结直肠癌CD133+细胞定量评估的意义

目的研究CD133+结直肠癌干细胞与肿瘤增殖、转移的相关性。方法取得术后新鲜的结直肠癌组织后,立刻进行清洗、消化、培养等过程,得到了具有活性的原代结直肠癌单细胞。特异性抗体CD133标记后运用流式细胞技术检测癌干细胞分布比例。癌组织同时进行免疫组化和分子生物学研究,确定增殖蛋白Ki-67、抑制肿瘤转移相关蛋白e-cadherin和细胞凋亡相关蛋白caspase-3的表达。结果结直肠癌原代细胞分离、纯化和检测技术简明可行,所得数据客观准确。研究病例按照CD133+癌干细胞比例≥3%和<3%分成两组。结果显示CD133+癌干细胞≥3%组在肿瘤大小和淋巴转移有增大和增多趋势;肿瘤增殖特异性蛋白Ki-67增高具有显著性差异;抑制肿瘤转移相关蛋白e-cadherin的表达降低;凋亡相关蛋白caspase-3的表达降低。结论本研究成功建立了一套可行性高的结直肠癌CD133+干细胞准确定量评估系统。结直肠癌干细胞的准确定量检测可以作为评估患者预后和化疗敏感度的一项重要指标。本技术也为进一步提纯结直肠癌干细胞,并且最终研究对此靶点的攻击提供了平台。

Importance of Quantitative Evaluation of Colorectal Cancer CD133~+ Cells

Objective To precisely quantify the percentage of CD133+ cancer stem cells by isolation and purification of primary colorectal cancer cells,and to clarify its relationship with cancer proliferation and metastasis.Methods Primary colorectal cancer cells were obtained after washing,digesting and culturing fresh colorectal cancer tissues.Cancer stem cells were labeled with specific CD133 antibody and analyzed by flow cytometry.Cancer tissues were also tested using immunostaining and Western blotting to detect expression of proliferative marker Ki-67,invasion suppressor related protein e-cadherin,and apoptosis related protein caspase-3.Results Isolation,purification and precisely quantification of primary colorectal cancer stem cells were efficiently modified as feasible procedures.Patients were divided into two groups with percentage of CD133+ cancer stem cells ≥3% or<3%.In the ≥3% group,a increase of tumor size and lymphatic metastasis was observed,expression of the specific proliferation protein Ki-67 was significantly increased,production of the tumor metastasis suppressor protein e-cadherin and the apoptotic protein caspase-3 was decreased.Conclusion A precisely quantitative evaluation system of the primary colorectal cancer stem cells was feasibly established in our study.The system can be used as indicators of prognosis and sensibility to chemotherapy in colorectal cancer patients.The technique also functions as a platform in purification of colorectal cancer stem cells and cancer-stem-cell-targeted therapy in the future.