人结肠直肠癌组织中侧群细胞分选、鉴定及其化疗耐药特性的初步研究  

Preliminary study on sorting,identification and chemoresistance of side population cells in human colorectal cancer tissues

作　　者：刘贵远[1] 周海华[1,2] 张超逸 徐洁 LIU Guiyuan;ZHOU Haihua;ZHANG Chaoyi;XU Jie(Taizhou People′s Hospital,Taizhou,Jiangsu,225300;Medical College of Yangzhou University,Yangzhou,Jiangsu,225001)

机构地区：[1]江苏省泰州市人民医院,江苏泰州225300 [2]扬州大学医学院,江苏扬州225001

出　　处：《实用临床医药杂志》2020年第4期31-35,共5页Journal of Clinical Medicine in Practice

摘　　要：目的分选人结肠直肠癌组织中侧群(SP)细胞,探讨其相关生物学特性。方法利用Hoechst33342染色荧光激活细胞分选(FACS)技术分选SP细胞,非侧群(NSP)细胞作为对照。三维(TD)培养分选的SP细胞,分析其在裸鼠体内致瘤能力。无血清培养观察SP细胞与NSP细胞的成球能力,水溶性四唑盐试剂(WST)分析SP细胞与NSP细胞的耐药性差异。结果人结肠直肠癌组织中SP细胞的比例为(0.90±0.10)%。SP细胞在无血清培养基中形成悬浮细胞球的能力强于NSP细胞。在103、104、105、106个SP细胞接种裸鼠皮下时,其成瘤率分别为30.0%、50.0%、70.0%、100.0%,而NSP细胞的成瘤率分别为0%、0%、40.0%、60.0%,差异有统计学意义(P<0.05)。耐药实验中,SP细胞对5-氟尿嘧啶(5-FU)的耐药性显著高于NSP细胞(P<0.05)。结论人结肠直肠癌组织中有SP细胞,SP细胞具有干细胞特性,SP细胞能在无血清培养基中以细胞球的形态存在,SP细胞对5-FU有耐药性。Objective To sort side population(SP)cells in human colorectal cancer tissues and explore its related biological characteristics.Methods SP cells were selected by Hoechst33342 staining and fluorescence activated cell sorting(FACS)technoligy,and non-side population(SP)cells were used as control.Three dimensional(TD)cultures of SP cells were used to analyze their tumorigenic ability in nude mice.The spheroidizing abilities of SP cells and NSP cells were observed in serum-free culture,and the differences of drug resistance between SP cells and NSP cells were analyzed by water soluble tetrazolium salt reagent(WST).Results The proportion of SP cells in human colorectal cancer was(0.90±0.10)%.SP cells in serum-free medium had better ability to form suspension cells than NSP cells.When 103,104,105 and 106 SP cells were inoculated subcutaneously in nude mice,the tumorigenesis rates were 30.0%,50.0%,70.0%and 100.0%respectively,while the tumorigenesis rates of NSP cells were 0%,0%,40.0%and 60.0%respectively,and there was significant difference in tumorigenesis rate between SP cells and NSP cells(P<0.05).The resistance rate of SP cells to 5-FU was significantly higher than that of NSP cells(P<0.05).Conclusions There are SP cells in human colorectal cancer tissue.SP cells have stem cell characteristics.SP cells can exist as cell spheres in serum-free medium.SP cells are resistant to 5-FU.

关 键 词：结肠直肠癌 侧群细胞 致瘤性 耐药性 分选 悬浮细胞球 抑制率 

分 类 号：R735.3[医药卫生—肿瘤]