摘要
背景:白细胞介素8是一种重要的炎性趋化因子,其在调节肿瘤细胞增殖、血管新生方面发挥了重要作用。目的:探讨白细胞介素8对CD133^+肝癌细胞侵袭转移能力的影响。方法:分离培养人肝癌MHCC97-H细胞株,免疫磁珠分选出CD133^+/CD133-MHCC97-H细胞,流式细胞仪检测CD133表达,ELISA检测上清液白细胞介素8水平。克隆形成实验、裸鼠成瘤实验、Transwell小室分别检测CD133^+/CD133-MHCC97-H细胞克隆形成率、小鼠成瘤能力、细胞迁移侵袭能力。另外,采用白细胞介素8中和抗体处理细胞,比较CD133^+/CD133-MHCC97-H细胞CD133表达、上清液中白细胞介素8水平及细胞克隆形成能力和迁移、侵袭能力。结果与结论:(1)CD133^+MHCC97-H细胞的CD133表达率、上清液白细胞介素8水平及克隆形成率均显著大于CD133-MHCC97-H细胞(P<0.05);(2)细胞浓度为1×10~6 L^(-1)和1×10~7 L^(-1)的CD133^+MHCC97-H细胞接种后有部分小鼠出现皮下移植瘤,但CD133-MHCC97-H细胞在同样细胞浓度下,均未出现皮下移植瘤;(3)CD133^+MHCC97-H细胞的透膜细胞数量均显著大于CD133-MHCC97-H细胞(P<0.05);(4)经白细胞介素8中和抗体处理之后,CD133^+/CD133-MHCC97-H细胞的CD133表达率、上清液中白细胞介素8水平、克隆形成率均显著下降(P<0.05),且CD133^+MHCC97-H细胞下降程度更大(P<0.01);CD133^+MHCC97-H细胞的迁移、侵袭透膜细胞数显著减少(P<0.05),CD133-MHCC97-H细胞则未出现明显的改变(P>0.05);(5)结果表明,白细胞介素8可以特异性参与调节CD133^+MHCC97-H细胞侵袭、转移能力。
BACKGROUND： Interleukin-8 is an important inflammatory chemokine that plays an important role in the regulation of tumor cell proliferation and angiogenesis. OBJECTIVE： To investigate the effect of interleukin-8 on the invasion and metastasis of CD133＋ hepatocellular carcinoma stem cells. METHODS： After isolation and culture of MHCC97-H cell lines, CD133＋/CD133- MHCC97-H cells were sorted using immunomagnetic beads. CD133 expression was detected using flow cytometry, and interleukin-8 level in supernatant was measured using ELISA method. Cloning efficiency, tumorigenic capacity, cell migration and invasion ability were detected through colony formation assay, tumorigenesis experiment in nude mice, and Transwell detection. Additionally, other cells were neutralized using interleukin-8 neutralizing antibody. Measurement results were compared between cells undergoing different treatments. RESULTS AND CONCLUSION： The CD133 level, interleukin-8 level, cloning efficiency and cell membrane permeability of CD133＋MHCC97-H cells were significantly higher than those of CD133-MHCC97-H cells（P〈0.05）. Transplantation of CD133＋MHCC97-H cells at 1×10^6/L and 1×10^7/L resulted in subcutaneous tumors in some mice, whereas no subcutaneous tumors appeared in mice undergoing transplantation of CD133-MHCC97-H cells at the same concentrations. After interleukin-8 neutralizing antibody treatment, the CD133 level, interleukin-8 level, and cloning efficiency of CD133＋/CD133-MHCC97-H cells were significantly decreased（P〈0.05）, especially in the CD133＋MHCC97-H cells（P〈0.01）; the migration and invasion ability and cell membrane permeability of CD133＋MHCC97-H cells were significantly reduced（P〈0.05）, but these changes were not obvious in CD133-MHCC97-H cells（P〉0.05）. These results show that interleukin-8 could be specifically involved in the invasion and metastasis of CD133＋MHCC97-H cells.
出处
《中国组织工程研究》
CAS
北大核心
2016年第41期6145-6150,共6页
Chinese Journal of Tissue Engineering Research
关键词
干细胞
肿瘤干细胞
肝癌
CD133
细胞侵袭
细胞转移
,Liver Neoplasms
Neoplastic Stem Cells
Antigens, CD
Neoplasm Invasiveness
Neoplasm Metastasis
Tissue Engineering
