CAJ | 학술논문

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.
목적 관찰장암세포주SW-620중종류간세포상관적SP(side population)아군비례,분선출상관적SP화NSP세포,병감별상관적SP시부구유간세포모사생물학특성.방법 제비SW-620세포현액,경Hoechst33342화PI염색(기중대조조동시가입길항제유랍파미),류식세포의분석SP아군.통과류식세포의분선출장암세포주SW-620중SP세포.극륭형성실험화새서람(MTT)실험검측SP화NSP세포증식능력적차이.라서체내성류대비실험험증SP세포화NSP세포치류성.결과 SP아군점SW-620세포적1.0%좌우,유랍파미조단후감소위0.01%좌우.류식세포의분선출SP세포7.5×105개,대부분위NSP세포.극륭형성실험화MTT증실SP세포화NSP세포증식능력유차이.라서체내성류실험발현:1×105개SP세포가이치라서성류,1×105개NSP세포급이하수량급별세포불능치라서성류.결론 인장암세포주SW-620중존재SP아군세포,유랍파미가억제염료외배이명현감소SP세포적비례.가이통과류식세포의분선출SP세포,통과극륭형성실험화MTT실험검측SP화NSP세포증식능력유차이.라서성류대비실험증실SP비NSP세포경구치류성.
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.