中文摘要：

目的观察下调膜联蛋白A2（annxin A2，ANXA2）表达对肝癌（HCC）细胞生物学行为的影响。方法以定量PCR分析不同肝癌细胞ANXA2 mRNA转录水平，以Western blot分析ANXA2表达；以免疫荧光检测ANXA2在细胞内表达和分布；以碘化丙啶染色和流式细胞术分析细胞周期；以CCK-8试剂盒分析细胞增殖潜能；以transwell实验分析其侵袭潜能；以创伤愈合试验分析其迁移潜能；以裸鼠移植瘤模型分析其致瘤潜能。两样本均数比较用t检验，率的比较用x^2检验，等级资料比较用秩和检验，多样本均数的两两比较采用口检验，组间差异用单因素方差分析。结果高侵袭潜能MHCC97-H细胞ANXA2表达明显高于HepG2、SMMC-7721和SMMC-7402和L02细胞；特异性shRNA沉默ANXA2效率在80％以上；免疫荧光显示ANXA2定位于细胞膜和细胞质，胞核少见；下调ANXA2表达明显下调HCC细胞s期比例（口=8．001，P=0．002），抑制细胞增殖（q=17．140，P＜0．01）、迁移潜能（q=12．808，P＜0．01）和侵袭潜能（q=9．069，P=0．002）；裸鼠移植瘤模型显示干扰组瘤重下降（g=11．968，P＜0．01），瘤体中ANXA2表达下调（Z=2．530，P=0．011）。结论下调ANxA2基因转录显著影响肝癌细胞的生物学行为，可望成为肝癌分子治疗的潜在靶目标。

英文摘要：

Objective To investigate the effects of Annexin A2 （ANXA2） deficiency on the malignant biological behaviour of hepatoma cells. Methods The human hepatocellular carcinoma （HCC） cells lines MHCC97-H, HepG2, SMMC-7721, SMMC-7402 and L02 were evaluated. The expression and distribution of ANXA2 were analysed by westem blotting, real-time PCR, immunofluorescence and immunohistochemislry. Cell cycle was assessed by flow cytometry and propidium iodide staining. Effects of ANXA2 silencing on invasion and migration potential were assessed by transwell assay and wound healing assay, respectively. Proliferative potential was assessed by CCK-8 kit in vitro and xenograft tumour-growth assay in vivo. The t -test, chi square test, rank sum test, q-test and F-test were used for statistical analyses. Results The expression level of ANXA2 was markedly higher in the MHCC97-H cells with high metastasis potential than in the HepG2, SMMC-7721, SMMC-7402 and L02 cells. The efficiency of shRNA-mediated ANXA2 deficiency was more than 80% analysis of the MHCC97-H cells indicated that ANXA2 expression was mainly localized to the cellular membrane and cytoplasm, with some nuclear localization. Down-regulation of ANXA2 led to S-phase arrest of HCC cells （q = 8.001, P = 0.002） and an inhibition of proliferation （q = 17.140, P 〈0.01）, migration （q = 12.808, P 〈0.01） and invasion potential （q = 9.069, P = 0.002）. Xenograft tumour-growth assay indicated that shRNA targeting of ANXA2 led to lower tumour weight （q = 11.968, P 〈0.001） and down-regulated ANXA2 expression （Z = 2.530,P = 0.011）. Conclusion Down-regulation of Annexin A2 gene transcription effectively changes the biological behaviottrs ofhepatoma cells, and may represent a potential target of HCC molecular therapies.