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Updated: Jul 15, 2026
DNA Vector-based RNA Interference to Study Gene Function in Cancer
Published on: June 4, 2012
[Construction of eukaryotic expression plasmid expressing siRNA targeting HLA-G gene and detection of its specific
Jian-hui Chen1, Yuan-qing Yao, Kai-bo Hou
1Department of Obstetrics and Gynecology, Tangdu Hospital, Xi'an 710038, China. jhch0904@163.com
Aim:
To investigate the effect of HLA-G siRNA on the protection of trophoblast cells from NK cell lysis, an HLA-G-targeting siRNA (small interfering)-expressing plasmid was constructed.
Methods:
HLA-G siRNA plasmid, pSuppressor-U6-neo-HLA-G, was constructed and transfected to JEG-3 cells. The level of messenger RNA and protein of HLA-G in the transfected cells was detected by RT-PCR and Western blot. NK-92MI cells were co-cultured with pSuppressor-U6-neo-HLA-G transfected JEG-3 cells. The cytotoxicity of NK-92MI cells against the transfected cells was analyzed by LDH releasing assay.
Results:
Downregulation of HLA-G in HLA-G siRNA plasmid transfected JEG-3 cells was confirmed by RT-PCR and Western blot. Compared with JEG-3 cells without HLA-G without HLA-G siRNA plasmid transfection, the transfected JEG-3 cells showed higher lytic activity.
Conclusion:
HLA-G siRNA can increasing NK cytolysis against JEG-3 cells. HLA-G plays a role in preventing of trophoblast cells from NK cytotoxicity.
