摘要
目的:探讨人结直肠癌肿瘤干样细胞的生物特性及其与自噬发生的关系。方法:采用无血清培养法从人结直肠癌HCT116细胞中获取HCT116球细胞(结直肠癌肿瘤干样细胞);分别用克隆集落形成实验、成球实验、Transwell实验检测HCT116细胞与HCT116球细胞的增殖能力、自我更新能力及侵袭和迁移能力,用免疫荧光法检测两种细胞中自噬标志物LC3B的荧光表达,并用RT-PCR和Western blot分别检测两种细胞中LC3B及自噬相关基因ATG5、ATG7的mRNA与蛋白表达。结果:HCT116球细胞克隆形成能力、新成球能力以及侵袭与迁移能力均较HCT116细胞明显增强(均P<0.05)。HCT116球细胞中LC3B的荧光表达强度较HCT116细胞明显增高(P<0.05);HCT116球细胞中LC3B、ATG5、ATG7的mRNA和蛋白表达量较HCT116细胞均显明显升高(均P<0.05)。结论:结直肠癌肿瘤干样细胞较结直肠癌细胞具有更强的自我更新和体外增殖能力,以及更强的侵袭和迁移能力,该生物学特性可能与其高自噬活性密切相关。
Objective: To investigate the biological characteristics of human colorectal cancer stem-like cells and their relationship with autophagy. Methods: HCT116 sphere cells(colorectal cancer stem-like cells) were dissociated from the human colorectal cancer HCT116 cells by culture with serum-free medium. The proliferative and self-renewal abilities as well as the invasion and migration abilities of HCT116 cells and HCT116 sphere cells were detected by colony formation assay, sphere-forming assay, and Transwell assay respectively. In addition, the fluorescence expressions of the autophagy marker LC3 B in the two types of cells were detected by immunofluorescence assay, and the mRNA and protein expressions of LC3 B and autophagy-associated genes ATG5 and ATG7 in the two types of cells were determined by RT-PCR and Western blot respectively.Results: The colony formation and sphere formation abilities as well as invasion and migration abilities of HCT116 sphere cells were significantly stronger than those in HCT116 cells(all P<0.05). The fluorescence intensity of LC3 B in HCT116 sphere cells was significantly enhanced compared with HCT116 cells(P<0.05). The mRNA and protein expressions of LC3 B, ATG5 and ATG7 in HCT116 sphere cells were significantly higher than those in HCT116 cells(all P<0.05).Conclusion: Colorectal cancer stem-like cells have enhanced self-renewal and proliferative abilities as well as increased invasion and migration abilities in vitro compared to colorectal cancer cells, and these biological characteristics may be closely related to their autophagic activities.
作者
侯松林
蒋先虹
周国俊
李利发
张广军
周彤
HOU Songlin;JIANG Xianhong;ZHOU Guojun;LI Lifa;ZHANG Guangjun;ZHOU Tong(The Second Department of Gastrointestinal Surgery,Affiliated Hospital of North Sichuan Medical College,Nanchong,Sichuan 637000,China;Institute of Hepato-Biliary-Pancreas and Intestinal Disease.North Sichuan Medical College,Nanchong,Sichuan 637000,China)
出处
《中国普通外科杂志》
CAS
CSCD
北大核心
2019年第10期1253-1260,共8页
China Journal of General Surgery
基金
川北医学院科研发展计划资助项目(CBY17-A-ZD06)
四川省南充市市校联合合作科研专项资金(18SXHZ0548
18SXHZ0511)
关键词
结直肠肿瘤
肿瘤干细胞
自噬
肿瘤侵润
Colorectal Neoplasms
Neoplastic Stem Cells
Autophagy
Neoplasm Invasiveness