Effects Of Pseudolaric Acid B On The Proliferation And Apoptosis Of Human Pancreatic Carcionma PANC-1Cell Lines

Objective：To detect the effects of PAB on the proliferation and apoptosis of PANC-1celllines. Investigate the probable mechanisms of suppresses and apoptosis and provide a basistheory for further clinical research and application.Methods: Test the effect of PAB on inhibiting proliferation of PANC-1cell lines by CCK-8test. The morphology of apoptosis of PANC-1cell lines was examined by fluorescencemicroscope after hoechst33258staining, while the clone formation of PANC-1cell lineswas detected by clone formation assay. The cell cycle and the percentage of hypodiploidcells of PANC-1cell lines treated with PAB for48h were determined with propidiumiodide (PI) staining by flow cytometry. Apoptosis was detected with Annexin v-pi dualparameter by flow cytometry. Western blot technique was used to detect apoptosis relatedproteins caspase-3, caspase-9and JNK expression levels.Results: PAB inhibited the growth of PANC-1cell lines in a time-and dose-dependentmanner. When the concentration is higher, the inhibitory rate is higher. Compared withcontrol group, the clone formation of PANC-1cell lines treated with PAB for7days wasalso significantly inhibited. After treated with PAB for48h, apoptosis morpholoficalalterations, such as cell number reduced obviously, some cells cytoplasmic reduce,chromation sondensation and apoptotic bodies were observed by staining with Hoechst33258under the fluorescence microscope. Compared with control group, After the cellswere treated with different concentration of PAB for48h,the ratio of G0/G1phasedecreased, ratio of G2/M phase increased obviously. The western blot results showed thatthe PAB can improve the caspase-3, caspase-9and JNK protein expression. And high PABconcentration could rise the protein expression levels. Conclusion:1. PAB could inhibite the proliferation of Human Pancreatic Carcionma PANC-1cell linesand have dose-dependent and time-dependent.2. The numbers of clone of PANC-1cells can be inhibited by PAB after7days effectively.3. PAB could induced apoptosis of Human Pancreatic Carcionma PANC-1cell lines. PABcould reduce cell number and cytoplasm, nucleus condensation and induce apoptoticmorphology change by Hoechest33258staining.4. PAB could inhibit cell proliferation at the G2/M phase.5. PAB can induced apoptosis, and the mechanism maybe related with activation ofcaspase-3, caspase-9and the JNK signaling pathway.