肠癌SW-620肿瘤细胞SP亚群研究

目的 观察肠癌细胞株SW-620中肿瘤干细胞相关的SP(side population)亚群比例,分选出相关的SP和NSP细胞,并鉴别相关的SP是否具有干细胞某些生物学特性.方法 制备SW-620细胞悬液,经Hoechst33342和PI染色(其中对照组同时加入拮抗剂维拉帕米),流式细胞仪分析SP亚群.通过流式细胞仪分选出肠癌细胞株SW-620中SP细胞.克隆形成实验和噻唑蓝(MTT)实验检测SP和NSP细胞增殖能力的差异.裸鼠体内成瘤对比实验验证SP细胞和NSP细胞致瘤性.结果 SP亚群占SW-620细胞的1.0%左右,维拉帕米阻断后减少为0.01%左右.流式细胞仪分选出SP细胞7.5×105个,大部分为NSP细胞.克隆形成实验和MTT证实SP细胞和NSP细胞增殖能力有差异.裸鼠体内成瘤实验发现:1×105个SP细胞可以致裸鼠成瘤,1×105个NSP细胞及以下数量级别细胞不能致裸鼠成瘤.结论 人肠癌细胞株SW-620中存在SP亚群细胞,维拉帕米可抑制染料外排而明显减少SP细胞的比例.可以通过流式细胞仪分选出SP细胞,通过克隆形成实验和MTT实验检测SP和NSP细胞增殖能力有差异.裸鼠成瘤对比实验证实SP比NSP细胞更具致瘤性.

Side population in human colon cancer cell line SW-620

Objective To identify the ratio of the side population (SP) of cancer stem-like cells in colon cancer cell line SW-620 and separate the SP and NSP cells, and initially identify whether the SP cells have some stem-cell bionomics. Methods SW-620 tumor cells suspension was stained with Ho-echst33342 and PI in the absence or presence of verapamil. Then the SP was analyzed by flow cytometry ( FCM). The SP and NSP cells were separated by FCM. The disparity of SP and NSP cells was detected by colon forming and cell growth plot experiment. The oncogenicity of SP cells and NSP cells was identified in nude mice. Results SP cells accounted for 0.1%-1.0% of the SW-620 cells, which were decreased to 0.01% by verapamil. 7.5 × 105 SP cells were separated by FCM. SP and NSP cells had disparity in colon forming and cell growth plot experiment. The nude mouse oncogenicity experiment revealed that the nude mice could erupt tumor by injection of 1 × 105 SP cells, but the nude mice could not erupt tumor by injection of 1 × 105 NSP cells. Conclusion Human colon cancer cell line SW-620 contains SP that can be decreased by verapamil. SP cells can be separated by FCM. SP and NSP cells have disparity in colon forming and cell growth plot experiment. The nude mouse oncogenicity experiment reveals that SP cells have a higher oncogenicity than NSP cells.