【摘要】 目的：研究低氧条件对Hep3B肝癌细胞增殖、迁移的影响，探讨低氧对上皮—间质转化（EMT）、糖异生及脂肪酸合成相关基因的表达变化及意义。方法：克隆形成实验研究Hep3B的增殖能力，细胞划痕愈合和Transwell迁移实验研究Hep3B的迁移能力。肿瘤基因组图谱数据库（TCGA）和基因表达综合数据库（GEO）分析EMT相关基因SNAl1、SNAl2、CDH1、CDH2、ZEB1和Vimentin，糖异生相关基因PCK2、G6PC及脂肪酸合成相关基因ACACA的mRNA在肝细胞癌（HCC）与癌旁组织中的表达情况，并使用GEO数据库分析上述基因在低氧条件下的mRNA表达。实时荧光定量聚合酶链式反应（RT-qPCR）检测上述基因在常氧和低氧条件下培养的Hep3B细胞中的mRNA表达情况。结果：低氧环境可以促进Hep3B细胞的增殖和迁移能力。与癌旁组织比较，TCGA和GEO数据库均显示，ZEB1和ACACA在肝癌组织中表达上调，G6PC和PCK2则下调，SNAl2和Vimentin仅在TCGA中显示上调，SNAl1和CDH1仅在GEO显示下调。RT-qPCR结果显示，与常氧组相比，低氧环境下的Hep3B细胞中EMT相关基因SNAl1、CDH1、CDH2的表达量降低（P<0.05），而SNAl2、Vimentin的表达量则升高（P<0.05），糖异生相关基因PCK2、G6PC和脂肪酸合成相关基因ACACA在低氧环境中的表达量均降低（P<0.05），而ZEB1的表达量差异无统计学意义（P>0.05）。结论：低氧微环境可以调节肝癌Hep3B细胞中EMT相关基因SNAl1、SNAl2、CDH1、CDH2和Vimentin以及代谢相关基因ACACA、G6PC和PCK2的表达，进而促进Hep3B细胞的增殖和迁移。更多还原

【Abstract】 Objective: To study the effects of hypoxic condition on the proliferation and migration of Hep3B hepatocellular carcinoma cells, and to explore the effects of hypoxia on the expression of genes related to epithelialmesenchymal transition(EMT), gluconeogenesis and fatty acid synthesis as well as their significance. Methods:Clonogenic assay was performed to study the proliferation ability of Hep3B, and cell scratch and Transwell migration assays were performed to study the migration ability of Hep3B. The mRNAs of EMT-re-lated genes SNAl1, SNAl2, CDH1, CDH2, ZEB1 and Vimentin, gluconeogenesis-related genes PCK2, G6PC and fatty acid synthesis-related gene ACACA were analyzed in hepatocellular carcinoma(HCC) and normal tissues adjacent to the tumor(NAT) using the Cancer Genome Atlas(TCGA) and Gene Expression Omnibus(GEO) databases. The mRNA expression of these genes under hypoxic condition was analyzed using the GEO database. Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR) was performed to detect the mRNA expression of the above genes in Hep3B cells cultured under normoxic and hypoxic conditions. Results: The hypoxic condition promoted the proliferation and migration ability of Hep3B cells. Compared with the NAT, both TCGA and GEO databases showed that ZEB1 and ACACA expression was upregulated in HCC tissues, while G6PC and PCK2 were downregulated; SNAl2 and Vimentin showed upregulation only in TCGA, while SNAl1 and CDH1 downregulation only in GEO. RT-qPCR results showed that the expression of EMT-related genes SNAl1,CDH1 and CDH2 decreased(P<0.05), while the expression of SNAl2 and Vimentin increased(P<0.05) in Hep3B cells under hypoxic condition compared with the normoxic group; the expression of gluconeogenesis-related genes PCK2, G6PC and fatty acid synthesis-related gene ACACA was reduced under the hypoxic condition(P<0.05); the difference in the expression of ZEB1 was not statistically significant(P>0.05). Conclusion: Hypoxic microenvironment can regulate the expression of EMT-related genes SNAl1, SNAl2, CDH1, CDH2 and Vimentin and metabolism-related genes ACACA, G6PC and PCK2 in hepatocellular carcinoma Hep3B cells, which in turn promotes the proliferation and migration of Hep3B cells.更多还原