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该文旨在探讨肌动蛋白结合蛋白(actin-binding protein,ANLN)对肝癌细胞迁移与侵袭能力的影响。应用荧光定量PCR(q RT-PCR)检测ANLN在肝癌组织和癌旁组织中的表达差异,运用慢病毒介导sh RNA干扰技术靶向敲低肝癌细胞Huh-7中ANLN的表达,并通过q RT-PCR和Western blot方法验证敲低效率;通过细胞迁移实验和侵袭实验检测肝癌细胞的迁移与侵袭能力。进一步通过q RT-PCR和Western blot检测ANLN基因敲低对基质金属蛋白酶9(matrix metallopeptidase 9,MMP9)m RNA和蛋白质水平的影响。最后,分析MMP9在ANLN敲低调控的肝癌细胞迁移侵袭过程中的作用。结果显示,在20例肝癌组织样本和癌旁组织中,ANLN在肝癌组织中m RNA水平较癌旁组织显著增高(P<0.001)。其中,在发生转移的肝癌组织中,ANLN m RNA水平较无转移的肝癌组织显著增高(P<0.001)。慢病毒介导sh RNA能显著抑制肝癌细胞中ANLN的表达,ANLN基因敲低能抑制肝癌细胞的迁移能力,并能显著抑制肝癌细胞的侵袭能力。机制研究发现,ANLN的基因敲低能显著抑制MMP9的表达,MMP9的过表达能逆转ANLN基因敲低对肝癌细胞迁移侵袭能力的抑制作用。该研究结果提示,在肝癌组织中,ANLN m RNA水平明显增高,ANLN的表达水平与迁移侵袭能力密切相关。ANLN基因敲低可能通过调节MMP9的表达,从而抑制肝癌细胞的迁移侵袭能力。
This article aims to investigate the effect of actin-binding protein (ANLN) on the migration and invasion of hepatocellular carcinoma cells. The expression of ANLN was detected by qRT-PCR in hepatocellular carcinoma tissues and adjacent tissues. The expression of ANLN in Huh-7 cells was detected by lentivirus-mediated sh RNA interference. The expression of ANLN was detected by q The knockdown efficiency was verified by RT-PCR and Western blot. The migration and invasion ability of hepatoma cells were detected by cell migration and invasion assays. The effect of ANLN gene knockdown on m RNA and protein levels of matrix metallopeptidase 9 (MMP9) was further analyzed by q RT-PCR and Western blot. Finally, the role of MMP9 in the migration and invasion of hepatocellular carcinoma cells regulated by ANLN knockdown was analyzed. The results showed that m RNA level of ANLN in HCC tissues was significantly higher than that in paracancerous tissues (P <0.001) in 20 specimens of HCC tissues and paracancerous tissues. Among them, the level of ANLN m RNA in metastatic HCC tissues was significantly higher than that in non-metastatic HCC tissues (P <0.001). Lentivirus-mediated sh RNA significantly inhibited the expression of ANLN in hepatocellular carcinoma cells. ANLN knockdown could inhibit the migration of hepatocellular carcinoma cells and significantly inhibit the invasion ability of hepatoma cells. Mechanism study found that ANLN gene knockdown can significantly inhibit the expression of MMP9, MMP9 overexpression can reverse ANLN gene knockdown on the ability of liver cancer cell migration and invasion inhibition. The results suggest that in liver cancer tissue, ANLN m RNA levels were significantly increased, ANLN expression levels and migration and invasion are closely related. ANLN gene knockdown may regulate the expression of MMP9, thereby inhibiting the migration and invasion of liver cancer cells.