RNA干扰Oct4的表达对肝癌细胞生物学行为的影响

The effect of SiRNA -Oct4 expression on HCC cell line HepG2

【摘要】目的探讨干细胞相关基因Oct4与Wnt／β-catenin在人肝癌组织及肝癌细胞系中的相互作用。方法PCR法检测Oct4、Wnt／β-catenin及其他相关基因在肝癌组织、癌旁组织及正常肝组织的表达差异。使用SiRNA-Oct4沉默人肝癌HepG2细胞Oct4的表达，实时荧光定量PCR法检测Wnt／β-catenin基因表达。检测RNAi后细胞迁移以及克隆形成能力。结果肝癌患者的上述组织中，Oct4和β-catenin在癌内表达最高，癌旁次之，正常肝最低。使用SiRNA-Oct4沉默人肝癌HepG2细胞Oct4后，Oct4表达明显下调，β-catenin、Wntl0b表现为与Oct4表达呈正相关，TCF3表达与Oct4呈负相关。RNAi后HepG2细胞的迁移能力、克隆形成能力均下降。结论Oct4在肝癌组织内高表达而在正常肝细胞中的表达极低。RNAi后HepG2细胞的迁移能力、克隆形成能力均下降可能与Wnt／β-catenin通路功能减弱有关。

Objective To investigate the expression of Oct4 in liver cancer, and the interrelation of the Oct4 and Wnt/β-catenin genes in hepatocellular carcinoma（HCC） cell line HepG2. Methods RT- PCR technique was used to detect the expression of Oct4 and β-catenin in HCC specimens ; RNAi was used to knock-down the expression of Oct4 in HepG2, and the change of Wnt/β-cateninrelated genes were detected by Real time-PCR. Results In HCC specimens, the expression of Oct4 and β-catenin in tumor and cirrhotic liver tissues were stronger than normal liver tissues. In SiRNA Oct4 HepG2 cells, the expression of Oct4 was downregulated, and β-catenin as well as Wntl0b were in a positive correlation with Oct4, TCF3 was in negative correlation with Oct4. Clone formation and move ability of the HepG2 were downregulated. Conclusions The expression of Oct4 was higher in tumor tissues than in normal liver tissues. Silencing Oct4 by SiRNA-Oct4 in HepG2 resulted in decreased ability of clone formation and cell movement.