大肠癌HT-29细胞亚系与母系转移能力和相关因子的比较

Comparative analysis of metastatic variants from the colorectal tumor cell line HT-29

目的 :探讨大肠癌细胞转移能力与肿瘤相关因素的关系。方法 :用结肠癌 HT- 2 9亚系 HT- 2 9c和 HT- 2 9d细胞在裸大鼠体内建立转移模型 ,比较其与母系的转移能力。用 ELISA法测定 3个大肠癌细胞系的尿激酶型纤溶酶原激活因子 (u PA)及纤溶酶原抑制剂 - 1(PAI- 1)含量 ;用免疫组化技术检测癌胚抗原和 3-磷酸肌醇激酶(PI3- Kinase)在体内外的表达 ;用流式细胞仪测定癌胚抗原表达。结果 :在裸鼠体内 HT- 2 9d细胞的肝转移率明显高于母系 HT- 2 9细胞 ,转移累及的脏器增多。在体外培养中 ,HT- 2 9d细胞的 u PA及 PAI- 1含量明显高于 HT- 2 9和 Wi Dr细胞。在裸鼠体内 ,HT- 2 9d细胞的 PI3- Kinase表达明显高于 HT- 2 9和 Wi Dr细胞。结论 :经裸鼠体内筛选的大肠癌 HT- 2 9细胞亚系表现出增强的肿瘤转移能力。UPA、PAI- 1的含量及 PI3- Kinase的表达与肿瘤转移能力有关。

Objective: To analyze the relationship between metastatic potential and related facters of colorectal tumor cell lines. Methods: The variants HT-29c and HT-29d cell lines derived from the selection of HT-29 cells were injected into nude rats and the metastatic potential of the two tumor cell variants was analyzed. Expression of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor type 1 (PAI-1) were measured with ELISA in vitro in colorectal carcinoma cell lines WiDr, HT-29 and HT-29d. Expression of carcinoembryonic antigen (CEA) and phosphoinositide 3-kinase (PI3-Kinase) were determined with immunohistochemistry, (IHC) in vitro and in vivo in WiDr, HT-29 and HT-29d cell lines. In addition, CEA expression was demonstrated with fluorescence activated cell sorter (FACS) in vitro. Results: The liver metastasis rate of the variant HT-29d (with 4 cycles of selection), increased as compared with that of parental HT-29 cells and that of variant HT-29b cells (with 2 cycle of selection). The uPA concentration of variant HT-29d cell line was significantly higher than that of the non-metastatic WiDr and the low metastatic HT-29 cells ( P <0.05). The variant HT-29d cells produced stronger PI3-kinase expression as compared with the non-anetastatic WiDr cells and the low metastatic HT-29 cells in vivo. Conclusion: The selected variant cell lines can exhibit an enhanced metastatic potential. The level of uPA and PAI- 1 are positively correlated with the metastatic capacity of tumor cells. The expression of PI3-kinasecorrelates with tumor development and metastasis.