摘要
目的探讨利用裸鼠乳腺癌荷瘤模型转染解聚素-金属蛋白酶17(ADAM17)-shRNA的骨髓间充质干细胞(BMSCs)对乳腺癌移植瘤的抑制效果。方法将人乳腺癌细胞(MCF-7)接种于30支裸鼠皮下,14 d后乳腺癌荷瘤裸鼠模型成功构建。随机分为:转染组(注射1×10~6/ml转染ADAM17-shRNA的BMSCs)、BMSCs组(注射1×10~6/ml BMSCs)和对照组(注射空白PBS)。采用贴壁筛选法分离、培养3周龄雄性SD大鼠的BMSCs,利用慢病毒介导将ADAM17-shRNA转染第3代培养的BMSCs,将其通过尾静脉注射给各组荷瘤裸小鼠(0.1 ml/次)3 d重复一次,共5次,观察各组裸小鼠肿瘤的生长状况。抑瘤实验16 d后处死裸鼠,HE染色法观察各组瘤体的形态结构,免疫组化法检测各组瘤体ADAM17、表皮细胞生长因子受体(EGFR)和细胞增殖核抗原(Ki-67)的表达。结果转染组、BMSCs组、对照组裸鼠瘤体体积分别为(375.09±20.72)、(768.85±35.55)、(783.75±27.30)mm3,转染组相较于对照组移植瘤体积缩小,差异有统计学意义(P<0.05),而BMSCs组差异无统计学意义。HE染色镜下可见,与对照组相比转染组可见瘤体组织大片坏死区域、坏死区域细胞崩解、细胞结构消失,而BMSCs组与对照组差异无统计学意义。转染组ADAM17、EGFR、Ki-67蛋白与对照组相比表达均降低,差异有统计学意义(P<0.05);BMSCs组和对照组相比较,差异无统计学意义。结论 ADAM17-shRNA通过BMSCs介导可靶向归巢至裸鼠乳腺癌移植瘤并发挥抑制裸鼠乳腺癌ADAM17、EGFR和Ki-67表达的作用。
Objective To explore expressions of ADAM17,EGFR,and Ki-67 in implanted breast cancer on nude mouse treated by bone marrow stem cells（ BMSCs） transfected with a disintegrin and metalloprotease 17（ ADAM17）-shRNA. Methods Thirty nude mice were inoculated subcutaneously with MCF-7 cells to construct the nude mice model of breast cancer. They were randomly divided into 3 groups： transfection group（ 1 × 10^6/ml BMSCs transfected ADAM17-shRNA）,BMSCs group（ 1 × 106/ml BMSCs） and control group（ blank PBS）. The BMSCs of male SD rats aged 3 w were isolated and cultured by adherence screening method. The third generation of BMSCs were transfected with ADAM17-shRNA by lentivirus,which were injected into nude mice by tail vein（ 0. 1 ml for each time,repeated once every 3 day for 5 times）. All animals were sacrifieced on 16 th day after treatment.The morphology of tumor were observed by HE staining,and expressions ADAM17,epidermal growth factor receptor（ EGFR） and cell proliferating nuclear antigen Ki-67（ Ki-67） of tumor was detected by immunohistochemistry.Results The implanted tumor volumes of transfection group,BMSCs group and control group was（ 375. 09 ±20. 72）,（ 768. 85 ± 35. 55）,（ 783. 75 ± 27. 30） mm3,respectively. The implanted tumor volumes of transfection group was significant reduced compared with BMSCs group and control group（ P〈 0. 05）. HE staining showed that the large area of necrosis of tumor tissue was seen in the transfection group,and the necrotic area cells were disintegrated and the cell structure disappeared. But there was no significant change between BMSCs group and control group. Conpared with control group,expressions of ADAM17,EGFR and Ki-67 protein in transfection group were markedly decreased（ P 〈0. 05）. There was no statistical difference in expressions of ADAM17,EGFR and Ki-67 protein between BMSCs group and control group. Conclusion ADAM17-shRNA can target homing to breast cancer xenograft in nude mouse mediated by BMSCs and inhibit t
作者
贾文婷
陈国福
张雪鹏
张笑博
孙影
赵莹
Jia Wenting;Cheng Guofu;Zhang Xuepeng(Dept of Surgical Oncology,Affiliated Hospital of North China Universityof Science and Technolo;,TangsPan 06300)
出处
《安徽医科大学学报》
CAS
北大核心
2018年第4期589-595,共7页
Acta Universitatis Medicinalis Anhui
基金
河北省自然科学基金(编号:C2010001767)
唐山市科学技术研究与发展计划项目(编号:14130256B)
华北理工大学研究生创新项目(编号:2017S43)
