Croton lechleri sap and isolated alkaloid taspine exhibit inhibition against human melanoma SK23 and colon cancer HT29 cell lines
Graphical abstract
Introduction
Croton lechleri Müll.Arg. (fam. Euphorbiaceae) is a traditional medicinal plant which produces a red sap, traditionally known as “Sangre de Drago” that is obtained carving the bark of plants having two–three or more years. Amazonic natives use this sap to treat several diseases as wound healing, gastrointestinal illness and also cancers (Jones, 2003). Furthermore, the dried sap is marketed in health products in Western countries. The phytochemical characterization of the sap has lead to the finding that the oligomeric proanthocyanidins and flavonols constitute almost the 90% of the dry weight (Cai et al., 1991). But various minor compounds also have been found; one is the alkaloid taspine found in the sap of mature trees (Perdue et al., 1979). Others are the lignan 3′,4-O-dimethylcedrusin (Cai et al., 1993a, Pieters et al., 1993), various diterpenoids (Cai et al., 1993b) and nor-isoprenoid blumenol derivatives (De Marino et al., 2008).
Proanthocyanidins and other condensed tannins are known to be protective against various pathologies, including cancer (Bobe et al., 2009, De Bruyne et al., 1999, Hanausek et al., 2011). Croton lechleri has been evaluated in cell lines such as KB, V-79, K562 and HeLa for anticancer activity showing low activity (Alfonso-Castro et al., 2012, Chen et al., 1994, Itokawa et al., 1991, Rossi et al., 2003, Vaisberg et al., 1989). This activity was mainly ascribed to the presence of the alkaloid taspine (Fig. 1) which has also been reported as the active compound with wound healing properties (Porras-Reyes et al., 1993, Vaisberg et al., 1989). Taspine is the main alkaloid which characterizes Croton lechleri sap, but it is also present in Leontice eversmanii Bge (Berberidaceae), Caulophyllum thalictroides (L.) Michx. and Calophyllum robustum Maxim. (Berberidaceae), Magnolia x soulangiana Soul.-Bod. (Magnoliaceae). Recently, interest is growing on the Croton lechleri sap and on its isolated alkaloid taspine (Fayad et al., 2009, Rollinger et al., 2006, Zhang et al., 2010).
The incidence rate of cutaneous malignant melanoma and colorectal cancer is increasing over the years and classical chemotherapy agents are still partly unsuccessful, therefore there exists a huge need of new therapeutic approaches against these cancers (Goffin et al., 2005). Further, malignant melanoma is the most serious of all skin cancers and strongly resistant to chemo- and radiotherapy. Epidemiological studies have shown that its incidence is rapidly increasing (Kruijff and Hoekstra, 2012).
In this framework of renewed researches, we have considered the activities of Croton lechleri and taspine on epithelial cancer in vitro also in relation with its traditional use, which is mainly for topical application. In this context, Croton lechleri sap and isolated taspine were studied in comparison with taxol and vinblastine, two natural anticancer agents, on the cell viability and the cell cycle, and on the α-tubulin structure of melanoma and colon cancer cell lines in culture.
Section snippets
Plant material
The red sap of Croton lechleri was collected by traditional way, carving the bark from trees growing in the province of Napo, Ecuador. The voucher code number (SdD 007) for the crude drug was deposited in the Department of Pharmaceutical and Pharmacological Sciences of Padua University.
The sap was freeze-dried in a lyophilizer and stored at −20 °C. For cell culture experiments, the freeze-dried SdD was solubilised in water.
Croton lechleri sap and taspine
Before lyophilisation, pH and density of the sap were determined. Dry
MTT assay
The pH of the sap from Croton lechleri was 3.9 and the density was 1.08 g/mL according with the literature data (Risco et al., 2003). The chemical characterization has been performed through the determination of the alkaloid taspine as previously reported (Froldi et al., 2009). To allow the conservation, we freeze-dried the fresh sap obtaining a crystal brown powder with a dry residue of 27.5±0.6%.
We studied the activity of Croton lechleri sap and taspine on cellular proliferation using MTT
Discussion
In this research, we studied the potential anticancer activity of Croton lechleri sap and its isolated alkaloid taspine in comparison with taxol and vinblastine on the growth of melanoma and colon cancer cell lines, studying cell proliferation, α-tubulin structure and cell-cycle. The sap (1–100 μg/mL) showed antiproliferative activity against all cell lines with a higher inhibition of SK23 cells growth, determined by MTT assay. Also taspine showed antiproliferative activity from 0.1 μg/mL against
Acknowledgements
This work was supported by the Italian Ministry for University and Research (MIUR) n. 60A04-5521/11 and 60A04-8432/12.
The authors acknowledge the traditional healers of the western Amazon basin of South America who use the latex of the Croton lechleri tree to treat many diseases. The authors also thank “Il Crisolito”s.r.l. (Spresiano, Treviso, Italy), Padri Giuseppini and Dr. Francesco Farnesi for providing the Croton lechleri sap from Ecuador.
References (35)
- et al.
Clerodane diterpenoids from Croton lechleri
Phytochemistry
(1993) - et al.
Diterpenes from Croton lechleri
Phytochemistry
(1993) - et al.
Polyphenolic compounds from Croton lechleri
Phytochemistry
(1991) - et al.
Activity of sap from Croton lechleri on rat vascular and gastric smooth muscles
Phytomedicine
(2009) - et al.
The current status of S-100B as a biomarker in melanoma
European Journal of Surgical Oncology
(2012) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays
Journal of Immunological Methods
(1983)- et al.
The ins and outs of tubulin acetylation: more than just a post-translational modification?
Cellular Signalling
(2011) - et al.
South American plants II: taspine isolation and anti-inflammatory activity
Journal of Pharmaceutical Sciences
(1979) - et al.
Evaluation of the mutagenic, antimutagenic and antiproliferative potential of Croton lechleri (Muell. Arg.) latex
Phytomedicine
(2003) - et al.
Sangre de grado Croton palanostigma induces apoptosis in human gastrointestinal cancer cells
Journal of Ethnopharmacology
(2002)
Activity of taspine isolated from Radix et Rhizoma Leonticis against estrogen-receptor-positive breast cancer
Fitoterapia
Discovery of novel taspine derivatives as antiangiogenic agents
Bioorganic & Medicinal Chemistry Letters
Taspine isolated from Radix et Rhizoma Leonticis inhibits growth of human umbilical vein endothelial cell (HUVEC) by inducing its apoptosis
Phytomedicine
Antitumor effect of Croton lechleri Mull. Arg. (Euphorbiaceae)
Journal of Ethnopharmacology
Stathmin activity influences sarcoma cell shape, motility, and metastatic potential
Molecular Biology of the Cell
Flavonoid consumption and oesophageal cancer among black and white men in the United States
International Journal of Cancer
Acetylated and detyrosinated alpha-tubulins are co-localized in stable microtubules in rat meningeal fibroblasts
Cell Motility Cytoskeleton
Cited by (32)
Natural Chemotherapeutic Agents for Cancer
2017, Multi-Scale Approaches in Drug Discovery: From Empirical Knowledge to In silico Experiments and BackEfficacy of Dragon's blood cream on wound healing: A randomized, double-blind, placebo-controlled clinical trial
2016, Journal of Traditional and Complementary MedicineCitation Excerpt :Dragon's blood has been used by different civilizations such as the Greeks, the Romans, and the Arabs.1 Croton lechleri has several medicinal properties, such as wound healing,1,2 cicatrizant,3 immunomodulator,1,4 analgesic, antiulcer, antidiarrheal,1 antibacterial,5 antiviral,6 antihemorrhagic,1 anti-inflammatory, antioxidant,1,3 mutagenic and antimutagenic,1,3,7 antitumor,1,8 anticancer,9 and cytotoxic effects.1,3 Proanthocyanidins are the main chemical constituent of the resin, >90% of the dry weight.10
Natural antitubulin agents: Importance of 3,4,5-trimethoxyphenyl fragment
2015, Bioorganic and Medicinal ChemistryMay EPH/Ephrin Targeting Revolutionize Lung Cancer Treatment?
2023, International Journal of Molecular Sciences