Wang Tao,Lei Zengjie,Wang Bin,Li Qing,Wang Chunhua,Xiong Ji,Lai Shujie,Liu Qin,Chen Dongfeng.Variation of ZCWCC1 protein expression in steatosis of L02 hepatocytes[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(02):89-94.
Wang Tao,Lei Zengjie,Wang Bin,Li Qing,Wang Chunhua,Xiong Ji,Lai Shujie,Liu Qin,Chen Dongfeng.Variation of ZCWCC1 protein expression in steatosis of L02 hepatocytes[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(02):89-94.DOI:
Variation of ZCWCC1 protein expression in steatosis of L02 hepatocytes
Objective To determine the expression profile of ZCWCC1 (Zinc-finger CW-type coiled-coil protein 1) protein in the process of steatosis of immortalized hepatocyte cell line L02 (hereinafter referred to as L02 cells). Methods L02 cells were cultured in the medium containing 500 μmol/L oleic acid to induce steatosis
then confirmed by oil red O staining. Immunofluorescence assay and fluorescent confocal microscopy were employed to observe the protein levels of ZCWCC1 during induction of steatosis
and Western blotting was used to semi-quantify the results. Lysine protease inhibitor
N-Ethylmaleimide (NEM)
and proteasome inhibitor
MG132 were used to enhance or block the ubiquitin-proteasome pathway which is important for protein degradation. Results Oil red O staining and cellular triglyceride measurement confirmed the establishment of L02 steatosis model. Immunofluorescence assay and Western blot analysis indicated that ZCWCC1 was mainly located in the nucleus
and its expression level was descending during the induction of steatosis (56 283±303
56 753±539
55 561±996
56 257±326
48 339±712
and 43 022±2 198 respectively in 0
3
6
12
24 and 48 h after induction
P0.01). NEM treatment resulted in enhanced degradation of ZCWCC1 in the L02 cells
while MG132 inversed the descending tendency of ZCWCC1 during L02 steatosis. Conclusion Protein level of ZCWCC1 was descending in the process of steatosis of hepatocyte L02 cells
which might be relative to the ubiquitin-proteasome pathway.