strychnine and Breast-Neoplasms

To examine the effect of brucine on the migration, invasion, adhesion and expressions of epithelial-to-mesenchymal transition (EMT) markers and matrix metalloproteinases (MMPs) in the highly metastatic breast cancer cell lines MDA-MB-231 and Hs578-T.. MDA-MB-231 and Hs578-T cells were divided to 4 groups: the control group (0.1% DMSO), and 25, 50 and 100 μmol/L brucine groups. The cell viability was determined using a CellTiter-Glo® luminescent cell viability. The scratch wound healing assay and tanswell migration assay were used to determine the migration ability of these cells treated by different concentrations of brucine. The proliferation rate, invasive potential and adhesive ability were respectively performed by colony formation assay, transwell invasion assay and adhension assay. The protein and mRNA expressions of EMT biomarkers, MMP-2 and MMP-9 were investigated by real-time reverse transcription polymerase chain reaction and Western blot.. Compared with the control group, brucine had little effect on cell viability or proliferation (P>0.05), but led to a dose-dependent decrease on migration, invasion, adhension of MDA-MB-231 and Hs578-T cells (P<0.01). Furthermore, brucine increased the protein and mRNA levels of EMT markers such as E-cadherin and β-catenin in MDA-MB-231 and Hs578-T cells, and decreased the protein and mRNA levels of mesenychmal markers such as vimentin and fibronectin, as well as the expressions of MMP-2 and MMP-9 (all P<0.01).. Brucine inhibited triple negative breast cancer cells metastasis potentially through EMT reversion and MMP-2 and MMP-9 inhibition.

Topics: Biomarkers, Tumor; Breast Neoplasms; Cell Adhesion; Cell Line, Tumor; Cell Movement; Cell Survival; Collagen; Drug Combinations; Epithelial-Mesenchymal Transition; Female; Humans; Laminin; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Neoplasm Invasiveness; Neoplasm Proteins; Proteoglycans; RNA, Messenger; Strychnine

To examine the effects of brucine on the invasion, migration and bone resorption of receptor activator of nuclear factor-kappa B ligand (RANKL)-induced osteoclastogenesis.. The osteoclastogenesis model was builded by co-culturing human breast tumor MDA-MB-231 and mouse RAW264.7 macrophages cells. RANKL (50 ng/mL) and macrophage-colony stimulating factor (50 ng/mL) were added to this system, followed by treatment with brucine (0.02, 0.04 and 0.08 mmol/L), or 10 μmol/L zoledronic acid as positive control. The migration and bone resorption were measured by transwell assay and in vitro bone resorption assay. The protein expressions of Jagged1 and Notch1 were investigated by Western blot. The expressions of transforming growth factor-β1 (TGF-β1), nuclear factor-kappa B (NF-κB) and Hes1 were determined by enzyme-linked immunosorbent assay.. Compared with the model group, brucine led to a dose-dependent decrease on migration of MDA-MB-231 cells, inhibited RANKL-induced osteoclastogenesis and bone resorption of RAW264.7 cells (P<0.01). Furthermore, brucine decreased the protein levels of Jagged1 and Notch1 in MDA-MB-231 cells and RAW264.7 cells co-cultured system as well as the expressions of TGF-β1, NF-κB and Hes1 (P<0.05 or P<0.01).. Brucine may inhibit osteoclastogenesis by suppressing Jagged1/Notch1 signaling pathways.

Topics: Animals; Bone Neoplasms; Breast Neoplasms; Cell Differentiation; Cells, Cultured; Female; Humans; Jagged-1 Protein; Macrophages; Mice; Osteoclasts; Receptor, Notch1; Signal Transduction; Strychnine

This study was designed to investigate the combination effects of brucine and gemcitabine, each with anticancer properties, in MCF-7 human breast cancer cells in culture. With regard to cell viability, effects of both the drugs and their combinations were inversely proportional to dose and time. For various proportional drug combinations studied, combination effects were analysed using CompuSyn software. The analyses revealed synergistic and/or additive effects regarding cell viability, anchorage-independent growth and cell migration. Combination analyses exhibited diversified impacts of the type of combination treatment, namely pretreatment with either drug followed by exposure to the other, or treatment with both drugs at the same time. Compared with untreated cells, combination treatment of asynchronised MCF-7 cells resulted in 17.2 × decrease in G2 phase, increasing G1 (2.1 × ) and S (1.5 × ) phase cells in cell cycle analysis. Brucine, either individually or in combination, but not gemcitabine, inhibited NF-kB subunit (p65) expression in MCF-7 cells.

Topics: Antimetabolites, Antineoplastic; Breast Neoplasms; Cell Cycle; Deoxycytidine; Drug Therapy, Combination; Female; Gemcitabine; Humans; MCF-7 Cells; Strychnine

With the health concerns of menopausal hormone replacement therapy, alternatives have been sought. Klimaktoplan® is a homeopathic formulation consisting of four main components and has been used for relief of menopausal symptoms for a long time. The study investigated the safety of Klimaktoplan® through its effect on the proliferation of breast cancer (MCF-7) and non-malignant mammary epithelial cells (MCF-10A).. MCF-7 and MCF-10A cells were cultured in 312.5, 625, and 1,250 μg/ml Klimaktoplan®. 17-Beta estradiol (E2) and medroxyprogesterone 17-acetate (MPA) were used for comparison with Klimaktoplan®. E2 only (0.001, 0.01, and 0.1 μM), and the combination of E2 (0.001, 0.01, and 0.1 μM) and MPA (0.01, 0.1, and 1 μM) were tested. Control cells for Klimaktoplan® and E2 groups were treated with dimethylsulfoxide (DMSO), and DMSO + ethanol was used for the combination group. Cellular proliferation was evaluated by the formation of insoluble formazan after incubation of 4 days.. Klimaktoplan® had a concentration-dependent anti-proliferative effect on breast cancer cells at 625 and 1,250 μg/ml, while not affecting proliferation of non-malignant mammary cells at any tested concentration. The effect of lactose was evaluated as lactose (the adjuvant of Klimaktoplan®) affect cell growth. E2 and lactose increased the proliferation of both malignant and non-malignant cells. The effect of E2 + MPA on the proliferation of malignant and non-malignant mammary cells was lower than estradiol only, but was higher than control.. Klimaktoplan® has an anti-proliferative effect on breast cancer cells, but not for non-malignant mammary epithelial cells, unlike E2 and E2 + P. With further research, KP would be a good alternative or additive in women with menopausal symptoms who wish to avoid conventional E or E + P hormone therapy.

Topics: Breast Neoplasms; Cell Proliferation; Cimicifuga; Epithelial Cells; Estradiol; Female; Homeopathy; Humans; MCF-7 Cells; Menopause; Phytotherapy; Plant Preparations; Progesterone; Sanguinaria; Strychnos

To study the effects of brucine on vascular endothelial growth factor (VEGF) expression and microvessel density (MVD) in a nude mouse model of bone metastasis due to breast cancer, and to assess the possible antitumor mechanism of brucine.. A syringe needle was used to directly inject 0.2 mL monoplast suspension (with 2×10(5) human breast cancer cells contained) into the bony femoral cortex of the right hind leg for modeling. Twenty-five nude mice were randomized into five groups and administered with an intraperitoneal injection of saline or drug for 8 consecutive days: model group (0.2 mL normal saline), low-dose brucine group (1.73 mg·kg(-1)), medium-dose brucine group (3.45 mg·kg(-1)), high-dose brucine group (6.90 mg·kg(-1)), and thalidomide group (200 mg·kg(-1)). Diet and activity were recorded, and the tumors were harvested 5 weeks later. The percentage of VEGF-positive cells was determined with hematoxylin and eosin staining and immunohistochemical staining, and MVD expression was determined by optical microscopy.. The VEGF expressions in brucine- or thalidomide-treated mice were significantly reduced as compared with mice in the model group (P <0.01). There were no significant difference between the high-dose brucine group and the thalidomide group (P >0.05). Significant difference was between the high- and low-dose brucine group P<0.05). Further, VEGF expression was significantly increased in the low- and medium-dose brucine groups compared with the thalidomide group (P <0.05). The MVD values in the three brucine and thalidomide groups were significantly lower than that in the model group (P <0.01). The MVD values in the medium- and high-dose brucine groups were not significantly different from those in the thalidomide group (P >0.05), while the MVD value showed a significant increase in the low-dose group compared with the thalidomide group (P <0.05).. Brucine could inhibit the growth of breast cancer to bone metastases, possibly by inhibiting tumor angiogenesis.

Topics: Animals; Bone Neoplasms; Breast Neoplasms; Cell Line, Tumor; Disease Models, Animal; Female; Humans; Immunohistochemistry; Mice; Mice, Inbred BALB C; Mice, Nude; Microvessels; Strychnine; Vascular Endothelial Growth Factor A; Xenograft Model Antitumor Assays

Isostrychnopentamine (ISP) is an indolomonoter-penic alkaloid that is present in the leaves of Strychnos usambarensis, an East African small tree. We have reported previously pro-apoptotic effects induced in vitro by ISP in the human HCT-116 colon cancer cell line, a model that displays relative sensitivity to apoptosis. In the present study, we observed that the in vitro growth inhibitory activities of ISP are similar in cancer cells that display sensitivity versus resistance to apoptosis. We made use of the U373 glioblastoma and the A549 non-small cell lung cancer (NSCLC) cell lines as models relatively resistant to apoptosis, and the human PC-3 prostate cancer cell line as a model relatively sensitive to apoptosis. While ISP induced transient decreases in [ATP]i and apoptosis in human U373 GBM cells, it did not provoke such features in A549 NSCLC cells. It thus seems that ISP-induced anti-cancer activity can lead to pro-apoptotic effects as a consequence, while apoptosis seems not to be the main cause by which ISP induces cancer cell death. ISP is a compound that merits further investigations in order to: i) identify the mechanism(s) of action by which it kills cancer cells, and ii) hemisynthesize novel ISP derivatives aiming to overcome, at least partly, the resistance of metastatic cancers to apoptosis.

Topics: Adenosine Triphosphate; Antineoplastic Agents, Phytogenic; Apoptosis; Brain Neoplasms; Breast Neoplasms; Carbolines; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; Drug Resistance, Neoplasm; Female; Glioblastoma; Humans; Inhibitory Concentration 50; Lung Neoplasms; Male; Neoplasms; Plant Leaves; Prostatic Neoplasms; Strychnos; Time Factors