人肝细胞癌细胞亚群的克隆分离及异质性机制的初步研究  

Elementary study on subpopulation cell isolation and tumor heterogeneity of human hepatocellular carcinoma

作　　者：颜政[1] 方驰华[1] 

机构地区：[1]南方医科大学珠江医院普外科,广东省广州市510282

出　　处：《世界华人消化杂志》2006年第5期481-485,共5页World Chinese Journal of Digestology

基　　金：广东省自然科学基金资助课题;No.2002-020097~~

摘　　要：目的:通过原代培养人肝癌细胞、克隆分离异质性亚群细胞,探讨肝癌异质性机制．方法:原代培养人肝细胞癌细胞,应用有限稀释法对培养的肝癌细胞进行单细胞克隆分离异质性亚群,并应用细胞计数法测定其细胞倍增时间及倍增数,流式细胞仪检测其DNA含量确定细胞周期分布,裸鼠异体移植检测其成瘤能力．结果:分离到LCSC-1及LCSC-2两个细胞亚群．LCSC-1亚群细胞呈长梭形,裸鼠异体移植不能成瘤(0／8);LCSC-2亚群细胞呈多角形、多突起,裸鼠异体移植可成瘤(8／8)．LCSC-1与 LCSC-2相比,体外增殖能力强,倍增时间(18．6 ±3．2 h vs 25．9±4．7 h)和最大倍增倍数(16．1 ±1.4 vs 12．2±1．6)有显著差异(均P<0．01);而且LCSC-1处于细胞周期S(28．4％±3．3％vs 20．2％±1．9％,P<0．01)和G2M(25．0％±6．3％ vs 16．6％±4．7％,P<0．05)的比例明显高于 LCSC-2．结论:原发性肝细胞癌中存在着异质性的肿瘤细胞亚群,可能来源于肿瘤干细胞的分化．AIM： To isolate and characterize the heterogeneous subpopulation cells from human hepatocellular carcinoma （HCC）, and to explore the mechanism of HCC heterogeneity. METHODS： Tumor samples were obtained from 1 patient with HCC after surgery, and then HCC cells were cultured in vitro by primary tissue culture technique. Heterogeneous subpopulations were isolated by limiting dilution and their growths were observed with the help of cell counts. The cycle and DNA content of tumor cells were investigated with flow cytometry. Each subpopulation was implanted in nude mice subcutaneously to measure the tumor forming ability. RESULTS： Two subpopulations of HCC cells, LCSC-1 and LCSC-2, were derived from primary tumor tissues. LCSC-1 cells show a sharp and spindle feature, and could not form new tumor when implanted in nude mice （0/8）. However, LCSC-2 cells were multangular in shape, with protuberances, and could produce new tumor （8/8）. In comparison with LCSC-2 cells, LCSC-1 had stronger proliferation ability, and the double time （18.6 ± 3.2 h vs 25.9 ± 4.7 h） and maximum multiples （16.1 ± 1.4 vs 12.2 ± 1.6） were markedly different between the two （P 〈 0.01）. Furthermore, the ratio of cells in S and G2M phase was significantly higher for LCSC-1 than those for LCSC-2 （S phase： 28.4% ± 3.3% vs 20.2% ± 1.9%, P 〈 0.01; G2M phase： 25.0% ± 6.3% vs 16.6% ± 4.7%, P 〈 0.05）. CONCLUSION： HCC cells are heterogeneous with respect to characteristics on morphology, proliferation and tumorigenesis potentiality. The heterogeneity of HCC may arise from the differentiation of tumor stem cells.

关 键 词：肝癌 肿瘤异质性 肿瘤干细胞 

分 类 号：R735.7[医药卫生—肿瘤]