[Changes of protein expression in HepG2 cells with CDK2 RNA interference].

Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology

PubMedID: 23643165

Shang J, Wang Z, Li J. [Changes of protein expression in HepG2 cells with CDK2 RNA interference]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2013;29(4):359-63.
OBJECTIVE
To investigate the effects of stable transfection of CDK2 siRNA on biological activities and nuclear proteins of human hepatocellular carcinoma HepG2 cells.

METHODS
HepG2 cells were transfected with the eukaryotic expression vector of P(Genesil-1-CDK2); via RNA interference and selected for the ones with stable transfection. We observed the changes in the cell growth curve and cell cycle. The mRNA contents of CDK2 and differentially expressed nucleoproteins were detected and analyzed by RT-PCR and two-dimensional (2D) electrophoresis-mass spectrum (MS)-database, respectively. Western blotting were used to confirm the differential protein expressions.

RESULTS
Compared with P(HK-siRNA);-HepG2 and untransfected groups, the proliferation of HepG2 cells in P(CDK2-siRNA);-HepG2 group was significantly inhibited (P<0.01), and the expression of CDK2 mRNA significantly decreased in P(CDK2-siRNA);-HepG2 group. Four proteins not expressed in P(CDK2-siRNA);-HepG2 cells were detected by 2D electrophoresis-MS, and they were further confirmed by Western blotting.

CONCLUSION
CDK2 siRNA significantly suppressed CDK2 mRNA expression and the proliferation of HepG2 cells, four proteins not expressed in p(CDK2-siRNA);-HepG2 cells are similar to ribosomal protein S12, ß-actin, zine finger 276 and chaperonin 10 related protein.