Determination of Rottlerin, a Natural Protein Kinases C Inhibitor, in Pancreatic Cancer Cells and Mouse Xenografts by RP-HPLC Method.
Rottlerin is a natural polyphenolic ketone isolated from the pericarps of Mallotus phillippinensis. In previous studies we showed that parenteral administration of rottlerin reduced tumor growth in murine xenograft models of pancreatic cancer. The aim of this study was to develop a simple and validated method for the quantitative determination of rottlerin in plasma and tumor tissues of mice fed a rottlerin diet. A xenograft model of pancreatic cancer was prepared by injection of 2×106HPAF-II cells subcutaneously into nude mice. One week before tumor implantation, mice were randomly allocated to standard diet (AIN76A) and standard diet supplement with 0.012% rottlerin (n=6 per group). Mice were sacrificed after 6 weeks on diets. Rottlerin was extracted from the plasma and tissues using protein precipitation-extraction and analyzed by reverse-phase HPLC-DAD method. The same HPLC method was also applied to determine rottlerin levels in conditioned culture media and in cell lysates from HPAF-II cells exposed to 25 µM concentration of rottlerin. A substantial amount of rottlerin was detected in tumor (2.11 ± 0.25 nmol/g tissue) and plasma (2.88 ± 0.41 µM) in mice fed rottlerin diet. In addition, significant levels of rottlerin (57.4 ± 5.4 nmol/mg protein) were detected in cell lysates from rottlerin-treated HPAF-II cells. These data indicate that rottlerin is efficiently absorbed in cells and tissues bothin vivoandin vitroand suggest a strong potential for rottlerin as a preventive or adjuvant supplement for pancreatic cancer.
Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
Qing-Yi
QY
Lu
Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
Lifeng
L
Zhang
Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA ; Veterans Affairs Greater Los Angeles Healthcare System, University of California, Los Angeles, CA, USA.
Aurelia
A
Lugea
Department of Surgery, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
Aune
A
Moro
Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA ; Veterans Affairs Greater Los Angeles Healthcare System, University of California, Los Angeles, CA, USA.
Mouad
M
Edderkaoui
Department of Surgery, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
Guido
G
Eibl
Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA ; Veterans Affairs Greater Los Angeles Healthcare System, University of California, Los Angeles, CA, USA.
Stephen J
SJ
Pandol
Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
Vay-Liang W
VL
Go
Cell uptake; HPLC; In vivo; Pancreatic cancer; Rottlerin; Tissue distribution