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Inhibition of HLA‐G Production in JEG‐3 Choriocarcinoma Cells by RNA Interference
Author(s) -
Wengenmayer Tobias,
Poehlmann Tobias G.,
Markert Udo R.
Publication year - 2004
Publication title -
american journal of reproductive immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.071
H-Index - 97
eISSN - 1600-0897
pISSN - 1046-7408
DOI - 10.1111/j.1600-0897.2004.00150.x
Subject(s) - rna interference , human leukocyte antigen , blot , choriocarcinoma , biology , trophoblast , cell culture , immune system , hla g , rna , oligonucleotide , microbiology and biotechnology , antigen , cancer research , immunology , gene , fetus , genetics , pregnancy , placenta
Human leukocyte antigen‐G (HLA‐G) plays a major role in escape of trophoblast cells from maternal cytotoxicity. Unless its malignant transformation, Jeg‐3 choriocarcinoma cell line maintained the capacity of HLA‐G production. For the analysis of function and mechanisms of HLA‐G‐induced immune regulation, a human cellular model with suppressed HLA‐G would be very helpful. RNA interference (RNAi) is a very elegant method for this approach, but the design of appropriate oligonucleotide sequences may provide difficulties. We designed oligonucleotides to interfere exclusively with HLA‐G mRNA, which were applied at different concentrations to 50% confluent Jeg‐3 cells. After 36 hr, the HLA‐G content in Jeg‐3 cells was analyzed by Western blots. Applying the described RNAi method and oligonucleotides the cellular content of HLA‐G was dose‐dependently reduced as assessed in several independent Western blots. This method provides a tool for extensive investigation of HLA‐G functions in vitro and in vivo .