The electrical pulse application enhances intra-cellular localization and potentiates cytotoxicity of curcumin in breast cancer cells.

Breast cancer is the most common cancer of women, and fifth leading cause of mortality worldwide. Existing breast cancer regimens are costly and produce severe side effects. This highlights a need for the development of efficient novel therapies, which are cost effective and limit side effects. An electrical pulse (EP)-based chemo therapy, known as electrochemotherapy (ECT) using the natural compound curcumin could be an effective alternative. ECT is a non-surgical modality, which produces excellent anti-tumor efficacy at small drug concentrations due to increased uptake of drugs. In clinics, ECT is shown to be effective in treating advanced, recurrent, and metastatic breast cancers, which are refractory to multiple modalities. ECT with curcumin triggers apoptotic cell death in breast cancer cells and could be an effective alternative, due to curcumin's low cost and reduced side-effects. However, there is a lack of studies quantifying the uptake of curcumin in response to EP application. Towards this, we determined the uptake of different curcuminoids (curcumin, desmethoxycurcumin, and bisdemethoxycurcumin) upon EP application and their impact on cell cytotoxicity. Additionally, we studied the combined effect of calcium chloride (CaCl2) and a curcuminoids (Cur) mixture, based on initial studies suggesting calcium electroporation as a potential inexpensive anti-cancer treatment. Our results indicate EP with Cur increases cellular uptake, cell shrinkage, and cytotoxicity. The EP + Cur resulted in the highest uptake of the bisdemethoxycurcumin. Further, EP also potentiated the cytotoxicity of CaCl2 and of the Cur and CaCl2 combination against breast cancer cells and caused apoptosis. Our preliminary data pave the way to further studies on Cur and CaCl2 combination treating breast cancer.

Production of anthocyanins by Catharanthus roseus.

A stable cell suspension line of Catharanthus roseus producing anthocyanin was obtained. In this strain it was found that approximately 30% of cells regularly accumulated these metabolites and that anthocyanin accumulation occurred between the second half of log phase and the stationary phase of the culture growth cycle. The anthocyanins in the suspension cultures were compared with those biosynthetized in the flowers both of regenerated by somatic embryogenesis and field-grown plants. Six anthocyanins were identified in all the examined samples, three 3-O-glucosides and three 3-O-(6-O-p-coumaroyl) glucosides of petunidin, malvidin and hirsutidin. The hirsutidin coumaroyl glucoside has not been reported previously, and was predominat in all samples. The anthocyanin relative content was similar for cell suspensions and flowers from regenerated plants but different from field-grown plant flowers; instead, the total content was almost the same for the two flower types and higher compared to suspension culture content.

beta-Galactosidases with a lectin-like domain are expressed in strawberry.

Strawberry fruits (Fragaria x ananassa Duch.) undergo a marked softening during their ripening, and the process is accompanied by a release of free sugars with galactose among them. In this work total beta-galactosidase activity was measured in cell wall proteins from strawberry fruits at different developmental stages. Three full-length cDNAs (Fa beta gal1, Fa beta gal2 and Fa beta gal3, respectively) encoding different beta-galactosidases (EC 3.2.1.23) were isolated from a library representing red fruit transcripts. All of them could be detected both in fruits and in vegetative tissues. However, only Fa beta gal1 showed an increasing expression during the ripening stages up to a maximum in the red fruits, while the other two (Fa beta gal2 and Fa beta gal3) were mostly found in green fruits and became barely detectable during ripening proper. The three beta-galactosidase-encoding cDNAs were expressed in the yeast Pichia pastoris, and it was thus possible to demonstrate that each of them encode a beta-galactosidase. The expression of the three beta-galactosidase genes appears to be down-regulated by auxin, as already observed for other ripening-related genes of the non-climacteric strawberry. An unusual characteristic of two strawberry beta-galactosidases (Fa beta gal1 and Fa beta gal2) is that at the C-terminus of the enzymes a domain is found which is structurally related to known animal peptides with a sugar-binding ability.

Monitoring the production yields of vincristine and vinblastine in Catharanthus roseus from somatic embryogenesis. Semiquantitative determination by flow-injection electrospray ionization mass spectrometry.

A semiquantitative determination of two bis-indole antitumor alkaloids, vincristine and vinblastine, has been performed by flow-injection electrospray ionization mass spectrometric analysis of the extracts of Catharanthus roseus. Leaves and flowers of two different phenotypes (pink flower and white flower) obtained from somatic embryogenesis were thus examined and compared with the field-grown mother plant. Different amounts of vincristine and vinblastine were detected depending on the examined samples.