结肠癌皮下移植瘤模型方法的优化.docVIP

  结肠癌皮下移植瘤模型方法的优化# 张文斌，王理，依里牙尔·依里哈木，高华* （新疆医科大学第一附属医院胃肠外科 新疆乌鲁木齐 830000） 5 10 15 20 25 30 35 40 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，通过观察肿瘤生长及转移等情况，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～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。 关键词：结肠癌;HT29;皮下移植瘤;动物模型 中图分类号：R735.3+5 Optimization of colon cancer xenograft model establishment method Zhang Wenbin, Wang li, Eliyar`Elham, Gao Hua (The First Clinical Medical School of Xinjiang Medical University,Urumqi, 830000,China) Abstract: Objective: To establish a colon cancer xenograft model through the packet filtering optimum conditions. Methods: According to different cell media and the injection site, 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, build first-generation xenograft model, as a xenograft source, using interpolation block method, and homogenized enzymatic method, a second-generation xenograft model, choose the most preferred method of screening after the establishment of the third generation of subcutaneous xenograft model, by observing tumor growth conditions and metastasis, histopathology, immunohistochemistry, flow cytometry to detect transplanted tumor. Results: The success of established operability, high repeatability colon cancer xenograft model. Observed tumor biopsy showed that the tumor was adenocarcinoma I ~ Ⅱ grade. Immunohistochemistry showed positive expression of CK19 and P53. Flow cytometry analysis of DNA ploidy check aneuploid.Conclusion: In this study, we successfully established a proper screening and colon cancer xenograft model use human colon cancer cell line HT29with BALB/C mice,and provid