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Analysis of L-type amino acid transporter in canine hepatocellular carcinoma.

Ogihara K, Naya Y, Sato R, Onda K, Ochiai H - J. Vet. Med. Sci. (2015)

Bottom Line: The inhibitor of LAT 2-aminobicyclo[2.2.1]heptane-2-carboxylic acid (BCH) reduced 90% of the activity at 1 mM.RT-PCR revealed distinct LAT1 expression compared with normal hepatocytes.These results indicated that the leucine transport activity of canine HCCs was due to LAT1.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Pathology, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara, Kanagawa 252-5201, Japan.

ABSTRACT
Analysis of L-type amino acid transport expression of hepatocellular carcinoma cells (HCCs) of the dog was performed. The leucine transport activity of canine HCCs was 0.628 ± 0.018 nmol/mg protein/min. The inhibitor of LAT 2-aminobicyclo[2.2.1]heptane-2-carboxylic acid (BCH) reduced 90% of the activity at 1 mM. The deduced amino acid sequences of canine LAT2, LAT3 and LAT4 were well conserved in mammalians, exhibiting 89, 88 and 77% homology, respectively. RT-PCR revealed distinct LAT1 expression compared with normal hepatocytes. Western blotting analysis confirmed the potent LAT1 expression in canine HCCs but not hepatocytes, and real-time RT-PCR analysis indicated that canine HCCs possessed 28 times higher LAT1 expression than hepatocytes. These results indicated that the leucine transport activity of canine HCCs was due to LAT1.

No MeSH data available.


Related in: MedlinePlus

Real-time RT-PCR analysis of mRNA of canine LAT1 in canine HCCs and normalhepatocytes (A) and Western blot analysis of LAT1 expression using antiserum againstthe C-terminus of canine LAT1 peptides (B).
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fig_008: Real-time RT-PCR analysis of mRNA of canine LAT1 in canine HCCs and normalhepatocytes (A) and Western blot analysis of LAT1 expression using antiserum againstthe C-terminus of canine LAT1 peptides (B).

Mentions: Amino acid sequences of canine LAT3 were compared with those of the human and mouse.Multiple sequence alignments were performed using the GENETYX program (ver. 10).Asterisks and dots indicate identical residues and conservative substitutions,respectively. The cDNA sequences data of canine LAT3 were deposited in the DNA DataBank of Japan (accession numbers AB924118).


Analysis of L-type amino acid transporter in canine hepatocellular carcinoma.

Ogihara K, Naya Y, Sato R, Onda K, Ochiai H - J. Vet. Med. Sci. (2015)

Real-time RT-PCR analysis of mRNA of canine LAT1 in canine HCCs and normalhepatocytes (A) and Western blot analysis of LAT1 expression using antiserum againstthe C-terminus of canine LAT1 peptides (B).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4478731&req=5

fig_008: Real-time RT-PCR analysis of mRNA of canine LAT1 in canine HCCs and normalhepatocytes (A) and Western blot analysis of LAT1 expression using antiserum againstthe C-terminus of canine LAT1 peptides (B).
Mentions: Amino acid sequences of canine LAT3 were compared with those of the human and mouse.Multiple sequence alignments were performed using the GENETYX program (ver. 10).Asterisks and dots indicate identical residues and conservative substitutions,respectively. The cDNA sequences data of canine LAT3 were deposited in the DNA DataBank of Japan (accession numbers AB924118).

Bottom Line: The inhibitor of LAT 2-aminobicyclo[2.2.1]heptane-2-carboxylic acid (BCH) reduced 90% of the activity at 1 mM.RT-PCR revealed distinct LAT1 expression compared with normal hepatocytes.These results indicated that the leucine transport activity of canine HCCs was due to LAT1.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Pathology, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara, Kanagawa 252-5201, Japan.

ABSTRACT
Analysis of L-type amino acid transport expression of hepatocellular carcinoma cells (HCCs) of the dog was performed. The leucine transport activity of canine HCCs was 0.628 ± 0.018 nmol/mg protein/min. The inhibitor of LAT 2-aminobicyclo[2.2.1]heptane-2-carboxylic acid (BCH) reduced 90% of the activity at 1 mM. The deduced amino acid sequences of canine LAT2, LAT3 and LAT4 were well conserved in mammalians, exhibiting 89, 88 and 77% homology, respectively. RT-PCR revealed distinct LAT1 expression compared with normal hepatocytes. Western blotting analysis confirmed the potent LAT1 expression in canine HCCs but not hepatocytes, and real-time RT-PCR analysis indicated that canine HCCs possessed 28 times higher LAT1 expression than hepatocytes. These results indicated that the leucine transport activity of canine HCCs was due to LAT1.

No MeSH data available.


Related in: MedlinePlus