肾母细胞瘤中ABCG2的表达及侧群的检测

Expression of ABCG2 in Wilms＇ tumor and their detection in side-population

目的检测肾母细胞瘤中是否存在肿瘤干细胞标记物ABCG2的表达，同时应用拒染方法检测该肿瘤中是否存在侧群（sidepopulation，SP）细胞。方法①收集2009年1月至2011年5月34例肾母细胞瘤手术标本，用免疫组织化学S-P方法（链霉菌抗生物素蛋白一过氧化物酶连结法）检测肿瘤干细胞标记物ABCG2的表达。②收集6例手术新鲜组织标本进行原代培养，制取单细胞悬液，进行荧光染料Hoechst33342染色，用荧光显微镜观察是否含有拒染的细胞；并使用流式细胞仪检测侧群的表达。结果肾母细胞瘤中存在ABCG2的表达，阳性率约为76．4％（26／34）；6例新鲜组织中3例经染色后在荧光显微镜下发现少量染色浅的拒染细胞；流式细胞仪检测到原代肾母细胞瘤中含有少量可被维拉帕米阻断的拒染细胞，其表达量约为0．7％。结论肾母细胞瘤中存在AB—CG2的表达，可能与肿瘤耐药相关；应用拒染方法可以测得原代肾母细胞瘤中含有小群SP细胞，它们中可能含有肿瘤干细胞。

Objective To explore the expression of stem cell marker ABCG2 in Wilms＇ tumor, and used flow cytometer to detect whether the side population（ SP ） could found in them. Methods Totally 34 cases of Wilms＇ tumor specimens were collected from January 2009 to May 2011; the expression of tumor stem cell marker ABCG2 was detected via immunohistochemistry assay. Furthermore, 6 specimens were collected for single cell suspension, followed hy staining with fluorescent dye Hoechst 33342 and observation using fluorescence microscopy. Additionally, flow Cytometer was conducted to investigate the side-population. Results About 76. 4% （26 / 34） of specimens could be found the expression of ABCG2. In 3/6 cases, we observed a small amounts of staining after Hoechst 33342 dyeing; 3, and appropriately 0. 7% of SP cells were detected by FCS, which could be blocked by verapamil. Conclusions The expression of ABCG2 in Wilms＇ tumor may correlate to tumor multi-drug resistance; A small number of SP cells were detected in WT, in which may contain cancer stem cells.