Functional consequences of knocking down porcine prostaglandin synthases in SK-6 swine kidney cell line.
We hypothesized that in vitro knock-down of the previously cloned genes of prostaglandin synthases will result in a reduction of synthesis, and thus secretion of their respective products, i.e., prostaglandin (PG) E2 or PGF2α. For this purpose, we designed short hairpin RNA (shRNA)-encoding constructs to knock down porcine mPGES-1 and PGFS (also named as AKR1CL1 in GenBank) and used them to transfect swine kidney SK-6 cells. Knocking down PGFS or mPGES-1 transcripts resulted in at least 50% inhibition of protein expression of each respective enzyme, as well as a reduction in the production of their respective prostaglandin, PGF(2α) or PGE(2). These results confirmed the identities of PGFS and mPGES-1. Moreover, they illustrate a unique opportunity to use the gene knock-down constructs in primary endometrial cells in order to study their biological roles in the porcine endometrium, particularly during the establishment of pregnancy.
Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland. Electronic address: waclawik@pan.olsztyn.pl.
Agnieszka
A
Waclawik
Department of Physiology, University of Turku, 20520 Turku, Finland; Faculty of Natural Sciences and Technology, Åbo Akademi University, Turku, Finland.
Adolfo
A
Rivero-Müller
Department of Physiology, University of Turku, 20520 Turku, Finland.
Nafis A
NA
Rahman
Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland.
Adam J
AJ
Ziecik
D000818:Animals; D002460:Cell Line; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D055785:Gene Knockdown Techniques; D007668:Kidney / Q000166:cytology*; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics* / Q000378:metabolism; D013552:Swine
D011451:Prostaglandin-Endoperoxide Synthases
Gene knock-down; Pig; Prostaglandins; SK-6 cells; ShRNA