title: Prostaglandin A2 influences gene expression in an established insect cell line (BCIRL-HzAM1) cells.
abstract: Prostaglandins (PGs) and other eicosanoids are oxygenated metabolites of arachidonic acid and two other C(20) polyunsaturated fatty acids. While most well studied in mammals, PGs exert important actions in insects and virtually all other invertebrates. We have been researching the mechanisms of PG actions in established insect cell lines and reported earlier that two PGs, PGA(1) and PGE(1), influence gene and protein expression in HzAM1 cells. Here we report on further experiments with three 2-series PGs, PGA(2), PGE(2) and PGF(2α). In separate experiments we treated cells with each of the three PGs for 12 and 24h and then analyzed cell lysates by 2-D electrophoresis. Analysis of the gels by Delta2D software showed that PGA(2) influenced expression of 60 proteins while PGE(2) and PGF(2α) treatments led to expression changes for only a few proteins. All spots representing changes in protein expression were processed for analysis by MALDI TOF/TOF mass spectrometry. Bioinformatic analysis of the resulting sequences yielded in silico identifications of all proteins. The apparent changes in some proteins were confirmed by quantitative PCR, which demonstrated that changes in protein expression were parallel to changes in mRNA expression. We assorted the proteins into functional categories, including 1/cell structure and function; 2/cell protection and immunity; 3/energetics and metabolism; 4/nucleotide processing; 5/protein action and processing and 6/signal transduction. These findings substantially extend our idea that one mechanism of PG actions in insect cells is the modulation of gene and protein expression.
authors.affiliation: USDA/Agricultural Research Service, Biological Control of Insects Research Laboratory, Columbia, MO 65203, USA. stanleyd@missouri.edu
authors.forename: David W
authors.initials: DW
authors.lastname: Stanley
authors.forename: Cynthia
authors.initials: C
authors.lastname: Goodman
authors.forename: Shiheng
authors.initials: S
authors.lastname: An
authors.forename: Qisheng
authors.initials: Q
authors.lastname: Song
mesh_terms: D000818:Animals; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation / Q000187:drug effects; D019476:Insect Proteins / Q000096:biosynthesis* / Q000235:genetics; D007915:Lepidoptera / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D011454:Prostaglandins A / Q000494:pharmacology*; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D019476:Insect Proteins; D011454:Prostaglandins A; D012333:RNA, Messenger; C100008:prostaglandin A2