中国实验动物学报
中國實驗動物學報
중국실험동물학보
Acta Laboratorium Animalis Scientia Sinica
2015年
5期
466-473
,共8页
三阴性乳腺癌%肿瘤抗药性%顺铂%动物模型%小鼠
三陰性乳腺癌%腫瘤抗藥性%順鉑%動物模型%小鼠
삼음성유선암%종류항약성%순박%동물모형%소서
Triple negative breast cancer%Neoplasm drug resistance%Cisplatin%Animal models%Mice
目的 建立一种耐药性稳定的三阴性乳腺癌4 T1耐药小鼠模型,为研究体内肿瘤耐药机制和逆转药物的筛选奠定实验基础. 方法 采用顺铂( DDP)低剂量诱导及体内外交叉致瘤结合的方法建立三阴性乳腺癌耐药小鼠模型;MTT法检测细胞耐药特性;实时荧光定量PCR法分析耐药相关基因MDR1、BCRP、MMP7及 GST-π表达差异;免疫组化分析耐药相关蛋白P-gp、BCRP、MMP7表达差异;蛋白印迹法检测磷酸化Akt(phosphorate-Akt, p-Akt)和总Akt( total-Akt,t-Akt)蛋白表达;小动物成像检测观察肿瘤生长情况. 结果 MTT显示建立的三阴性乳腺癌耐药4T1小鼠模型的耐药指数为12.84;耐药小鼠肿瘤组织中MDR1、BCRP、MMP7、GST-π基因mRNA 的表达量及P-gp、BCRP、MMP7蛋白的表达量均高于非耐药小鼠(P<0.01);Western blot显示,耐药小鼠肿瘤组织的p-Akt蛋白表达明显高于非耐药小鼠,t-Akt蛋白表达没有差异. 非耐药小鼠与耐药小鼠肿瘤组织生长速度未见明显区别(P>0.05). 分别给予这两种模型小鼠相同剂量的DDP治疗后,耐药小鼠对DDP的敏感性明显低于非耐药小鼠(P<0.01). 结论 初步建立了三阴性乳腺癌耐药4T1小鼠模型,为三阴性乳腺癌临床个体化治疗及耐药逆转研究等提供了良好的实验动物平台.
目的 建立一種耐藥性穩定的三陰性乳腺癌4 T1耐藥小鼠模型,為研究體內腫瘤耐藥機製和逆轉藥物的篩選奠定實驗基礎. 方法 採用順鉑( DDP)低劑量誘導及體內外交扠緻瘤結閤的方法建立三陰性乳腺癌耐藥小鼠模型;MTT法檢測細胞耐藥特性;實時熒光定量PCR法分析耐藥相關基因MDR1、BCRP、MMP7及 GST-π錶達差異;免疫組化分析耐藥相關蛋白P-gp、BCRP、MMP7錶達差異;蛋白印跡法檢測燐痠化Akt(phosphorate-Akt, p-Akt)和總Akt( total-Akt,t-Akt)蛋白錶達;小動物成像檢測觀察腫瘤生長情況. 結果 MTT顯示建立的三陰性乳腺癌耐藥4T1小鼠模型的耐藥指數為12.84;耐藥小鼠腫瘤組織中MDR1、BCRP、MMP7、GST-π基因mRNA 的錶達量及P-gp、BCRP、MMP7蛋白的錶達量均高于非耐藥小鼠(P<0.01);Western blot顯示,耐藥小鼠腫瘤組織的p-Akt蛋白錶達明顯高于非耐藥小鼠,t-Akt蛋白錶達沒有差異. 非耐藥小鼠與耐藥小鼠腫瘤組織生長速度未見明顯區彆(P>0.05). 分彆給予這兩種模型小鼠相同劑量的DDP治療後,耐藥小鼠對DDP的敏感性明顯低于非耐藥小鼠(P<0.01). 結論 初步建立瞭三陰性乳腺癌耐藥4T1小鼠模型,為三陰性乳腺癌臨床箇體化治療及耐藥逆轉研究等提供瞭良好的實驗動物平檯.
목적 건립일충내약성은정적삼음성유선암4 T1내약소서모형,위연구체내종류내약궤제화역전약물적사선전정실험기출. 방법 채용순박( DDP)저제량유도급체내외교차치류결합적방법건립삼음성유선암내약소서모형;MTT법검측세포내약특성;실시형광정량PCR법분석내약상관기인MDR1、BCRP、MMP7급 GST-π표체차이;면역조화분석내약상관단백P-gp、BCRP、MMP7표체차이;단백인적법검측린산화Akt(phosphorate-Akt, p-Akt)화총Akt( total-Akt,t-Akt)단백표체;소동물성상검측관찰종류생장정황. 결과 MTT현시건립적삼음성유선암내약4T1소서모형적내약지수위12.84;내약소서종류조직중MDR1、BCRP、MMP7、GST-π기인mRNA 적표체량급P-gp、BCRP、MMP7단백적표체량균고우비내약소서(P<0.01);Western blot현시,내약소서종류조직적p-Akt단백표체명현고우비내약소서,t-Akt단백표체몰유차이. 비내약소서여내약소서종류조직생장속도미견명현구별(P>0.05). 분별급여저량충모형소서상동제량적DDP치료후,내약소서대DDP적민감성명현저우비내약소서(P<0.01). 결론 초보건립료삼음성유선암내약4T1소서모형,위삼음성유선암림상개체화치료급내약역전연구등제공료량호적실험동물평태.
Objective To establish a cisplatin-resistant 4T1 mouse model of triple negative breast cancer .Meth-ods A drug resistant mice model was established with cisplatin ( DDP ) induction and in-vivo/in-vitro tumorigenic ap-proach.Its resistance characteristics were identified by MTT assay .Changes of drug resistance gene ( MDR1, BCRP, MMP7, GST-π) and protein ( P-gp, BCRP, MMP7) expression, and phosphorate-Akt and total-Akt protein expression were evaluated by real-time PCR, immunohistochemistry and western blot method , respectively.Small animal live imaging technology was applied to detect tumor growth .Results Resistance fold (RF) of cisplatin-resistant 4T1 mouse model was 12.84.The expression of MDR1, BCRP, MMP 7, GST-πmRNA and P-gp, BCRP, MMP 7 proteins in the resistant mice were higher than that in the non-resistant mice .The result of western blot showed that a statistically higher expression of p-Akt in resistant mice than that in non-resistant mice at protein levels (P<0.01).No significant difference of tumor growth rate was observed between non-resistant and resistant mice ( P>0.05 ) .Given same dose of DDP , resistant mice showed lower sensitivity than non-resistant mice significantly (P<0.01).Conclusions We have successfully established a cis-platin-resistant triple negative breast cancer model in mice , which provides a new platform for further study on chemoresis-tant reversal strategy and individualized clinical treatment of this disease .
