ABSTRACT
In this work, we report the synthesis of a new 1,3-thiazolium-5-thiolate derivative of a mesoionic compound (MIH 2.4Bl) and the characterization of its selective cytotoxicity on a panel of breast cancer cells lines. The cytotoxic effect of MIH 2.4Bl on breast cancer cell lines was determined by XTT and crystal violet assays, flow cytometry analysis, electron microscopy characterization, and terminal deoxynucleotidyl transferase (TdT) deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) apoptosis assays. As determined using XTT cell growth and survival assays, MIH 2.4Bl exhibited growth inhibition activity on most breast cancer cell lines tested, compared with normal human mammary epithelial cells. Three breast cancer cell lines (MCF-7, T-47D, and ZR-75-1) showed a more potent sensitivity index to growth inhibition by MIH 2.4Bl than the other breast cancer cell lines. Interestingly, these 3 cell lines were derived from tumors of Luminal A origin and have ER (estrogen receptor), PR (progesterone receptor), and HER2 (human epidermal growth factor receptor 2) positive expression. Additional analysis of cytotoxicity mediated by MIH 2.4Bl was performed using the MCF-7 cell line. MCF-7 cells displayed both time- and dose-dependent decreases in cell growth and survival, with a maximum cytotoxic effect observed at 72 and 96 hours. The MCF-7 cells were also characterized for cell cycle changes upon treatment with MIH 2.4Bl. Using flow cytometry analysis of cell cycle distribution, a treatment-dependent effect was observed; treatment of cells with MIH 2.4Bl increased the G2/M population to 34.2% compared with 0.1% in untreated (control) cells. Ultrastructural analysis of MFC-7 cells treated with MIH 2.4Bl at 2 different concentrations (37.5 and 75 µM) was performed by transmission electron microscopy. Cells treated with 37.5 µM MIH 2.4Bl showed morphologic changes beginning at 6 hours after treatment, while cells treated with 75 µM showed changes beginning at 3 hours after treatment. These changes were characterized by an alteration of nuclear morphology and mitochondrial degeneration consistent with apoptotic cell death. Results of a TUNEL assay performed on cells treated for 96 hours with MIH 2.4Bl supported the observation of apoptosis. Together, these results suggest that MIH 2.4Bl is a promising candidate for treating breast cancer and support further in vitro and in vivo investigation.
ABSTRACT
Agricultural pest control is a popular research topic, and essential oils are widely studied because they represent a promising alternative to synthetic insecticides. However, despite the increase in studies on pests, little work has been done on pesticide contamination of the predators feeding on insecticide-affected prey. Therefore, the objective of this study was to evaluate the effects of Lethal Dose 50 (LD50) from the essential oils of Mentha spicata L. and Melaleuca alternifolia Cheel on the histology, including histochemistry (from protein and carbohydrate analysis) and immunohistochemistry (from the evaluation of cellular apoptosis), of the midgut of fifth instar nymphs of Podisus nigrispinus (stinkbug) (Dallas). The periods of analysis were 12, 24, and 48 h after ingestion of Alabama argillacea Hübner caterpillars treated with the respective oils. The oil from M. spicata did not cause histological alterations or apoptosis in the insect. However, there was a reduction in the level of carbohydrates within the 48-h period. After 24 h, the oil of M. alternifolia caused an elongation of digestive cells and, after 48 h, cell lysis with the release of material into the lumen, suggesting tissue necrosis. The immunohistochemical study revealed no apoptotic process. There was a reduction in the neutral carbohydrate levels in the 24- and 48-h periods and in the number of regenerative cells, when compared to the control, after the period of 48 h. These results demonstrate that M. spicata oil has potential for use in cotton fields because it does not affect the vital characteristics of P. nigrispinus. However, the essential oil of M. alternifolia is not suitable for use as a pesticide because it is extremely toxic to predators.
