肝癌干细胞CD90+和ESA+亚群生物学特性研究

Biological features of CD90+ and ESA+ liver cancer stem cell

关键词:  肝癌  肿瘤干细胞  CD90  ESA

Key words:  liver cancer  cancer stem cell  ESA  CD90

摘要:

目的　研究肝癌干细胞不同亚群CD90+和ESA+细胞的生物学特性，为肝癌的治疗及预后判定提供新的思路。方法　采用活细胞流式细胞术检测肝癌细胞系MHCC97L中肝癌干细胞标志物CD90、ESA的表达情况，并分选ESA+，CD90+细胞，通过甲基纤维素成球实验、Transwell、CCK8法进行体外生物学特征研究。结果　肝癌细胞系MHCC97L中CD90+和ESA+细胞表达比例分别为2.37%和3.70%。ESA+细胞成球率明显高于ESA-细胞［成球率（35.6±2.1）% vs.（9.6±1.4）%, P<0.01］，CD90+细胞成球率明显高于CD90-细胞［成球率（29.2±1.3）% vs.（7.4±0.6）%］（P<0.01），ESA+细胞成球率明显高于CD90+细胞且ESA+细胞sphere球体积明显大于CD90+细胞体积；ESA+细胞较ESA-细胞侵袭能力提高2.12倍［（282±15.1 vs. 133±12.4），P<0.01］，CD90+细胞较CD90-细胞侵袭能力提高2.04倍［（231±18.1 vs. 113±10.2），P<0.01］。CD90+细胞耐药性明显强于ESA+细胞［IC50（0.98 μg/mL vs. 0.36 μg/mL）］，ESA+细胞增殖能力明显强于CD90+细胞。结论　肝癌组织中存在不同的干细胞亚群ESA+细胞及CD90+细胞，这些亚群具有不同的生物学特性，影响肝癌的耐药、侵袭，了解肝癌中不同干细胞亚群的表达情况可用于指导临床个体化治疗。

Abstract:

Objective To study the biological features of CD90+ and ESA+ Liver Cancer Stem cell and provide a new view for treatment and prognosis of liver cancer. Methods Flow cytometry was used to detect and sort CD90+ and ESA+ cells in MHCC97L cell line. Colony formation, drug resistance, invasion and proliferation experiment were tested in those cells. Results CD90+ and ESA+ cells was 2.37% and 3.70%, respectively in MHCC97L cell line. Colony formation rate in ESA+ cell was higher than in ESA- cell ［(35.6±2.1)% vs. （9.6±1.4)%, P<0.01］, and that in CD90+ cell was higher than in CD90- cell ［(29.2±1.3)% vs.(7.4±0.6)%, P<0.01］. Colony formation rate and the cell sphere volume of ESA+ was higher than CD90+ cell. Invasion ability of ESA+ and CD90+ cell improved 2.12 and 2.04 times respectively, compared with ESA- and CD90- cell. Cisplatin resistance ability of CD90+ cell was higher than ESA+cell (IC50: 0.98 μg/mL vs.0.36 μg/mL）. Conclusion Different sub-population stem cell of liver cancer would show variety invasion, proliferation ability and sensitivity to drugs, which would affect individualized treatment.