摘要
目的从高转移潜能的人肝癌细胞系中分离出不同转移潜能的单克隆细胞株。方法采用有限稀释法,对高转移潜能人肝细胞癌细胞系 MHCC97进行单克隆培养,筛选不同转移潜能的克隆细胞株,测定目标克隆株的生长速度、核型、AFP 分泌情况、体外侵袭能力、裸鼠接种肺转移率。结果筛选出高、低两个不同转移潜能的单克隆细胞株,分别命名为 MHCC97-H 和 MHCC97-L,前者肺转移率为100%,后者为40%;MHCC97-H 的细胞倍增时间为34.2h,MHCC97-L为60.0h;流式细胞分析显示,MHCC97-H 与 MHCC97-L 相比,前者 G_0-G_1期细胞比例低,S 期、G_2-M 期细胞比例高;人工基底膜穿透能力前者明显强于后者。结论本实验证实了 MHCC97癌细胞系的异质性。这两个单克隆细胞株对于肝癌生物学行为的比较研究有一定的使用价值。
Objective To establish hepatocellular carcinoma(HCC)cell clones with various me- tastatic potentials from highly metastatic HCC cell line MHCC97.Methods Cell clone culture was conducted on the 25th passage of the source cell line MHCC97 using limited dilution method.Target cell clones were selected by in vivo screening in nude mice.Biological characteristics of the target clones were studied in terms of tumor cell doubling time,karyotyping,in vitro invasiveness,alpha-fe- toprotein secretion and pulmonary metastasis after orthotopic implantation.Results HCC clones with high(MHCC97-H)and low(MHCC97-L)metastatic potentials were established from the source cell line MHCC97.When MHCC97-H was compared with MHCC97-L,the pulmonary metastatic rate was 100%(10/10)versus 40%(4/10),tumor cell doubling time 34.2 hours vs.60.0 hours,and Boyden chamber in vitro invasion assay was 37.5±11.0 cells/field vs.17.74±6.3 cells/field.Karyotyping re- vealed that the main image of chromosomes was 55—57(68%)in MHCC97-H and 57—62(58%)in MHCC97-L.The proportions of cells in G_0—G_1 phase,S phase,and G_2-M phase for MHCC97-H/ MHCC97-L were 56.10%/65.32%, 28.35%/24.96% and 15.64%/9.73%,respectively,as meas- ured by flow cytometry.The serum AFP levels in nude mice in 5 weeks after orthotopic implantation of tumor tissue were 245.8±66.1μg/L for MHCC97-H and 91.2±65.7μg/L for MHCC97-L.Con- clusions The results confirm the heterogeneity of MHCC97.These tow clones of the same genetic background are of great value for further investigation on metastasis of HCC.
出处
《中华肝胆外科杂志》
CAS
CSCD
2001年第11期681-685,共5页
Chinese Journal of Hepatobiliary Surgery
基金
国家重点基础研究发展规划"恶性肿瘤发生与发展的基础性研究"G1998051211
上海市医学领先专业重大项目:"肝癌转移的分子机制及干预治疗的实验研究"983001
