Overexpression of stearoyl-CoA desaturase-1 results in an increase of conjugated linoleic acid (CLA) and n-7 fatty acids in 293 cells.
The effect of human SCD1 heterologous expression on cellular fatty acid synthesis was investigated in the current study. The SCD1 gene expression cassette and PGK-neomycin-selectable marker cassette were co-introduced into HEK 293 cells by electroporation, and subsequently, SCD1 expression was evaluated by fatty acid analysis. RT-PCR analysis indicated that the foreign SCD1 gene could be expressed in transformed cell lines. Total lipid analysis of the transformed cells fed with vaccenic acid (t11-18:1) as a substrate showed that SCD1 expression resulted in an increase in c9t11-CLA from 0.73-1.03% to 2.69-2.86% (p<0.05) and that the conversion efficiency was elevated from 5.11-6.88% to 16.49-20.06% (p<0.05). Surprisingly, the concentration of t10c12-CLA was also increased, from 0.10-0.41% to 1.35-1.69% in SCD1 cells (p<0.05). SCD1 expression also resulted in a significant (p<0.05) increase in palmitoleic acid (16:1 n-7) from 1.56-2.26% to 3.47-4.04% and cis-vaccenic acid (18:1 n-7) from 2.42-3.97% to 6.20-7.22%, and the corresponding conversion ratio of n-7 fatty acid was elevated from 12.01-16.70% to 22.62-24.13% (p<0.05). This study demonstrates that the foreign SCD1 gene was expressed with high efficiency and induced elevated c9t11-CLA, t10c12-CLA, and n-7 fatty acid levels in mammalian cells.
Wu
Zhifang
Z
State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, PR China.
Li
Defa
D
Gou
Kemian
K
D002460:Cell Line; D005231:Fatty Acids, Unsaturated / Q000096:biosynthesis*; D006801:Humans; D044243:Linoleic Acids, Conjugated / Q000096:biosynthesis*; D013230:Stearoyl-CoA Desaturase / Q000096:biosynthesis* / Q000235:genetics
D005231:Fatty Acids, Unsaturated; D044243:Linoleic Acids, Conjugated; C523776:SCD1 protein, human; D013230:Stearoyl-CoA Desaturase