人肝癌HepG2细胞异质性亚系的建立及生物学行为初探

A study on two heterogeneous sublines from HepG2 in their biological characteristics

目的:从人肝癌细胞株HepG2母系克隆分离异质性亚系,探讨肝癌异质性机理。方法:应用有限稀释法对培养人肝癌细胞株HepG2进行单细胞克隆分离异质性亚系,并应用细胞计数法、流式细胞仪检测其细胞周期和DNA含量,MTT法检测其对不同抗癌药物敏感性,同时应用裸鼠脾种植肝转移在体实验模型探讨其生物学行为的差异。结果:分离到HepG2-D3及HepG2-F4两个细胞亚系。HepG2-F4亚系细胞呈长梭形,体外增殖能力强,但脾脏内种植后不成瘤,不转移(0/10);HepG2-D3亚系细胞呈多角形、多突起,脾脏内种植成瘤率高(10/10),并在肝脏形成广泛转移(10/10)。结论:从人肝癌细胞株HepG2母系成功分离建立了两个异质性细胞亚系,两者在形态学、增殖能力及其对不同抗癌药物敏感性、成瘤和转移能力方面均存在差异,该亚系可为进一步研究肝癌异质性尤其是转移的确切机制提供良好的素材。

Aim: To isolate and characterize heterogeneous sublines from human hepatocellular (HepG2), explore the mechanism of HCC heterogeneity. Methods: Heterogeneous sublines were isolated with limited dilution from human hepatocellular carcinoma cell line (HepG2) and their growth was observered by counting cell in vitro. Tumor cell's period and DNA content were investigated and the effects of four anti-cancer drugs on these sublines were determined with the flow cytometry and MTT assay method respectively. The metastatic potential of these heterogeneous sublines in vivo was studied by constructing a liver-metastasizing nude mouse model from spleen inoculation.Results: The HepG2-D3 and HepG2-F4 sublines derived from carcinoma cell line HepG2 exhibited different characters: HepG2-F4 shows a sharp shape, with a high growth rate in vitro, but can not survive when transplanted in spleen (0/10). HepG2-D3 has a multi-angle shape, can survive and metastasize into liver diffusely (10/10).Conclusion:Two heterogeneous sublines derived from HepG2 have isolated and show different characters on morphology, proliferation, sensitivity to different anti-cancer drugs and metastatic potential, et al. Establishment of these two heterogeneous sublines may be valuable for further study on the molecular mechanisms of the heterogeneity and metastasis in hepatocellular carcinoma.