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1、齐墩果酸对肝癌细胞Hep3B增殖和凋亡的作用研究黄开顺1,朱链链2,刘丹2,邹黎2,雷琪2,葛蹼2,陈志琼1,2*(重庆医科大学1.生命科学研究院;2.药学院药物分析教研室;3.基础医学院病理生理教研室,重庆400016)摘要目的研究齐墩果酸对人肝癌细胞株Hep3B增殖的影响及其凋亡与细胞内钙离子浓度([Ca2+]i)关系。方法不同浓度齐墩果酸作用于肝癌Hep3B细胞以后,DAPI染色,以荧光显微镜观察细胞形态变化;用四甲基偶氮唑蓝(MTT)法检测Hep3B细胞增殖情况;流式细胞仪检测细胞周期改变、细胞凋亡率和细胞内钙离子浓度;对钙离子荧光强度与细胞凋亡率
2、之间进行相关性分析。结果细胞凋亡发生;不同浓度齐墩果酸能够抑制Hep3B细胞株增殖,且在5~100μg/mL范围内呈剂量依赖性,药物作用细胞24、48h的IC50分别为86.04、93.29μg/mL;各处理组细胞周期在G2/M期产生阻滞、([Ca2+]i)较对照组显著增加,细胞凋亡率和[Ca2+]i增加与药物浓度存依赖关系;钙离子荧光强度与细胞凋亡率之间存在线形相关性。结论齐墩果酸能够抑制肝癌Hep3B细胞株增殖,并可能通过上调[Ca2+]i诱导其凋亡。关键词:齐墩果酸;肝癌细胞Hep3B;细胞周期;细胞凋亡;细胞内钙离子浓度中图分类号R282.75文献
3、标志码AEffectsofOleanolicAcidonProliferationandApoptosisofHepatomaCellLineHep3BHuangKai-shun1,ZhuLian-lian2,LiuDan2,ZouLi2,GePu3,leiQi2,ChenZhi-qiong1,2*(1.InstituteofLifeSciences;2.DepartmentofPharmacyAnalysis,SchoolofPharmacy;3.DepartmentofPathophysiology,SchoolofBasicMedicine,Chon
4、gqingMedicalUniversity,Chongqing400016)Abstract:ObjectiveToinvestigatetheinfluenceofoleanolicacidontheproliferationofhumanhepatomaHep3BcellandtherelationshipbetweenapoptosisofHep3BcellandintracellularCa2+concentration([Ca2+]i).MethodsHep3Bcellsweretreatedwitholeanolicacidofdiffere
5、ntconcentrationsandstainedwithDAPI,morphologychangeswereobservedbyfluorescencemicroscopy;growthinhibitionratioesweremeasuredbytheMTTmethod;Cellcycle,percentagesofapoposisandintracellularCa2+concentrations([Ca2+]i)werecheckedbyflowcytometry;Linearcorrelationwasanalizedbetween[Ca2+]
6、iandthepercentageofapoposisResultsTheapoptosisofHep3Bcelloccurred;OleanolicacidofdifferentconcentrationsinhibitedtheproliferationofHep3Bcellsinadosedependentmannerfrom5μg/mLto100μg/mL,withIC50being86.04μg/mLand93.29μg/mLfor24hand48h,respectively.CellcyclewasterminatedatG2/Mphaseby
7、oleanolicacidandintracellularcalciumionconcentrationwassignificantlyincreased,atthemeantime,percentagesofapoposisandincreasesof[Ca2+]idependedontheconcentrationofoleanolicacid.[Ca2+]i;andpercentagesofapoptosishavelinearcorrelationinadosemannerfrom80μg/mLto150μg/mL.ConclusionOleano
8、licacidmayinhibitthegrowthofHep3B