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• W2313305767 abstract "B22 Pancreatic cancer is the fifth leading cause of cancer mortality in females and fourth in males in the United States. Diagnosed later than other cancers, there is poor prognosis and incidence and mortality has not experienced many changes. Approximately 30,000 new cases are diagnosed each year, and the incidence is highest in African Americans. Less than 5% of patients survive more than 5 years after an initial diagnosis. Natural sources are continually been explored and several novel compounds including Doxorubicin (Adriamycin) produced by a member of the genus Streptomyces, and Paclitaxel (Taxol®) originally derived from plant source are presently employed in therapy. However, members of the genus Bacillus, already established producers of antimicrobial compounds, have not received tremendous investigation regarding their production of antitumor compounds. In response to this we investigated the induction of apoptosis in MIA PaCa-2 pancreatic carcinoma cells, treated with varying concentrations of the extract fermented by a novel Bacillus subspecies 14135. In previous experiments we have demonstrated the filtrate’s antineoplastic ability employing MCF7 breast adenocarcinoma, NCI-H526 lung carcinoma and LNCaP-clone FGC prostate carcinoma cells. 25cm3 flask cultured MIA PaCa-2 cells were plated with 5.0 x 104 cells/ml in 96-well cell culture plates, incubated for 72hrs, at 37oC, 5% CO2, andunder controlled humidity. At approx 95% confluence selected wells were treated with definite quantities of the extract containing precise concentration of native protein. In earlier studies employing GC-MS fractionation we identified Taxol® (Paclitaxel) treated and untreated tumor cells, along with vehicle treated and non-tumor treated reference line cells were employed as controls. Following incubation at 37oC, for 24hr to 48hrs, cells were then checked to confirm cytotoxicity and apoptosis. These were assessed through microscopic observations, employing trypan blue dye exclusion analysis, CellTiter-Blue® cell viability assay, and Annexin V- FITC® apoptosis assay. The latter is an indicator of plasma membrane destruction, which was revealed following fluorescence microscopy. In earlier studies where we employed GC-MS fractionation, several compounds were identified including Pyrollo [1, 2-a] pyrazine-1, 4-dione and Pyrollo [1, 2-a] piperazine-3, 6-dione. We feel that these compounds may be involvd in the cytotoxic activity. Results were conclusive regarding the demonstration of cytotoxicity, as well as very strong evidence indicative of associated cellular shrinkage. Untreated, vehicle treated and extract treated non-tumor reference line cells remained unaffected. These results indicate the presence of compounds, which appear to selectively induce apoptosis in PaCa-2 tumor cells, and could greatly impact future cancer research." @default.
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• W2313305767 date "2007-11-01" @default.
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• W2313305767 title "Apoptosis induced in PaCa-2 cancer cells with novel Bacillus fermented extract" @default.
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