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《P-糖蛋白1对宫颈癌化疗耐药的影响及机制-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在应用文档-天天文库。
1、中南医学科学杂志2014年9月第42卷第5期447文章编号:2095—1116(2014)05—0447—05·基础医学·P一糖蛋白1对宫颈癌化疗耐药的影响及机制童文娟。孙少卫(1.南华大学附属第一医院妇产科,湖南衡阳421001;2.南华大学生命科学研究中心)摘要:目的探讨P一糖蛋白l(P.gP)在宫颈癌细胞对依托泊苷耐药中的作用机制。方法观察不同分期宫颈癌组织和慢性宫颈炎组织中P—gP的表达差异。取Hela细胞分别用azithromycin和cyclosporineA处理6h后,再与依托泊苷孵育不同时间,高效液相色谱
2、(HPLC)检测细胞内依托泊苷含量。将Hela细胞培养于培养皿或transwell小室中,用依托泊苷处理30min,再分别与azithromycin和cyelosporine共孵育,检测transwell下室培养液中依托泊苷含量,同时检测各组细胞凋亡情况及凋亡相关因子Bcl一2和Bax的表达水平。结果P-gP在宫颈癌组织中表达显著高于宫颈炎组织。azithromycin可增强依托泊苷对Hela细胞的损伤作用,并呈浓度依赖性地上调Bax表达,降低Bc1.2水平,而cyclosporineA则对Hela细胞有保护作用。Tra
3、nswell培养实验发现,azithromycin增加Hela细胞层对依托泊苷的通透性,而cyclosporineA则降低其通透性。结论P—gP促使Hela细胞泵出依托泊苷,减轻细胞损伤,因此介导了宫颈癌对依托泊苷的耐药性。关键词:P.糖蛋白1;宫颈癌;依托泊苷;Hela细胞中图分类号:R737.33文献标识码:AP-glycoprotein1MdiatedtheResistanceofCervicalCancertoEtoposideTONGWenjuan,SUNShaowei(DepartmentofGynecolo
4、gyandObstetrics,theFirstAffiliatedHospital,Unive~ityofSouthChina,Hengyang,Hunan421001,China)Abstract:ObjectiveToinvestigatetherolesofP-glycoprotein1onetoposideresistanceofcervicalcancercells.MethodTheexpressionofP-glycoprotein1incervicalcancerandchroniccervicitis
5、tissuewereobservedatdiferentperi-·ods.Thehelacellsweretreatedwithazithromycin(P—glyeoprotein1activator)orcyclosporineA(P—glycoprotein1inhibi—tor)for6hours,thenincubatedwithetoposidefordifferenttimes.ThecontentofetoposideinhelacellswasdetectedbyHPLC.Helacellswereg
6、rownonaTranswellinserttoobtainatightmonolayerandpreincubatedwithetoposidefor30mi-nutes,thenincubatedforanother12hafteradditionofazithromyein,orcyelosporineintheapicalreservoir,HPLCwasusedtodetectthecontentofetoposideinbottomreservoir.Helacellsweretreatedwithazith
7、romycinorcyclosporinefor24hafterpreincubatedwithetoposide,thecellviabilitywasdeterminedbyMTFassay,Westernblotwasusedtoanalyzetheexpres-sionofapoptosisrelatedproteinsBcl-2andBax.ResultsTheexpressionofP—glycoprotein1wassignificantlyhigherincervicalcancertissuesthan
8、incervieitis,anditsexpressionlevelwascloselyrelatedwithcancerstaging.Azithromycinen—hancedthecytotoxiceffectofetoposideonhelacells,increasedtheexpressionofBax,
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