基于过表达羧酸酯酶的人骨髓间充质干细胞联合LY2334737对膀胱癌的抑制作用研究  

Inhibitory Effect of Human Bone Mesenchymal Stromal Cells Overexpressing Carboxylesterases Combined with LY2334737 on Bladder Cancer

作　　者：李杨东 范毛川[1] 李卫胜 窦启锋[1] Li Yangdong;Fan Maochuan;Li Weisheng;Dou Qifeng(Department of Urology,the First Afiliated Hospital of Xinxiang Medical College,Xinxiang 453199,Henan,China;Department of Nursing,Shizhen College,Guizhou University of Traditional Chinese Medicine,Guiyang 550200,Guizhou,China;Department of Urology,the First Afiliated Hospital of Henan University of Chinese Medicine,Xinxiang 571199,Henan,China)

机构地区：[1]新乡医学院第一附属医院泌尿外科,河南新乡453199 [2]贵州中医药大学时珍学院护理学部,贵阳550200 [3]河南中医药大学第一附属医院泌尿外科,郑州571199

出　　处：《肿瘤预防与治疗》2024年第10期831-842,共12页Journal of Cancer Control And Treatment

基　　金：河南省科技攻关项目(编号:232102310349);新乡市灾后重建科技专项项目(编号:21CJ002)。

摘　　要：目的:探讨过表达羧酸酯酶(carboxylesterases,CES)的人骨髓间充质干细胞(human bone mesenchymal stromal cells,hBMSCs)联合LY2334737对裸鼠膀胱癌治疗的影响。方法:将腺病毒介导过表达的人CES2转染hBMSCs,分为空白组(不作任何处理)、空载体组(转染空载体)和转染组(转染过表达CES2腺病毒),流式细胞仪检测转染效率,荧光显微镜观察CES2的表达,Western blot及RT-PCR方法检测转染hBMSCs的效果。hBMSCs转染48 h后,用LY2334737(1μmol/L)处理72 h。将处理的细胞分为空白组(hBMSCs)、对照组(Ad-hBMSCs+LY2334737)、试验组(CES2-hBMSCs+LY2334737),采用CCK-8法及流式细胞仪检测hBMSCs的细胞增殖、凋亡情况。采用Transwell小室将人膀胱癌细胞TCCSUP与hBMSCs共培养,将其分为5组:空白组、空载体组、转染组、Ad-hBMSCs+LY2334737组、CES2-hBMSCs+LY2334737组,CCK-8法及流式细胞仪检测TCCSUP细胞的增殖、凋亡情况。将60只雌性裸鼠随机分为6组:空白组(仅注射TCCSUP细胞)、对照组(注射TCCSUP细胞和Ad-hBMSCs)、试验组(注射TCCSUP细胞和CES2-hBMSCs)、LY2334737组(注射TCCSUP细胞+口服LY2334737)、Ad-hBMSCs+LY2334737组(注射TCCSUP细胞和Ad-hBMSCs+口服LY2334737)、CES2-hBMSCs+LY2334737组(注射TCCSUP细胞和CES2-hBMSCs+口服LY2334737),裸鼠麻醉后,于背中线皮下注射TCCSUP细胞。分别在注射TCCSUP细胞1周及2周后,在肿瘤周围注射Ad-hBMSCs或CES2-hBMSCs。在第一次注射BMSCs 1 h后,LY2334737口服给药14天,每天1次。LY2334737末次治疗结束1天后处死动物称瘤重量,测量瘤体积,对肿瘤组织进行HE染色。结果:流式细胞术检测过表达CES2腺病毒转染组的转染率为99.56%;RT-PCR结果表明,与空白组和空载体组相比,转染组CES2的mRNA表达量增加,差异具有统计学意义(t=27.642,P<0.0001;t=30.300,P<0.0001);Western blot结果发现,与空白组和空载体组相比,过表达CES2转染组的CES2蛋白量表达升高(t=9.678,P=0.0006;t=9.516,P=0.0007),说明腺病毒过表�Objective:To investigate the effect of human bone mesenchymal stromal cells(hBMSCs)overexpressing carboxylesterases(CES)combined with LY2334737 on the treatment of bladder cancer in nude mice.Method:Adenovirus-mediated overexpressed human CES2 transfected hBMSCs were assigned to the blank group(without any treatment),the empty carrier group(transfected with empty carrier)and the transfection group(transfected with CES2 overexpressed adenovirus).The transfection efficiency was detected by flow cytometry,and the expression of CES2 was observed by fluorescence microscope.Western blot and reverse transcription polymerase chain reaction(RT-PCR)were used to detect the transfection effect of hBMSCs.After transfection for 48 h,hBMSCs were treated with LY2334737(1μmol/L)for 72 h.The treated cells were assigned to the blank group(hBMSCs),the control group(Ad-hBMSCs+LY2334737)and the experimental group(CES2-hBMSCs+LY2334737).The proliferation and apoptosis of hBMSCs were detected by CCK-8 method and flow cytometry.Human bladder cancer cells TCCSUP and hBMSCs were co-cultured using Transwell chamber and divided into 5 groups:The blank group,the empty carrier group,the transfection group,the Ad-hBMSCs+LY2334737 group and the CES2-hBMSCs+LY2334737 group.The proliferation and apoptosis of TCCSUP cells were determined by CCK-8 method and flow cytometry.Sixty female nude mice were randomly divided into 6 groups:The blank group(injected with TCCSUP cells only),the control group(TCCSUP cells and Ad-hBMSCs injected),the experimental group(TCCSUP cells and CES2-hBMSCs injected),the LY2334737 group(TCCSUP cells injected+oral LY2334737),the Ad-hBMSCs+LY2334737 group(injected TCCSUP cells and Ad-hBMSCs+oral LY2334737),the CES2-hBMSCs+LY2334737 group(injected TCCSUP cells and CES2-hBMSCs+oral LY2334737).After nude mice were anaesthetized,TCCSUP cells were injected subcutaneously into the dorsal midline.Ad-hBMSCs or CES2-hBMSCs were injected around the tumor 1 week and 2 weeks after the injection of TCCSUP cells.1 h after BMSCs was initi

关 键 词：膀胱癌 人骨髓间充质干细胞 羧酸酯酶2 LY2334737 凋亡 增殖 

分 类 号：R735.7[医药卫生—肿瘤]