增免抑瘤方及有效组分对卵巢癌干细胞裸鼠移植瘤Notch/Jagged1信号通路的影响  

Effect of Zengmian Yiliu Formula and Its Effective Components on Notch/Jagged1 Signaling Pathway of Ovarian Cancer Stem Cell Transplanted Tumor in Nude Mice

作　　者：张婕 梁程程 杨爱仲 齐聪[3] ZHANG Jie;LIANG Cheng-cheng;YANG Ai-zhong;QI Cong(Longhua Hospital,Shanghai University of Traditional Chinese Medicine(TCM),Shanghai 200032,China;Graduate School,Shanghai University of TCM,Shanghai 200120,China;Shuguang Hospital,Shanghai University of TCM,Shanghai 200120,China)

机构地区：[1]上海中医药大学附属龙华医院,上海200032 [2]上海中医药大学研究生院,上海200120 [3]上海中医药大学附属曙光医院,上海200120

出　　处：《中国实验方剂学杂志》2020年第15期46-53,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金青年科学基金项目(81603645);上海市卫生和计划生育委员会科研课题青年项目(20164Y0243)。

摘　　要：目的:观察增免抑瘤方(ZMYL)及其有效组分(PAWU)对卵巢癌干细胞裸鼠移植瘤的生长抑制作用及对肿瘤组织Notch信号受体(Notch)/Notch信号配体1(Jagged1)信号通路的影响。方法:无血清悬浮培养卵巢癌干细胞,建立卵巢癌干细胞裸鼠移植瘤模型,分为模型组、增免抑瘤方组(ZMYL,36 g·kg^-1),有效组分组(PAWU,5.8 g·kg^-1),顺铂组(DDP,2.5 mg·kg^-1),有效组分联合顺铂组(PAWU,5.8 g·kg^-1+DDP,2.5 mg·kg^-1)。成功造模后,灌胃给药21 d。观察增免抑瘤方及有效组分对裸鼠瘤重的影响;苏木素-伊红(HE)染色观察肿瘤细胞的形态变化;免疫组化、实时荧光定量聚合酶链式反应(Real-time PCR)检测肿瘤组织中Notch信号受体1(Notch1),Notch信号配体1(Jagged1),发状分裂相关增强子1(Hes1)蛋白和mRNA的表达水平。结果:各给药组对裸鼠皮下移植瘤的生长均具有抑制作用,ZMYL组,PAWU组,DDP组,PAWU+DDP组抑瘤率分别为35.91%,32.94%,57.65%,69.05%,与模型组比较,ZMYL组,PAWU组,DDP组,PAWU+DDP组瘤重明显降低(P<0.05),与PAWU组比较,PAWU+DDP组瘤重明显降低(P<0.05);免疫组化结果表明,与模型组比较,ZMYL组,PAWU组,DDP组,PAWU+DDP组Notch1,Jagged1蛋白阳性表达明显减少(P<0.05),ZMYL组,DDP组,PAWU+DDP组Hes1蛋白阳性表达明显减少(P<0.05);与PAWU+DDP组比较,ZMYL组,PAWU组,DDP组Notch1,Jagged1,Hes1蛋白阳性表达明显上调(P<0.05);Real-time PCR结果表明,与模型组比较,ZMYL组,PAWU组,DDP组,PAWU+DDP组Notch1 mRNA表达率明显减少(P<0.05),ZMYL组,PAWU组,PAWU+DDP组的Jagged1,Hes1 mRNA表达率明显减少(P<0.05),与DDP组比较,PAWU+DDP组Notch1,Jagged1,Hes1 mRNA表达率明显减少(P<0.05)。结论:增免抑瘤方及有效组分可以抑制卵巢癌干细胞裸鼠移植瘤的生长,有效组分与顺铂联合应用有明显的协同增效作用,其机制可能与抑制Notch/Jagged1信号通路的激活相关。Objective:To observe the effect of Zengmian Yiliu(ZMYL)formula and its effective components(PAWU)on the growth inhibition of ovarian cancer stem cell transplanted tumor in nude mice and the Notch signal receptor(Notch)/Notch signal ligand 1(Jagged1)signal pathway in tumor tissue.Method:Ovarian cancer stem cells were cultured in serum-free suspension to establish the transplanted tumor model of ovarian cancer stem cells in nude mice,and then divided into model group,ZMYL group(36 g·kg^-1),PAWU group(5.8 g·kg^-1),cisplatin(DDP)group(2.5 g·kg^-1),and PAWU(5.8 g·kg^-1)+DDP group(2.5 mg·kg^-1).After successful modeling,the drugs were given by gavage for 21 days.To observe the effect of Zengmian Yiliu decoction and its effective components on tumor weight in nude mice,the morphological changes of tumor cells were observed under light microscope,immunohistochemistry and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)were used to detect the expressions of Notch 1,Jagged1,Hairy and enhancer of split 1(Hes1)protein and mRNA in tumor tissues.Result:The tumor inhibition rates of ZMYL,PAWU,DDP and combination groups were 35.91%,32.94%,57.65%and 69.05%,respectively.Compared with the model group,the tumor weight of ZMYL group,PAWU group,DDP group and combination groups decreased significantly(P<0.05).Compared with PAWU group,the tumor weight of combination groups decreased significantly(P<0.05).Immunohistochemistry showed that compared with the model group,the positive expressions of Notch1,Jagged1 in ZMYL group,PAWU group,DDP group and combination groups were down-regulated(P<0.05),and the positive expressions of Hes1 in ZMYL group,DDP group and combination groups were down-regulated(P<0.05).Compared with combination groups,the positive expressions of Notch1,Jagged1 and Hes1 in ZMYL group,PAWU group,DDP group were up-regulated(P<0.05).Real time PCR showed that compared with the model group,the expressions of Notch1 mRNA in ZMYL group,PAWU group,DDP group and combination groups decreased signif

关 键 词：增免抑瘤方 有效组分 裸鼠 卵巢癌干细胞 Notch信号受体(Notch)/Notch信号配体1(Jagged1) 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]