摘要
目的建立肝癌裸鼠皮下移植瘤动物模型,以癌干细胞的耐药特性为建立富集肝癌干细胞的切入点,在机体环境下进行构建体内肝癌干细胞的富集模式。方法将BEL-7404人肝癌细胞株的细胞悬液注射裸鼠两侧腋窝皮下,建立皮下移植瘤模型,经阿霉素干预后原代取材,分别以单个细胞克隆形成实验、平板克隆形成实验和悬浮球形成实验,对亲本细胞及经阿霉素干预后的BEL-7404肝癌细胞进行癌干细胞相关细胞学特性的检测。结果每一代裸鼠皮下移植瘤模型成瘤率为100%,亲本细胞与经体内阿霉素干预后的第一代BEL-7404肝癌细胞的单细胞克隆形成率比较没有明显差异(P>0.05),而体内干预第二代细胞的单细胞克隆形成率均明显高于亲本细胞和经干预后的第一代细胞(P<0.05),体内干预第一代细胞平板克隆形成率均明显低于亲本细胞和第二代细胞(P<0.05)。亲本细胞及经体内阿霉素干预后的BEL-7404肝癌细胞在干细胞培养基中培养均未见有细胞悬浮球形成。结论经低剂量阿霉素体内干预可作为富集肝癌干细胞的手段之一。
Objective To establish a nude mouse model of liver cancer and explore whether drug resistance of cancer stem-like ceils in vivo can lead to enrichment of stem-like cells. Methods A subcutaneous tumor model of liver carcinoma was established by subcu- taneously injecting human liver BEL-7404 cell suspensions into both axilla of BALB/C-nu mice.Following adriamycin treatment, pri- mary cultures were established, and both parental and adriamycin-treated cells were studied in assays of single-cell colony formation and sphere cell formation to detect drug resistance and analyze other cytological characteristics. Results The tumor formation rate in this BALB/C-nu subcutaneous tumor model was 100%.There were no significant differences in the rate of single-cell colony formation between BEL-7404 parental liver cells and first-generation cells after adriamycin treatment（P〉0.05 ）.However, second-generation cells showed a significantly higher rate of single-cell colony formation and a significantly lower colony formation rate than parental or first- generation cells （P〈0.05）. Non-adherent spheres were not observed in cultures of any of the three cell types. Conclusions Low-dose adriamycin intervention may be a way to enrich for liver cancer stem-like cells in vivo.
出处
《中国癌症防治杂志》
CAS
2013年第2期92-95,共4页
CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT
基金
国家自然科学基金资助项目(30860326)
广西自然科学基金资助项目(0991139)
广西科技创新能力与条件建设项目(桂科攻0993003C-1)
关键词
肝肿瘤
癌干细胞
皮下移植瘤模型
阿霉素
Liver neoplasms
Cancer stem cell
Subcutaneous tumor model
Adriamycin
