© Georg Thieme Verlag KG Stuttgart · New York
In Vivo Antitumoral Activity of Stem Pineapple (Ananas comosus) Bromelain
Received: June 18, 2007
Revised: July 24, 2007
Accepted: August 27, 2007
24. September 2007 (online)
Stem bromelain (EC 188.8.131.52) is a major cysteine proteinase, isolated from pineapple (Ananas comosus) stem. Its main medicinal use is recognized as digestive, in vaccine formulation, antitumoral and skin debrider for the treatment of burns. To verify the identity of the principle in stem fractions responsible for the antitumoral effect, we isolated bromelain to probe its pharmacological effects. The isolated bromelain was obtained from stems of adult pineapple plants by buffered aqueous extraction and cationic chromatography. The homogeneity of bromelain was confirmed by reverse phase HPLC, SDS-PAGE and N-terminal sequencing. The in vivo antitumoral/antileukemic activity was evaluated using the following panel of tumor lines: P-388 leukemia, sarcoma (S-37), Ehrlich ascitic tumor (EAT), Lewis lung carcinoma (LLC), MB-F10 melanoma and ADC-755 mammary adenocarcinoma. Intraperitoneal administration of bromelain (1, 12.5, 25 mg/kg), began 24 h after tumor cell inoculation in experiments in which 5-fluorouracil (5-FU, 20 mg/kg) was used as positive control. The antitumoral activity was assessed by the survival increase (% survival index) following various treatments. With the exception of MB-F10 melanoma, all other tumor-bearing animals had a significantly increased survival index after bromelain treatment. The largest increase (∼318 %) was attained in mice bearing EAT ascites and receiving 12.5 mg/kg of bromelain. This antitumoral effect was superior to that of 5-FU, whose survival index was ∼263 %, relative to the untreated control. Bromelain significantly reduced the number of lung metastasis induced by LLC transplantation, as observed with 5-FU. The antitumoral activity of bromelain against S-37 and EAT, which are tumor models sensitive to immune system mediators, and the unchanged tumor progression in the metastatic model suggests that the antimetastatic action results from a mechanism independent of the primary antitumoral effect.
Bromeliaceae - Ananas comosus - cysteine proteinase - stem bromelain - antitumoral - antimetastatic
- 1 Rowan A, Buttle D. Pineapple cysteine endopeptidase. Methods Enzymol. 1994; 244 555-68.
- 2 Kelly G. Bromelain: a literature review and discussion of its therapeutic applications. Altern Med Rev. 1996; 1 405-10.
- 3 Braun J M, Schneider B, Beuth H J. Therapeutic use, efficiency and safety of the proteolytic pineapple enzyme Bromelain-POS in children with acute sinusitis in Germany. In Vivo. 2005; 19 417-21.
- 4 Lotti T. Controlled clinical studies of bromelain in the treatment of urogenital inflammation. Drugs. 1993; 46 144-6.
- 5 Mynott T, Ladhams A, Scarmato P, Engwerda C. Bromelain, from pineapple stems, proteolitically blocks activation of extracellular regulated kinase-2 in T cells. J Immunol. 1999; 163 9240-6.
- 6 Engwerda C, Andrew D, Ladhams A, Mynott T. Bromelain modulates T and B cell immune responses in vitro and in vivo . Cell Immunol. 2001; 210 66-75.
- 7 Hale L. Proteolytic activity and immunogenicity of oral bromelain within the gastrointestinal tract of mice. Int Immunopharmacol. 2004; 2 255-64.
- 8 Zavadová E, Desser L, Mohr T. Stimulation of reactive oxygen species production and cytotoxicity in human neutrophil in vitro after oral administration of a polyenzyme preparation. Cancer Biotechnol. 1995; 10 147-52.
- 9 Taussig S J, Szekerczes J, Batkin S. Inhibition of tumour growth in vitro by bromelain, an extract of the pineapple plant (Ananas comosus). Planta Med. 1985; 51 538-9.
- 10 Maurer H. Bromelain: biochemistry, pharmacology and medical use. Cell Mol Life Sci. 2001; 58 1234-45.
- 11 Kleef R, Delohery T, Boubjerg D. Selective modulation of cell adhesion molecules on lymphocytes by bromelain. Pathobiology. 1996; 64 339-46.
- 12 Engwerda C, Andrew D, Murphy M, Mynott T. Bromelain activates murine macrophages and natural killer cells in vitro . Cell Immunol. 2001; 210 5-10.
- 13 Kelly T. Fibroblast activation protein-alpha and dipeptidyl peptidase IV (CD26): Cell-surface proteases that activate cell signaling and are potential targets for cancer therapy. Drug Resist Updat. 2005; 8 51-8.
- 14 Gerard G. Anticancer treatment and bromelain. Agressologie. 1972; 4 261-74.
- 15 Lotz W. On the pharmacology of bromelain: an update with special regard to animal studies on dose dependent effects. Planta Med. 1990; 56 249-53.
- 16 Grabowska E, Eckert K, Fichtner I, Schulze-Forster K, Maurer H. Bromelain proteases suppress growth invasion and lung metastasis of B16F10 mouse melanoma cells. Int J Oncol. 1997; 11 243-8.
- 17 Batkin S, Taussig S, Szekerezes R. Modulation of pulmonary metastases (Lewis Lung carcinoma) by bromelain an extract of the pineapple stem (Ananas comosus). Cancer Invest. 1988; 6 241-2.
- 18 Hernández M, Chávez M, Márquez M, Rodríguez G, Santos R, González J. et al .Proceso de obtención de bromelina a partir de tallos de piña. Cuban Patent C12N 9/50 1997.
- 19 Anson M. The estimation of pepsin, trypsin, papain and cathepsin with hemoglobin. J Gen Physiol. 1938; 22 79-89.
- 20 Lowry O, Roserbrough N, Farr A, Randall R. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193 265-75.
- 21 Laemmli U. Cleavage of structural. Proteins during the assembly of the head bacteriophage T4. Nature. 1970; 237 680-5.
- 22 Leipner J, Iten F, Saller R. Therapy with proteolytic enzymes in rheumatic disorders. BioDrugs. 2001; 15 779-89.
- 23 Maurer H R, Hozumi M, Honma Y, Okabe-Kado J. Bromelain induces the differentiation of leukemic cells in vitro: an explanation for its cytostatic effects?. Planta Med. 1988; 54 377-81.
- 24 Tysnes B, Maurer H, Porwol T, Probst B, Bjerkvig R, Hoover F. Bromelain reversibly inhibits invasive properties of glioma cells. Neoplasia. 2001; 3 469-79.
- 25 Gaspani L, Limiroli E, Ferrario P, Bianchi M. In vivo and in vitro effects of bromelain on PGE (2) and SP concentrations in the inflammatory exudates in rats. Pharmacology. 2002; 65 83-6.
- 26 Folkman J. Role of angiogenesis in tumor growth and metastasis. Semin Oncol. 2002; 6 5-8.
- 27 Hale L P, Greer PK Sempowski G D. Bromelain treatment alters leukocyte expression of cell surface molecules involved in cellular adhesion and activation. Clin Immunol. 2002; 104 183-90.
- 28 Eckert K, Grabowska E, Stange R, Schneider U, Eschmann K, Maurer H R. Effects of oral bromelain administration on the impaired immunocytotoxicity of mononuclear cells from mammary tumor patients. Oncol Rep. 1999; 6 1191-9.
- 29 Beuth H J, Braun J M. Modulation of murine tumor growth and colonization by bromelain, an extract of the pineapple plant (Ananas comosus L.) In Vivo. 2005; 19 483-5.
- 30 Molloy C J, Pawlowski J E, Taylor D S, Turner C E, Weber H, Peluso M. Thrombin receptor activation elicits rapid protein tyrosine phosphorylation and stimulation of the raf-1/MAP kinase pathway preceding delayed mitogenesis in cultured rat aortic smooth muscle cells: evidence for an obligate autocrine mechanism promoting cell proliferation induced by G-protein-coupled receptor agonist. J. Clin Invest. 1996; 97 1173-83.
- 31 Gomes M TR, Mello V J, Rodrigues K C, Bemquerer M P, Lopes M TP, Faça V M. et al . Isolation of two plant proteinases in latex from Carica candamarcensis acting as mitogens for mammalian cells. Planta Med. 2005; 71 244-8.
Carlos E Salas, PhD
Departamento Bioquímica e Imunologia Instituto de Ciências Biológicas
Antonio Carlos 6627
Belo Horizonte 31270-901
eMail: [email protected]