Premium
Biochemical and NMR studies on structure and release conditions of RNA‐containing vesicles shed by human colon adenocarcinoma cells
Author(s) -
Ceccarini M.,
Guidoni L.,
Luciani A. M.,
Mariutti G.,
Rosi A.,
Viti V.
Publication year - 1989
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910440427
Subject(s) - vesicle , rna , adenocarcinoma , chemistry , biochemistry , pathology , biophysics , microbiology and biotechnology , cancer research , biology , medicine , membrane , cancer , gene
Large amounts of particulate material, mostly lipid vesicles, are released by human colon adenocarcinoma HCT‐8R cells when they are packed at high density in saline solution. RNA is also present in the released structures. Vesicle shedding is displayed only by healthy and viable cells. The process, in our experimental conditions, lasts up to 40 min, and can be restored by supplementing cells with nutrients and oxygen. RNA and lipids give rise to IH and 3IP NMR signals. The process is somehow related to a thermotropic transition observed by means of IH NMR spectroscopy for peculiar lipid domains in the plasma membrane. Analysis of 3IP NMR spectra of the phosphodiester groups, upon pH variation, indicates strong interaction between RNA and proteins in an assembled structure. A constant amount of polyA + RNA can be recovered from the vesicles. The electrophoretic pattern and in vitro protein synthesis indicate that mRNA can be isolated as a functionally active molecule with a major 5 Kb fraction.