【摘要】 目的探讨CD200在大肠癌干细胞中的表达及其对患者细胞免疫水平的相关性。方法选取2015年4月至2017年5月广东省广州市花都区人民医院收治的大肠癌患者58例作为对象,所有患者均行手术治疗,取大肠癌病灶组织,采用差速离心法完成大肠癌干细胞分离、培养。根据手术效果将患者分为完全缓解组(42例,病灶切除,近6个月未见复发)和未缓解组(16例,病灶切除但出现复发或病灶未能全部切除,病情持续发展)。采用流式细胞仪测定两组样本中CD200阳性细胞数;采用酶联免疫吸附试验测定两组白细胞介素-17 (IL-17)、白细胞介素-10 (IL-10)和转化生长因子β1 (TGF-β1);采用SPSS Pearson?相关性分析软件对CD200在大肠癌中与肿瘤细胞免疫水平相关性进行分析。结果倒置显微镜下结肠癌干细胞接种在胎牛血清10. 0%培养基中,第3天仍可见干细胞球,多数细胞悬浮生长,部分细胞从细胞求中逸出呈梭形;第7天多数干细胞贴壁生长,呈梭形;第9天干细胞全部转化为梭状大肠癌干细胞,且贴壁生长,流式细胞仪测定结果表明:结肠癌干细胞表面标志物CD200细胞群含量达到80. 3%;大肠癌患者中完全缓解组CD200表达水平低于未缓解组(P <0. 01);完全缓解组IL-17、INF-γ、IL-10及TGF-β1水平,均高于未缓解组(P <0. 01);完全缓解组IL-4水平,低于未缓解组(P <0. 01)。大肠癌干细胞CD200表达水平与FoxP3~+/CD4~+、IL-4呈正相关(P <0. 05);大肠癌干细胞CD200表达水平与IL-17、INF-r、IL-10及TGF-β1水平呈负相关(P <0. 05)。结论 CD200在大肠癌干细胞中呈高表达,且与肿瘤患者细胞免疫水平具有一定的相关性,加强CD200表达水平测定能评估、预测疾病严重程度,指导临床治疗,值得推广应用。更多还原

【Abstract】 Objective To investigate the expression of CD200 in colorectal cancer stem cells and its correlation with cellular immune levels. Methods A total of 58 patients with colorectal cancer admitted to the Department of Gastroenterology,Huadu District People’s Hospital,Guangzhou City,Guangdong Province,from April 2015 to May 2017 were enrolled. All patients underwent surgery for colorectal cancer lesions using differential centrifugation. The method completes the isolation and culture of colorectal cancer stem cells. According to the surgical results,the patients were divided into complete remission group( n = 42 cases,lesion resection,no recurrence in the past 6 months) and non-remission group( n = 16 cases,resection of the lesion but recurrence or failure of the lesions were completely removed,the condition Continuous development). The number of CD200 positive cells in the two groups was determined by flow cytometry; two groups of interleukin-17( IL-17),interleukin-10( IL-10) and transforming growth factor β1 were determined by enzyme-linked immunosorbent assay.( TGF-β1); SPSS Pearson correlation analysis software was used to analyze the correlation between CD200 and tumor cell immunity in colorectal cancer. Results Under the inverted microscope,colon cancer stem cells were inoculated in fetal bovine serum 10. 0% medium. Stem cell spheres were still visible on the third day. Most of the cells were suspended and some cells escaped from the cells to form a fusiform shape. On the 7 th day,most stem cells adhered. Growth,spindleshaped; the 9 th stem cells were all transformed into spindle-shaped colorectal cancer stem cells,and adherent growth,flow cytometry results showed that the colon cancer stem cell surface marker CD200 cell population content reached 80. 3%; colorectal cancer patients completely The expression level of CD200 in the remission group was lower than that in the non-remission group( P < 0. 05). The levels of IL-17,INF-γ,IL-10 and TGF-β1 in the complete remission group were higher than those in the non-remission group( P <0. 05); complete remission The level of IL-4 was lower in the group than in the non-remission group( P < 0. 05). The results of SPSS Pearson correlation analysis showed that CD200 expression in colorectal cancer stem cells was positively correlated with Fox P3~+/CD4~+and IL-4( P < 0. 05); CD200 expression in colorectal cancer stem cells was related to IL-17,INF-γ,IL-10 and TGF-β1 levels were negatively correlated( P < 0. 05). Conclusion CD200 is highly expressed in colorectal cancer stem cells,and has a certain correlation with the cellular immune level of tumor patients. Strengthening the expression of CD200 can evaluate and predict the severity of the disease and guide clinical treatment. It is worthy of popularization and application.更多还原