Department of Medical Ultrastructure, Xiangya School of Medicine, Central South University, Changsha, Hunan, 410078, People's Republic of China.
Pathol Oncol Res. 2012 Apr;18(2):183-90. doi: 10.1007/s12253-011-9427-2. Epub 2011 Jun 18.
In the previous study, we found that Annexin A2 was significantly up-regulated in lung cancer and could induce related-antigen in lung cancer patients' serum. To further study the function of Annexin A2, the short hairpin RNA plasmid targeting Annexin A2 was constructed in vitro and transfected into human lung adencarcinoma A549 cells. Knocking down Annexin A2 expression by shRNA, the mRNA level of Annexin A2 was investigated by semi-quantitative RT-PCR. The expression of Annexin A2 protein was examined by Western Blotting and Immuocytochemistry. MTT assay and Transwell chamber model were used to evaluate proliferation and invasion of A549 cells in vitro. The concentration of matrix metalloproteinase-2 (MMP-2) and cathepsin B (CB) in the supernatant was evaluated by ELISA. At 48 h after transfection, the expression of Annexin A2 mRNA and protein was down-regulated significantly, respectively (p < 0.05).The proliferation and invasion capability of A549 cells also decreased significantly (p < 0.05). The concentration of MMP-2 and CB was down-regulated obviously, respectively (p < 0.05). This study implies that Annexin A2 might play an important role in the progression and invasion of human lung cancer cells, and could promote progression of lung cancer by regulating the expression of MMP-2 and CB.
在之前的研究中，我们发现 Annexin A2 在肺癌中显著上调，并能诱导肺癌患者血清中的相关抗原。为了进一步研究 Annexin A2 的功能，我们在体外构建了靶向 Annexin A2 的短发夹 RNA 质粒，并转染入人肺腺癌细胞 A549 中。通过 shRNA 敲低 Annexin A2 的表达，用半定量 RT-PCR 检测 Annexin A2 的 mRNA 水平。用 Western Blotting 和免疫细胞化学检测 Annexin A2 蛋白的表达。用 MTT assay 和 Transwell 室模型评估 A549 细胞在体外的增殖和侵袭能力。用 ELISA 评估上清液中基质金属蛋白酶-2 (MMP-2) 和组织蛋白酶 B (CB) 的浓度。转染 48 小时后， Annexin A2 的 mRNA 和蛋白表达均显著下调(p < 0.05)。A549 细胞的增殖和侵袭能力也显著下降(p < 0.05)。MMP-2 和 CB 的浓度明显下调(p < 0.05)。本研究表明 Annexin A2 可能在人类肺癌细胞的进展和侵袭中发挥重要作用，并通过调节 MMP-2 和 CB 的表达促进肺癌的进展。
