol , Complement System Proteins , Immunoglobulin mu-Chains , Lipoproteins , Nitric Oxide , Plasma , Proteomics , Serine Proteases , Thyroxine-Binding Globulin , Vascular Cell Adhesion Molecule-1 , VitronectinABSTRACT
PURPOSE: Our aim of research is to find novel genes that overexpressed in various samples such as cell lines and tissues that infinitely proliferate; so-called immortalized cells, cancer cells and tissues. In this study, we obtained a gene from immortalized cell line (WI-38 VA13) then identified it from various cell lines and human lung tissues. MATERIALS AND METHODS: Using suppressive subtractive hybridization (SSH) method, we obtained many genes and selected a novel gene of them. And then the novel gene fragment was amplified by PCR and ligated in T easy vector for sequencing. And finally we found a differentially expressed gene in cell lines and tissues when it was performed by reverse transcriptase-PCR (RT-PCR). RESULTS: As the result of transformation of genes that we gained using SSH, we obtained about 150 clones. And then we certificated several genes by DNA prep and confirmed it by sequencing. We examined whether the gene sequences are novel or known genes by genome homology search and we confirmed the gene expressions by RT-PCR. As a result, we identified a differentially overexpressed gene (named "clone 58") in immortalized cells, cancer cell lines and lung squamous cell carcinomas. CONCLUSION: The "clone 58" mRNA was significantly up-regulated in various human cell lines and also human lung cancer tissues compared to the normal. We suppose that this gene can carry out a specific role related to the induction of cancer and/or the mechanism of the changeover of a normal cell to an immortalized cell. In short, the discovery of our gene has an importance in the point that they are thought to have a connection with immortalization and carcinogenesis of human cells and tissues.