Pseudomonas Aeruginosa Preparations Of Anti-cholangiocarcinoma Role Of Experimental Research

Objective:Our experimental subjects consist of in-vitro cultured human cholangioca-rcinoma cells (QBC939) and nude mice carrying tumors by transplanted QBC939 cells. We observed the growth inhibition effect on both QBC9393 cells and those transplanted tumors following the treatment of pseudomonas aeruginosa (PA-MSHA). We, then, explored a practical and effective new method for adjuvant therapy for cholangiocarcinoma.Method:1. In vitro, cell viability and adhesion rate of human cholangiocarcinoma cells (QBC939) were measured. We use MTT assay to measure cell viability of QBC9393 at different PA-MSHA concentrations and treatment intervals, therefore determined the optimum PA-MSHA treatment effectiveness. MTT assay was used to test the proliferation inhibition rate of QBC939 cells with treatments of different PA-MSHA concentrations at different time. Cell morphology was observed under microscope and apoptosis was studied by DNA ladder agrose gel electrophoresis. QBC9393 cell cycle was also analyzed using flow cytometry under PA-MSHA treatment.2. We transplanted the tumor QBC939 cells into nude mice and compared the PA-MSHA treated and untreated subjects. We calculated tumor formation rate, tumor growth inhibition rate and studied histomorphology & pathomorphism on heart, liver, spleen, lung, kidney, and pancreas in these nude mice.Result:MTT results suggest that QBC939 cell growth inhibition rates by PA-MSHA are time and dose-dependent in vitro. Through DNA ladder and flow cytometry assays, we found that PA-MSHA can induce cell apoptosis and G₂/M cell cycle phase arrest. We next examined the effect of PA-MSHA on QBC939 transplanted nude mice, and found that all the superficial lymph nodes and abdominal lymph node have no sign of metastasis. Peritoneal metastasis and ascites are also not seen in the nude mice. The size and weight of PA-MSHA treated tumor was significantly smaller than the non-treated one. HE staining showed that tumor tissue shares similar characteristics with primary cell line. However, treated tumor has lower density and more nest necrosis. No metastasis was found in heart, liver, spleen, lung, kidney and pancreas of the nude mice.Conclusions:1. PA-MSHA clearly inhibits the growth of human cholangiocarcinoma QBC939 cells at the concentration of 2-6x10⁸ cfu/ml in vitro. 2. PA-MSHA can induce apoptosis and cell cycle arrest of the QBC939 cells.3. The QBC939 tumor in the nude mice has shrunk after PA-MSHA treatment. 4. Tumor treated with PA-MSHA has lower density and more nest necrosis. 5. PA-MSHA has no obvious side effects on heart, liver, spleen, lung, kidney and pancreas in cancer bearing nude mice. 6. PA-MSHA might be a promising anti-cancer drug, particularly in the local treatment. These observations encourage further study of this drug.