结肠癌皮下移植瘤模型方法的优化  

The optimization of colon cancer xenograft model establishment

作　　者：肖开提.阿不都哈德尔 王理[1] 张文斌[1] 王云海[1] 

机构地区：[1]新疆医科大学第一附属医院胃肠外科,乌鲁木齐830054

出　　处：《新疆医科大学学报》2015年第5期586-589,共4页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区自然科学基金(2014211C036)

摘　　要：目的通过分组筛选建立结肠癌皮下移植瘤模型的最优条件,并以此为瘤源建立结肠癌皮下移植瘤模型。方法根据细胞介质和注射部位的不同,将人结肠癌细胞株HT29分别用PBS和无血清培养基重悬后注射在BALB/c小鼠腋下和颈部皮下,建立第1代皮下移植瘤模型。以此为移植瘤源,利用插块法、酶解法及匀浆法,建立第2代皮下移植瘤模型,选择筛选后最优选的方法建立第3代皮下移植瘤模型,观察肿瘤生长及转移等情况,采用组织病理学、免疫组织化学、流式细胞技术对移植瘤进行检测。结果成功筛选建立了具有可操作性、高重复性的人结肠癌皮下移植瘤模型。移植瘤组织切片观察显示肿瘤为腺癌I～Ⅱ级,免疫组化显示CK19和P53呈阳性至强阳性表达,流式细胞术检查分析DNA倍体为异倍体。结论本实验利用人结肠癌细胞株HT29成功筛选并建立了合适的结肠癌皮下移植瘤模型,该模型为结肠癌转移的下一步生物学研究提供了一个良好的技术平台。Objective To establish a colon cancer xenograft model through the packet filtering optimum conditions.Methods According to different cell media and the injection sites,the HT29 human colon cancer cell lines were washed with PBS and resuspended in serum-free medium after injection in BALB/C mice armpit and neck skin,first-generation xenograft model was established.That established model was taken as an xenograft source,the interpolation block method and homogenized enzymatic method were used to build a second-generation xenograft model.The most preferred method of screening was selected to establish the third generation of subcutaneous xenograft model.Tumor growth conditions and metastasis were observed; histopathology,immunohistochemistry and flow cytometry were adopted to detect the transplanted tumor.Results The successful establishment of colon cancer xenograft model was of operability and high repeatability.Transplanted tumor tissue biopsies showed that the tumor was adenocarcinoma of grade Ⅰ-Ⅱ.Immunohistochemistry showed positive expression of CK19 and P53.The check of DNA ploidy by flow cytometry showed aneuploid.Conclusion A proper screening and colon cancer xenograft modelw was successfully established by using human colon cancer cell line HT29 with BALB/c mice,which provided a good technical platform for the further biological research on colon cancer metastasis.

关 键 词：结肠癌 HT29 皮下移植瘤 动物模型 

分 类 号：R735[医药卫生—肿瘤]