RESUMO
The rho-associated coiled-coil protein kinase 2 (ROCK2) gene has been suggested to associate with general hypertension and is therefore a plausible functional candidate gene for pre-eclampsia. ROCK2 maps to chromosome 2p25, which we have implicated previously in a linkage study of pre-eclampsia. We have re-sequenced exons and putative promoter region of ROCK2 in up to 30 pre-eclampsia patients and 22 controls and genotyped putative functional single-nucleotide polymorphisms (SNPs) as well as tagging SNPs from HapMap in a Finnish case-control data set-340 affected and 357 matched control individuals-for a genetic association study of ROCK2 in pre-eclampsia. Even though several new SNPs were discovered, we did not detect significant allelic or haplotypic association between ROCK2 and pre-eclampsia. We assessed ROCK2 expression in placentas by microarray analysis, but no significant expression differences were observed when comparing preeclamptic and normotensive pregnancies. We conclude that common genetic variation in ROCK2 is unlikely to make a major contribution to the risk of pre-eclampsia, but cannot exclude the possibility of having missed non-coding functional variants or rare coding variants.
Assuntos
Alelos , Predisposição Genética para Doença/genética , Pré-Eclâmpsia/genética , Quinases Associadas a rho/genética , Feminino , Finlândia , Genótipo , Humanos , Análise de Sequência com Séries de Oligonucleotídeos , Polimorfismo de Nucleotídeo Único/genética , Gravidez , Análise de Sequência de DNA , População BrancaRESUMO
Recent studies have indicated a role for caveolin in regulating cholesterol-dependent signaling events. In the present study we have analyzed the role of caveolins in intracellular cholesterol cycling using a dominant negative caveolin mutant. The mutant caveolin protein, cav-3(DGV), specifically associates with the membrane surrounding large lipid droplets. These structures contain neutral lipids, and are accessed by caveolin 1-3 upon overexpression. Fluorescence, electron, and video microscopy observations are consistent with formation of the membrane-enclosed lipid rich structures by maturation of subdomains of the ER. The caveolin mutant causes the intracellular accumulation of free cholesterol (FC) in late endosomes, a decrease in surface cholesterol and a decrease in cholesterol efflux and synthesis. The amphiphile U18666A acts synergistically with cav(DGV) to increase intracellular accumulation of FC. Incubation of cells with oleic acid induces a significant accumulation of full-length caveolins in the enlarged lipid droplets. We conclude that caveolin can associate with the membrane surrounding lipid droplets and is a key component involved in intracellular cholesterol balance and lipid transport in fibroblasts.
Assuntos
Caveolinas/metabolismo , Colesterol/metabolismo , Vesículas Citoplasmáticas/metabolismo , Metabolismo dos Lipídeos , Mutação/genética , Sequência de Aminoácidos , Androstenos/farmacologia , Animais , Anticorpos , Transporte Biológico/efeitos dos fármacos , Biomarcadores/análise , Brefeldina A/farmacologia , Caveolina 1 , Caveolinas/química , Caveolinas/genética , Linhagem Celular , Membrana Celular/química , Membrana Celular/metabolismo , Cricetinae , Vesículas Citoplasmáticas/química , Vesículas Citoplasmáticas/efeitos dos fármacos , Vesículas Citoplasmáticas/ultraestrutura , Retículo Endoplasmático/química , Retículo Endoplasmático/efeitos dos fármacos , Retículo Endoplasmático/metabolismo , Endossomos/química , Endossomos/metabolismo , Fibroblastos , Imunofluorescência , Genes Dominantes/genética , Homeostase , Microscopia Eletrônica , Microscopia de Vídeo , Ácido Oleico/farmacologia , Transdução de SinaisRESUMO
Cancer results from multiple genomic changes that affect DNA and its gene expression. The DNA sequences may be gained, lost or amplified, or translocated into different parts of the genome to form a fusion gene with oncogenic properties. The occurrence of specific chromosomal aberrations may be restricted to only one cancer type and it may be considered a primary carcinogenic event. Furthermore, the aberration profiles may be used to cluster tumors with similar origins. A variety of techniques exist for the detection of specific chromosomal and gene expression changes. However, the etiology of these molecular alterations remains unclear. Here we discuss the roles of Helicobacter pylori and asbestos burden as carcinogens that cause gastric cancer, mesothelioma and lung cancer.
Assuntos
Amianto/toxicidade , Carcinógenos/toxicidade , Helicobacter pylori/patogenicidade , Neoplasias/genética , Cromatina/genética , DNA/genética , Humanos , Neoplasias/diagnóstico , Neoplasias/microbiologiaRESUMO
Caveolae (small plasma membrane invaginations) and their coat proteins, caveolins, have attracted the attention of researchers in diverse fields, including cell biology, cardiovascular and cancer research. The tight association between caveolin and cholesterol governs the biochemical behaviour of caveolae and is emerging as an important characteristic in a number of processes assigned to these multifunctional organelles. In this review, selected aspects of the caveolin-cholesterol association and its potential functional implications are discussed.
Assuntos
Caveolinas/química , Caveolinas/metabolismo , Membrana Celular/química , Membrana Celular/metabolismo , Colesterol/metabolismo , Animais , Apolipoproteína A-I/metabolismo , Transporte Biológico , Humanos , Lipoproteínas HDL/metabolismoRESUMO
In this study, we compared the transport of newly synthesized cholesterol with that of influenza virus hemagglutinin (HA) from the endoplasmic reticulum to the plasma membrane. The arrival of cholesterol on the cell surface was monitored by cyclodextrin removal, and HA transport was monitored by surface trypsinization and endoglycosidase H digestion. We found that disassembly of the Golgi complex by brefeldin A treatment resulted in partial inhibition of cholesterol transport while completely blocking HA transport. Further, microtubule depolymerization by nocodazole inhibited cholesterol and HA transport to a similar extent. When the partitioning of cholesterol into lipid rafts was analyzed, we found that newly synthesized cholesterol began to associate with low-density detergent-resistant membranes rapidly after synthesis, before it was detectable on the cell surface, and its raft association increased further upon chasing. When cholesterol transport was blocked by using 15 degrees C incubation, the association of newly synthesized cholesterol with low-density detergent-insoluble membranes was decreased and cholesterol accumulated in a fraction with intermediate density. Our results provide evidence for the partial contribution of the Golgi complex to the transport of newly synthesized cholesterol to the cell surface and suggest that detergent-resistant membranes are involved in the process.