中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2015年
24期
1910-1914
,共5页
刘艳%张霄蓓%刘鹏%张瑾
劉豔%張霄蓓%劉鵬%張瑾
류염%장소배%류붕%장근
乳腺肿瘤%干细胞%mTOR抑制剂%依维莫司
乳腺腫瘤%榦細胞%mTOR抑製劑%依維莫司
유선종류%간세포%mTOR억제제%의유막사
Breast neoplasms%Stem cell%mTOR inhibitor%Everolimus
目的 观察抑制PI3 K/Akt/mTOR信号通路中mTOR表达对曲妥珠单抗耐药的乳腺癌干细胞(BCSCs)的作用及对靶向治疗药物敏感性的影响.方法 使用流式仪分选ESA+ CD44+CD24-/low的人乳腺癌细胞系MCF-7,MDA-MB-231,BT474的BCSCs亚群,并对其进行克隆培养.通过MTT检测细胞增殖、软琼脂克隆实验细胞克隆形成能力.建立乳腺癌干细胞裸鼠移植瘤模型,记录小鼠体重及移植瘤大小以了解药物对BCSCs的体内作用影响;利用SP免疫组化方法检测不同药物作用下移植瘤中Ki67、CD31、AKT1、phospho-Akt(Thr308)的表达水平改变.结果 在体外,依维莫司对BCSCs存在一定抑制作用,抑制50％的原代BCSCs所需依维莫司浓度(5 170 nmol/L)是MCF-7中未分选细胞(NSC) (225 nmol/L)的23倍.相较于各自NSC的IC50,MCF-7、MDA-MB-231、BT474干细胞对依维莫司(2 369、9 773、2 054 nmol/L)的IC50分别是其NSC的IC50的42、38和29倍.可诱导BCSCsG0～ G1期的周期阻滞,发生早期凋亡,减少BCSCs体外克隆形成能力.在体内,依维莫司可有效抑制BCSCs移植瘤增长,Ki67、CD31蛋白表达显著降低(10.1％、16.3％),AKT1,phospho-Akt(Thr308)表达上调.结论 应用mTOR抑制剂依维莫司可有效抑制BCSCs的生长增殖.
目的 觀察抑製PI3 K/Akt/mTOR信號通路中mTOR錶達對麯妥珠單抗耐藥的乳腺癌榦細胞(BCSCs)的作用及對靶嚮治療藥物敏感性的影響.方法 使用流式儀分選ESA+ CD44+CD24-/low的人乳腺癌細胞繫MCF-7,MDA-MB-231,BT474的BCSCs亞群,併對其進行剋隆培養.通過MTT檢測細胞增殖、軟瓊脂剋隆實驗細胞剋隆形成能力.建立乳腺癌榦細胞裸鼠移植瘤模型,記錄小鼠體重及移植瘤大小以瞭解藥物對BCSCs的體內作用影響;利用SP免疫組化方法檢測不同藥物作用下移植瘤中Ki67、CD31、AKT1、phospho-Akt(Thr308)的錶達水平改變.結果 在體外,依維莫司對BCSCs存在一定抑製作用,抑製50％的原代BCSCs所需依維莫司濃度(5 170 nmol/L)是MCF-7中未分選細胞(NSC) (225 nmol/L)的23倍.相較于各自NSC的IC50,MCF-7、MDA-MB-231、BT474榦細胞對依維莫司(2 369、9 773、2 054 nmol/L)的IC50分彆是其NSC的IC50的42、38和29倍.可誘導BCSCsG0～ G1期的週期阻滯,髮生早期凋亡,減少BCSCs體外剋隆形成能力.在體內,依維莫司可有效抑製BCSCs移植瘤增長,Ki67、CD31蛋白錶達顯著降低(10.1％、16.3％),AKT1,phospho-Akt(Thr308)錶達上調.結論 應用mTOR抑製劑依維莫司可有效抑製BCSCs的生長增殖.
목적 관찰억제PI3 K/Akt/mTOR신호통로중mTOR표체대곡타주단항내약적유선암간세포(BCSCs)적작용급대파향치료약물민감성적영향.방법 사용류식의분선ESA+ CD44+CD24-/low적인유선암세포계MCF-7,MDA-MB-231,BT474적BCSCs아군,병대기진행극륭배양.통과MTT검측세포증식、연경지극륭실험세포극륭형성능력.건립유선암간세포라서이식류모형,기록소서체중급이식류대소이료해약물대BCSCs적체내작용영향;이용SP면역조화방법검측불동약물작용하이식류중Ki67、CD31、AKT1、phospho-Akt(Thr308)적표체수평개변.결과 재체외,의유막사대BCSCs존재일정억제작용,억제50％적원대BCSCs소수의유막사농도(5 170 nmol/L)시MCF-7중미분선세포(NSC) (225 nmol/L)적23배.상교우각자NSC적IC50,MCF-7、MDA-MB-231、BT474간세포대의유막사(2 369、9 773、2 054 nmol/L)적IC50분별시기NSC적IC50적42、38화29배.가유도BCSCsG0～ G1기적주기조체,발생조기조망,감소BCSCs체외극륭형성능력.재체내,의유막사가유효억제BCSCs이식류증장,Ki67、CD31단백표체현저강저(10.1％、16.3％),AKT1,phospho-Akt(Thr308)표체상조.결론 응용mTOR억제제의유막사가유효억제BCSCs적생장증식.
Objective To test the antitumor effects of the mTOR inhibitor everolimus in breast cancer stem cells and total cells in vitro and in vivo.Methods In vitro studies,we sorted ESA + CD44 + CD24-/low cells as stem cells using flow cytometry from primary breast cancer cells.Apoptosis and the cell cycle distributions of stem cells were examined by flow cytometry.The tumorigenicity of stem cells after treatment was investigated by soft agar colony formation assays.BALB/c mice were injected with stem calls and the different treatments were administered.After necropsy,the expression of Ki67,CD31,AKT1,and phospho-AKT (Thr308) was analyzed by immunohistochemistry.Results Treatment with everolimus resulted in growth inhibition of all stem cells in a dose-dependent manner.An increase in G0-G1 cell cycle arrest and an increased popalation of cells in early apoptosis were seen in everolimus treatment.In vivo,the volumes of the xenograft tumors significantly decreased in everolimus alone group compared to control group.Conclusions Everolimus could inhibit the growth of both total breast cancer cells and sorted stem cells in vitro and in vivo.
