cyclin-d1 and fengycin

To explore the potential clinical anti-tumor roles of Bacillus subtilis fmbJ-derived fengycin on cell growth and apoptosis in colon cancer HT29 cell line.Fengycin was extracted from Bacillus subtilis fmbJ and detected using HPLC. The effects of different concentration of fengycin on colon cell HT29 cell activity at different time points were analyzed using MTT assay. ROS level in colon HT29 cells affected by fengycin was detected using DCFH-DA method, followed by measuring the effects of fengycin on HT29 cell apoptosis and cell cycle by flow cytometry. The effects of fengycin on Bax/Bcl-2, CDK4/cyclin D1, Caspase-6 and Caspase-3 expressions in HT29 cells were analyzed using western blot. Also, mRNA levels of Bax/Bcl-2 and CDK4/cyclin D1 in HT29 cells affected by fengycin were analyzed using qRT-PCR.Compared with controlss, 20 μg/mL of fengycin performed an inhibit role on HT29 cell growth of at 3 day (P<0.05), and high dose of fengycin showed more excellent effect on inhibiting HT29 cell growth with time increasing. Besides, fengycin could induce HT29 cell apoptosis and affect the cell cycle arrest at G1. ROS level in HT29 cells treated by fengycin was significantly increased compared with that in control group (P<0.05). Western blot analysis showed that after being treated with fengycin, Bax, Caspase-3, and Caspase-6 expressions were increased, however, Bcl-2, and CDK4/cyclin D1 expressions were decreased (P<0.05).Our study suggested that fengycin may play certain inhibit roles in the development and progression of colon cancer through involving in the cell apoptosis and cell cycle processes by targeting the Bax/Bcl-2 pathway.

Topics: Antineoplastic Agents; Apoptosis; Bacillus subtilis; bcl-2-Associated X Protein; Caspase 3; Caspase 6; Cell Line, Tumor; Cell Proliferation; Colonic Neoplasms; Cyclin D1; Cyclin-Dependent Kinase 4; G1 Phase Cell Cycle Checkpoints; HT29 Cells; Humans; Lipopeptides; Reactive Oxygen Species

To investigate the antitumor activity and action mechanism of fengycin using the human lung cancer cell line 95D. The antitumor activity of fengycin was tested in vitro and in vivo. Reactive oxygen species production, Ca(2+) uptake, and mitochondrial membrane potential loss induced by fengycin in 95D cells were measured by flow cytometry and a laser confocal microscope. Lactate dehydrogenase release and caspase activity in fengycin-treated 95D cells were assayed using cytotoxicity detection kits. Apoptosis triggered by fengycin was identified by 4,6-diamidino-2-phenylindole (DAPI) staining and flow cytometry. The effects of fengycin on cell-cycle and apoptosis-related proteins were evaluated by quantitative reverse-transcription PCR and western blot. Treatment with fengycin not only significantly decreased cell proliferation in various cancer cell lines including 95D but inhibited the growth of xenografted 95D cells in nude mice. Fengycin also induced reactive oxygen species production and Ca(2+) uptake, as well as lactate dehydrogenase release and mitochondrial membrane potential loss. Further experiments showed that fengycin could trigger apoptosis in 95D cells and cause cell-cycle arrest at the G0/G1 stage by downregulating cyclin D1 and cyclin-dependent kinase 4 (CDK4). While investigating caspase activity and the expression of apoptosis-related proteins, fengycin was found to induce apoptosis in 95D cells through the mitochondrial pathway, evidenced by increased caspase activity, Bax expression, and cytochrome c release into the cytoplasm, as well as decreased Bcl-2 levels. Fengycin can inhibit the growth of the cancer cell line 95D by regulating the cell cycle and promoting apoptosis, suggesting that it may have potential as an anticancer treatment.

Topics: Antineoplastic Agents; Apoptosis; Calcium; Caspases; Cell Cycle; Cell Line, Tumor; Cyclin D1; Cyclin-Dependent Kinase 4; Flow Cytometry; Gene Expression Regulation, Neoplastic; Genes, Neoplasm; Humans; L-Lactate Dehydrogenase; Lipopeptides; Lung Neoplasms; Microscopy, Confocal; Mitochondrial Membranes; Reactive Oxygen Species