Alkaloids-rich fraction of Crotalaria sessiliflora induces cell growth arrest and apoptosis by up-regulating cyclin B1 signaling in pancreatic cancer cells

【作者】 曾军英； 李胜华； 伍贤进；

【Author】 Jun-ying Zeng;Sheng-hua Li;Xian-jin WU;The Department of Life Science,Huaihua University;Key Laboratory of Hunan Province for Study and Utilization of Ethnic Medicinal Plant Resources;Key Laboratory of Hunan Higher Education for Hunan-Western Medicinal Plant and Ethnobotany;

【机构】 民族药用植物资源研究与利用湖南重点实验室； 湘西药用植物与民族植物学湖南省高校重点实验室； 怀化学院生命科学系；

【摘要】 ETHNOPHARMACOLOGICAL RELEVANCE:A medicinal herb Crotalaria Sessiliflora has been used to treat various diseases including diuretic,cardiotonic,analgesic,and anticancer in folk traditional medicine in Southern of China.The aim of the present study was to investigate the anticancer activity of Crotalaria Sessiliflora in pancreatic cancer cells.MATERIALS AND METHODS:The cytotoxic and apoptotic effect of alkaloids-rich fraction of Crotalaria sessiliflora(ACS) on human pancreatic cancer cells 13.6pl and Bxpc-3 cells were evaluated using MTT and TUNEL assays,and Annexin V/Propidium iodide detection by flow cytometry.The effect of ACS on cell cycle was analyzed by flow cytometry with propidium iodide.The apoptotic and cell cycle effects were also evaluated by RT-PCR and western blot analysis.RESULTS:Alkaloids-rich fraction of Crotalaria sessiliflora repressed clonogenecity and proliferation,induced apoptosis,and enhanced accumulation in the G2 phase of pancreatic cancer cells.In addition,a second population of cells expressing cyclin Bl had developed polyploidy and DNA content consistent with 8 N chromosomes.Compared with adriamycin,a known inducer of p53,ACS-treated pancreatic cancer cells expressed p53 only at high concentrations and p53 expression was not coordinated with G2 arrest or polyploid.Furthermore,ACS suppressed the transcription of genes associated with cell cycle regulation,anti-apoptosis,and invasion.CONCLUSION:These data showed that ACS blocks proliferation and induces apoptosis in human pancreatic cancer cells through G2 arrest in association with persistent cyclin Bl expression,failure to completely activate cdc2,and continued DNA synthesis through a pathway that is unrelated to altered expression of p53.ACS can be used as an anticancer drug for pancreatic carcinoma patients.更多还原

【Abstract】 ETHNOPHARMACOLOGICAL RELEVANCE:A medicinal herb Crotalaria Sessiliflora has been used to treat various diseases including diuretic,cardiotonic,analgesic,and anticancer in folk traditional medicine in Southern of China.The aim of the present study was to investigate the anticancer activity of Crotalaria Sessiliflora in pancreatic cancer cells.MATERIALS AND METHODS:The cytotoxic and apoptotic effect of alkaloids-rich fraction of Crotalaria sessiliflora(ACS) on human pancreatic cancer cells 13.6pl and Bxpc-3 cells were evaluated using MTT and TUNEL assays,and Annexin V/Propidium iodide detection by flow cytometry.The effect of ACS on cell cycle was analyzed by flow cytometry with propidium iodide.The apoptotic and cell cycle effects were also evaluated by RT-PCR and western blot analysis.RESULTS:Alkaloids-rich fraction of Crotalaria sessiliflora repressed clonogenecity and proliferation,induced apoptosis,and enhanced accumulation in the G2 phase of pancreatic cancer cells.In addition,a second population of cells expressing cyclin Bl had developed polyploidy and DNA content consistent with 8 N chromosomes.Compared with adriamycin,a known inducer of p53,ACS-treated pancreatic cancer cells expressed p53 only at high concentrations and p53 expression was not coordinated with G2 arrest or polyploid.Furthermore,ACS suppressed the transcription of genes associated with cell cycle regulation,anti-apoptosis,and invasion.CONCLUSION:These data showed that ACS blocks proliferation and induces apoptosis in human pancreatic cancer cells through G2 arrest in association with persistent cyclin Bl expression,failure to completely activate cdc2,and continued DNA synthesis through a pathway that is unrelated to altered expression of p53.ACS can be used as an anticancer drug for pancreatic carcinoma patients.更多还原