Determination of Rottlerin, a Natural Protein Kinases C Inhibitor, in Pancreatic Cancer Cells and Mouse Xenografts by RP-HPLC MethodQing-Yi Lu1*, Lifeng Zhang1, Aurelia Lugea1,2, Aune Moro3, Mouad Edderkaoui1,2, Guido Eibl3, Stephen J. Pandol1,2
and Vay-Liang W. Go1
- *Corresponding Author:
- Qing-Yi Lu
Center for Human Nutrition
Department of Medicine
University of California, Los Angeles
900 Veteran Ave., 14-165, Los Angeles
CA 90095, USA
E-mail: [email protected]
Received Date: December 14, 2012; Accepted Date: December 26, 2012; Published Date: Date: December 30, 2012
Citation: Lu QY, Zhang L, Lugea A, Moro A, Edderkaoui M, et al. (2012) Determination of Rottlerin, in Pancreatic Cancer Cells and Mouse Xenografts by RP-HPLC Method. J Chromat Separation Techniq 4:162. doi: 10.4172/2157-7064.1000162
Copyright: © 2012 Lu QY, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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 2×106 HPAF-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 both in vivo and in vitro and suggest a strong potential for rottlerin as a preventive or adjuvant supplement for pancreatic cancer.