HOXA9基因抑制肝癌细胞HepG2迁移与侵袭能力的实验研究

摘要

Objective:To investigate the inhibitory effects of HOXA9 gene on migration and invasion of hepatocel-lular carcinoma cell line HepG2. Methods:Eukaryotic vector expressing HOXA9 gene and empty vector were trans-fected into hepatocellular carcinoma cell line HepG2 by EndoFectinTM -Max transfection reagents. Liposome mediated method was used to transfect siRNA sequence targeting HOXA9 and control sequence into HepG2 cells. HOXA9 ex-pression levels after transfection were detected by using Real-time PCR and Western blot methods. The tumor cell a-bilities of migration and invasion after transfection were measured by wound scratch and transwell assays. Results:The mRNA and protein levels of HOXA9 were significantly increased after transfection of Eukaryotic vector expressing HOXA9 gene. Wound scratch assay revealed the ability of cell mobility was significantly reduced after transfecting of HOXA9 gene,and Transwell assay showed the number of migratory cells in HOXA9 over-expressing group was sig-nificantly reduced. While after transfecting siRNA sequence targeting HOXA9,the mRNA and protein levels of HOXA9 were significantly reduced. Wound scratch assay revealed the ability of cell mobility in siRNA group was sig-nificantly increased,and Transwell assay showed the number of migratory cells in siRNA group was significantly in-creased. Conclusion:HOXA9 gene inhibit migration and invasion abilities of HepG2 cell,which can provide an exper-iment basis for investigating effects of HOXA9 in the occurrence and development of hepatocellular carcinoma.%目的：探讨同源盒基因HOXA9对肝细胞肝癌细胞株HepG2迁移与侵袭能力的影响。方法：采用EndoFectinTM -Max转染试剂将HOXA9基因真核表达载体及其空载体转染至HepG2细胞，并应用脂质体法转染HOXA9 siRNA序列及阴性对照序列，采用实时荧光定量PCR（ Real-time PCR）及Western blot方法检测转染后HOXA9的mRNA和蛋白表达水平。以划痕实验和Transwell方法检测转染后细胞的迁移和侵袭能力。结果：在肝癌细胞HepG2中瞬时转染HOXA9真核表达载体后，其mRNA和蛋白表达明显增加，划痕实验检测发现48h细胞愈合率明显抑制，Transwell实验发现迁移和侵袭至下室的细胞数目明显减少；而转染HOXA9 siRNA序列后，HepG2细胞中HOXA9 mRNA及蛋白表达明显下调，划痕实验显示48h划痕愈合率增加，Tran-swell实验显示迁移和侵袭至下室的细胞数目明显增多。结论：HOXA9基因对肝癌细胞HepG2的迁移和侵袭能力有明显抑制作用，为进一步研究HOXA9在肝细胞肝癌发生和进展中的作用奠定了基础。