膜联蛋白A7对肿瘤细胞黏附分子表达及生物学行为的影响

Effect of Annexin A7 on Expression of Adhesion Molecules and Biological Behavior in Mouse Hepatocarcinoma Hca-P Cells

[目的]研究Annexin A7基因对肿瘤细胞黏附分子表达及其生物学行为的影响。[方法]采用sh RNA干扰技术稳定转染小鼠Hca-P细胞,获得Annexin A7下调表达及Hca-P无关序列细胞株,采用Western Blot、q RT-PCR分别检测Annexin A7对黏附相关分子FAK、Src、E-cadherin等基因及蛋白表达水平的影响,应用淋巴结粘附及Transwell侵袭实验检测Annexin A7下调对Hca-P细胞淋巴结粘附及侵袭能力的干扰。[结果]Annexin A7基因表达下调组细胞的FAK和Src基因及蛋白表达水平较A7无关序列组分别上升1.81倍、1.80倍及1.58倍、1.59倍（P〈0.05）,E-cadherin基因及蛋白表达水平则分别下降54.11%和54.27%,而上述分子基因与蛋白表达水平在未处理Hca-P组与A7无关序列组间无统计学差异（P〉0.05）,且在Annexin A7下调后Hca-P细胞的淋巴结粘附及侵袭能力上升,Sh RNA-Annexin A7组淋巴结黏附细胞数（1074±162.8）显著性多于A7无关序列组（234.7±40.08）及Hca-P组（220.3±39.53）（P〈0.05）,Sh RNA-A7细胞组穿过小室的细胞数目（294.8±64.40）明显多于A7无关序列细胞组（79±28.04）及Hca-P细胞组（57.60±25.62）（P〈0.05）,而Hca-P与无关序列细胞组之间则无统计学差异（P〉0.05）。[结论]Annexin A7可通过影响黏附相关分子FAK、Src、E-cadherin等基因与蛋白表达水平从而改变Hca-P细胞生物学行为。

[Objective] To investigate the effect of Annexin A7 on the expression of adhesion molecules and biological behavior in mouse hepatocarcinoma cells. [Methods] Mouse heparocarcinoma Hca-P cells were transfected with Annexin A7 sh RNA（Annexin A7-sh RNA group） or unrelated sh RNA（blank control group）. The expressions of Annexin A7 and the adhesion molecules FAK,Src,E-cadherin in Hca-P cells were detected by Western Blot and q RT PCR. Transwell assay and lymph node adhesion testing were used to analyze the invasion and adhesion ability of Hca-P cells after Annexin A7 sh RNA interference. [Results] Compared with the blank control group,the expression of Annexin A7 was reduced in Annexin A7-sh RNA group（P〈0.05）; the m RNA and protein expression levels of FAK and Src were increased by 1.81,1.80 times and1.58,1.59 times（P〈0.05）,respectively,while the m RNA and protein expression levels of E-cadherin were decreased by 54.11% and 54.27%,respectively（P〈0.05）. Meanwhile,the invasion and adhesion ability in Annexin A7-sh RNA group was significantly higher than wild Hca–P cells and blank control group,the number of lymph node cells in Sh RNA-Annexin A7 group（1074 ± 162.8）was significantly higher than that in blank control group（234.7 ± 40.08） and wild Hca-P cell group（220.3±39.53）（P〈0.05）,the number of migrant cells in Sh RNA-Annexin A7 group（294.8±64.40）was significantly higher than that in blank control group（79 ±28.04） and wild Hca-P cell group（57.60±25.62）（P〈0.05）,while there was no significant difference between wild Hca-P cell and blank control groups（P〈0.05）. [Conclusion] Annexin A7 may change biological behavior of hepatocarcinoma Hca-P cells by influencing the gene and protein expression of adhesion molecules FAK,Src and E-cadherin.