卵巢癌细胞对顺铂耐药及其逆转的实验研究

ExperimentalStudyontheMechanismofCisplatinResistanceandItsReversioninHumanOvarianCancer

目的：探明卵巢癌对顺铂耐药的发生机理，并筛选出逆转其耐药性的有效调节剂。方法：建立对顺铂耐药的人卵巢癌细胞系ＳＫＯＶ３／ｃｐ，采用微囊包裹肿瘤细胞技术建立人类卵巢癌小鼠异种移植耐药模型。比较耐药细胞和非耐药细胞生化特征的差异，采用相应的耐药调节剂来进行耐药逆转实验。结果：在ＳＫＯＶ３细胞内，铂（Ｐｔ）累积浓度、Ｐｔ－ＤＮＡ加合物、ＤＮＡ链间交联指数（ＩＳＣ）分别是ＳＫＯＶ３／ｃｐ细胞的５．１、２．４和４．８倍（Ｐ＜０．０１，Ｐ＜０．０５，Ｐ＜０．０１）；二性霉素Ｂ（ＡｍＢ）和新生霉素（ＮＶＢ）能提高ＳＫＯＶ３／ｃｐ细胞内铂浓度或Ｐｔ－ＤＮＡ加合物浓度，从而完全或部分逆转ＳＫＯＶ３／ｃｐ对顺铂的耐药性。结论：导致ＳＫＯＶ３／ｃｐ对顺铂耐药的主要原因是细胞内药物浓度下降和对Ｐｔ－ＤＮＡ清除能力增强。ＡｍＢ和ＮＶＢ能够逆转ＳＫＯＶ３／ｃｐ对顺铂的耐药性。

Objective:Toexplorethemechanismofcisplatinresistanceanditsreversioninhumanovariancancer.Methods:Axenograftedcisplatinresistantmicemodelofhumanovariancancer,SKOV3/cp,wasdevelopedbythemicroencapsulatedtechnique.Themultiplechangesofbio-chemicalmarkersinthemodelweredetermined,andvariousmodulatorsforreversionweretested.Results:IntracelularplatinumacumulationinSKOV3was5.1times,Pt-DNAadducts2.4timesandinterstrandcroslinkofDNA(ISC)4.8timesofthoseincisplatin-resistantcelline,SKOV3/cp.ThesechangesinSKOV3/cpcouldnotbereversedbyverapamil.AmphotercinB(AmB)andNovobiocin(NVB)couldraisetheconcentrationsofplatinumandPt-DNAadductsinSKOV3/cp,resultingincompleteorpartialreversionofcisplatin-resistanceofSKOV3/cp.Therewerenodiferencesintotalglutathione(GSH)levelandinsensitivitytoCdCl2betweenSKOV3andSKOV3/cp.Conclusions:ItissuggestedthattheprimaryfactorcausingSKOV3/cpresistancetocisplatinisthereductionofintracelularplatinumaccumulationandtheaugmentationoftheabilitytoremovePt-DNAadducts.Theresistanceisnotconsideredtobeassociatedwiththemultidrugresistant,GSH,metalothioneinsystems.AmBandNVBcanovercomecisplatinresistanceofSKOV3/cpinvitroandinvivo.