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Specific knock‐down of tissue non‐specific alkaline phosphatase mRNA levels inhibits intracellular lipid accumulation in 3T3‐L1 and HepG2 cells
Author(s) -
Chirambo George,
van Niekerk Chantal,
Crowther Nigel J.
Publication year - 2017
Publication title -
international journal of experimental pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.671
H-Index - 72
eISSN - 1365-2613
pISSN - 0959-9673
DOI - 10.1111/iep.12243
Subject(s) - intracellular , lipid droplet , alkaline phosphatase , adipocyte , transfection , messenger rna , chemistry , small interfering rna , 3t3 l1 , biochemistry , phosphatase , lipid metabolism , microbiology and biotechnology , biology , gene , enzyme , adipose tissue
Summary The use of non‐specific inhibitors of tissue non‐specific alkaline phosphatase ( TNSALP ) in pre‐adipocytes blocks intracellular lipid accumulation. TNSALP is also expressed in hepatocytes, which are known to accumulate lipid in a similar manner to pre‐adipocytes. The purpose of this study was to use specific silencing of TNSALP mRNA , using short interfering (si) RNA , to investigate the role of TNSALP in intracellular lipid accumulation in 3T3‐L1 and HepG2 cells. Cellular activity of TNSALP was measured using an automated colorimetric assay, and intracellular lipid accumulation was determined using the lipid‐specific dye, Oil Red O. Cells were transfected with si RNA directed against TNSALP mRNA , and expression of the TNSALP gene was determined at selected time points postinduction of lipid droplet formation. Expression of the TNSALP gene was inhibited by a maximum of 88 ± 1.9% ( P < 0.005 vs . control) 11 days after initiation of lipid droplet formation in the 3T3‐L1 cells and 80 ± 8.9% ( P < 0.05 vs . control) after 4 days in the HepG2 cells. This led to significant inhibition of both TNSALP activity and intracellular lipid accumulation in both cell lines. These data demonstrates that TNSALP plays an important role in the control of lipid droplet formation in both pre‐adipocyte and hepatocyte cell lines.