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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 subcutaneous xenograft model of colon cancer by establishing a subgroup of colon cancer subcutaneous xenograft models. Methods Human colon cancer cell line HT29 was resuspended with PBS and serum-free medium respectively and then injected into the subcutaneous and neck of BALB / c mice subcutaneously to establish a primary subcutaneous xenograft model . The second generation of subcutaneous xenograft model was established by inserting block method, enzymolysis method and homogenate method. The third generation of subcutaneous xenograft model was established by selecting the best method after screening, and observed the growth and metastasis of tumor The situation, using histopathology, immunohistochemistry, flow cytometry on the detection of xenografts. Results Successful screening of subcutaneously transplanted human colon carcinoma models with operable and highly reproducible results. Tumor tissue sections were observed for grade I ~ II of adenocarcinoma. Immunohistochemistry showed positive and strongly positive expression of CK19 and P53. DNA was analyzed by flow cytometry analysis of aneuploidy. Conclusion This study successfully screened human colon cancer cell line HT29 and established a suitable model of subcutaneously transplanted colon cancer. This model provides a good technical platform for the further biological study of colon cancer metastasis.