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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Endo FectinTM-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HOXA9siRNA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,Transwell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。
Objective: To investigate the effect of homeobox gene HOXA9 on the migration and invasion of hepatocellular carcinoma cell line HepG2. Methods: The HOXA9 gene eukaryotic expression vector and its empty vector were transfected into HepG2 cells by Endo FectinTM-Max transfection reagent, and HOXA9 siRNA sequence and negative control sequence were transfected by lipofectamine. Real-time fluorescence quantitative PCR -time PCR) and Western blot were used to detect the mRNA and protein expression of HOXA9 after transfection. Scratch assay and Transwell method were used to detect the migration and invasion ability of transfected cells. Results: The mRNA and protein expression of HOXA9 eukaryotic expression vector was transiently transfected into HepG2 hepatoma cells. The scratch healing assay showed that the cell healing rate was obviously inhibited at 48h. Transwell assay showed that the number of cells migrated and infiltrated into the lower chamber was significantly While the expression of HOXA9 mRNA and protein in HepG2 cells was significantly down-regulated after transfection of HOXA9 siRNA sequence. Scratch experiments showed that the wound healing rate increased after 48 h, and the number of cells migrated and infiltrated into the lower compartment was significantly increased by Transwell assay. CONCLUSION: The HOXA9 gene has a significant inhibitory effect on the migration and invasion of HepG2 cells, which lays the foundation for the further study on the role of HOXA9 in the occurrence and progression of hepatocellular carcinoma.