CD90对肝癌细胞侵袭及上皮间质转化的影响

The effect of CD90 on the hepatocellular carcinoma invasion and EMT transformation

目的分析乙型病毒性肝癌患者肝癌组织和乙型病毒性肝癌细胞株中侵袭相关基因和EMT相关基因表达情况并探讨其与CD90表达的相关性。方法采用细胞迁移实验分析细胞迁移能力,通过RT PCR法分析Hep G2、Hep3B、Huh7、PLC/RFP/6、SNU423、SNU449和SNU475等肝癌细胞系中的CD90、侵袭相关基因MMP1、MMP2和EMT调控剂E-cadherin、N-cadherin的m RNA表达水平。同样对病毒性乙型肝炎引起肝癌中49个样本进行了相同基因检测以及病理特征分析。结果在所有的肝癌细胞系中,CD90阳性细胞比阴性细胞的细胞迁移性活跃,上调侵袭性和上皮细胞-间充质转化的相关基因中N-钙黏素的表达增加,而E-钙黏蛋白表达降低。该实验将肝癌细胞按CD90 m RNA的含量分成高CD90组(CD90含量最高的40%标本)和低CD90组(CD90含量最低的40%标本)。同低CD90组相比,高CD90组的血管侵犯频繁,侵袭性相关基因MMP1(P=0.01)、MMP2(P=0.003)和EMT调节因子E-cadherin(P=0.022)的表达含量高。结论 HCC中CD90高表达,CD90高表达肝癌细胞能通过上调侵袭性相关基因和EMT相关基因而产生更强的侵袭性。

【Objective】To analyze invasion and EMT-associated genes expression in hepatocellular carcinoma tissue and liver cancer cell lines with high expression of CD90. 【Methods】The m RNA expression levels of CD90,invasion-associated genes MMP1 and MMP2, and EMT regulators（E-cadherin and N-cadherin） were examined by real-time PCR in HCC cell lines including Hep G2, Hep3 B, Huh7, PLC/RFP/6, SNU423, SNU449, and SNU475.Same genes and pathological features were also investigated in 49 samples of hepatitis B virus-related human HCCs.【Results】In all HCC cell lines studied, CD90-positive cells showed higher cell migration activity and up-regulated invasion- and EMT-associated genes with increased N-cadherin and decreased E-cadherin expressions compared to CD90-negative cells. The human HCCs were divided into the CD90-high group（top 40%） and the CD90-low group（bottom 40%） according to the level of CD90 m RNA. The CD90-high group showed relatively frequent vascular invasion and significantly higher expression of invasion-associated genes [MMP1（P =0.01） and MMP2（P =0.003）] and EMT regulators [E-cadherin（P =0.022）] compared to the CD90-low group.【Conclusion】Our results suggest that there is a subtype of HCC with high expression of CD90, which might have more invasive characteristics by up-regulation of invasion-associated genes and EMT-associated genes.