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Determination of Rottlerin, a Natural Protein Kinases C Inhibitor, in Pancreatic Cancer Cells and Mouse Xenografts by
Qing-Yi Lu1, Lifeng Zhang1, Aurelia Lugea2
1Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
Abstract:
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.
Insights
Rottlerin, a natural compound, was effectively quantified in mouse plasma and tumors. This study demonstrates rottlerin
Area of Science:
- Pharmacology and Toxicology
- Natural Product Chemistry
- Cancer Research
Background:
- Rottlerin, a polyphenolic ketone from Mallotus phillippinensis, has shown potential in reducing pancreatic cancer growth.
- Previous research indicated rottlerin's efficacy via parenteral administration in murine models.
Purpose of the Study:
- To develop and validate a quantitative method for determining rottlerin levels in mouse plasma and tumor tissues.
- To assess rottlerin absorption and distribution in vivo and in vitro.
Main Methods:
- A xenograft model of pancreatic cancer was established using HPAF-II cells in nude mice.
- Mice were fed a standard diet or a diet supplemented with 0.012% rottlerin for six weeks.
- Rottlerin was extracted and quantified using reverse-phase HPLC-DAD in plasma, tumor tissues, cell lysates, and culture media.
Main Results:
- Significant levels of rottlerin were detected in tumor tissues (2.11 ± 0.25 nmol/g) and plasma (2.88 ± 0.41 µM) of mice on the rottlerin diet.
- High concentrations of rottlerin (57.4 ± 5.4 nmol/mg protein) were found in HPAF-II cell lysates after in vitro exposure.
- The HPLC-DAD method proved effective for rottlerin quantification.
Conclusions:
- Rottlerin is efficiently absorbed in both cellular and tissue matrices, in vivo and in vitro.
- These findings support rottlerin's potential as a preventive or adjuvant therapeutic agent for pancreatic cancer.
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