膜联蛋白A2调控人脑胶质瘤细胞侵袭迁移的研究

Regulatory effect of annexin A2 on invasion and migration of human glioma cells

目的观察膜联蛋白A2（ANXA2）对人脑胶质瘤细胞侵袭迁移的调控作用。方法将ANXA2小干扰RNA（siRNA）和siRNA control转染到脑胶质瘤细胞U87中记为ANXA2 siRNA组和siRNA-NC组，同时设置Control组，反转录-聚合酶链反应（RT-PCR）检测细胞中ANXA2 mRNA，Western blot检测细胞中ANXA2蛋白表达，噻唑蓝（MTT）法检测细胞增殖，Transwell小室检测细胞迁移和侵袭能力，Western blot检测E-钙黏蛋白（E-cadherin）、基质金属蛋白酶-2（MMP-2）蛋白表达。结果siRNA-NC组细胞中ANXA2 mRNA（0.85±0.08）和蛋白（0.72±0.09）水平与Control组（mRNA和蛋白水平分别为0.87±0.05、0.75±0.07）比较差异无统计学意义（P值分别为0.258与0.751）。siRNA-NC组细胞存活率[（98.87±11.74）%，P＝0.695]、侵袭数目[（40.65±4.85）个，P＝0.410]、迁移数目[（66.87±7.64）个，P＝0.278]及E-cadherin（0.70±0.09，P＝0.320）、MMP-2蛋白（0.29±0.02，P＝0.196）水平与Control组比较差异无统计学意义。ANXA2 siRNA组细胞存活率[（52.98±5.61）%，P＝0.006]、侵袭数目[（23.22±2.25）个，P＝0.002]、迁移数目[（41.68±3.87）个，P＝0.002]、MMP-2蛋白（0.11±0.03，P＝0.001）水平与Control组比较明显降低，E-cadherin蛋白（1.14±0.11，P＝0.002）水平与Control组（0.72±0.06）比较明显升高。结论干扰ANXA2表达能够抑制脑胶质瘤细胞增殖、侵袭和迁移，降低MMP-2表达水平，促进E-cadherin表达。

Objective To investigate the regulatory effect of annexin A2 （ANXA2） on invasion and migration of human glioma cells.Methods The ANXA2 small interfering RNA （siRNA） and siRNA control were transfected into glioma cells U87 and labeled as ANXA2 siRNA group and siRNA-NC group. The control group was set up at the same time. The expression of ANXA2 mRNA and protein was detected by reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting respectively. Cell proliferation was detected by methyl thiazol tetrazolium （MTT）. The cell migration and invasive ability was detected by Transwell. Western blotting was used to detect E-cadherin and matrix metalloproteinase （MMP）-2 proteins.Results The levels of ANXA2 mRNA （0.85±0.08, P=0.258） and protein （0.72±0.09, P=0.751） in siRNA-NC group were not statistically differed from those in the control group （for mRNA and protein： 0.87±0.05 and 0.75±0.07）. The survival rate [（98.87±11.74）%, P=0.695], the number of invasive cells （40.65±4.85, P=0.410）, the number of migrating cells （66.87±7.64, P=0.278） and the levels of E-cadherin （0.70±0.09, P=0.320） and MMP-2 protein （0.29±0.02, P=0.196） in siRNA-NC group showed no statistically significant difference from those in the control group. The survival rate [（52.98±5.61）%, P=0.006], the number of invasive cells （23.22±2.25, P=0.002）, the number of migrating cells （41.68±3.87, P=0.002） and the levels of MMP-2 protein （0.11±0.03, P=0.001） in the ANXA2 siRNA group were significantly lower, and the levels of E-cadherin protein （1.14±0.11, P=0.002） were significantly higher than those in the control group （0.72±0.06）.Conclusion Interference with ANXA2 can inhibit the proliferation, invasion and migration of glioma cells, reduce the expression level of MMP-2 and promote the expression of E-cadherin.