shRNA沉默AnnxinA2基因对放射抗拒鼻咽癌细胞迁移和侵袭的影响

Effects of sh RNA-mediated silencing of Annexin A2 on migration and invasion of radioresistant nasopharyngeal carcinoma cells

目的探讨sh RNA沉默Annxin A2基因对放射抗拒鼻咽癌细胞迁移和侵袭的影响。方法采用Fu GENE HD将Annexin A2 sh RNA转染放射抗拒的鼻咽癌CNE2(R743)细胞,q RT-PCR验证转染细胞中Annxin A2基因的表达,细胞划痕实验和Transwell实验观察下调Annxin A2基因表达及联合照射对鼻咽癌细胞迁移和侵袭的影响,Western blot检测细胞中基质金属蛋白酶2(MMP2)蛋白的表达。结果转染组细胞Annexin A2基因的m RNA相对表达量为(0.25±0.17),较对照组的(1±0.00)和转染对照组的(0.96±0.06)明显下降,差异有统计学意义(P<0.05)。细胞划痕实验显示,下调Annxin A2基因表达可抑制照射诱导的CNE2(R743)细胞的迁移能力(P<0.05)。Transwell实验结果显示,下调Annxin A2基因表达可抑制照射诱导的CNE2(R743)细胞的迁移和侵袭能力(P<0.05)。Western blot结果表明,下调Annxin A2基因表达能抑制照射诱导的MMP2蛋白的表达上调(P<0.05)。结论下调Annxin A2基因表达能抑制照射诱导放射抗拒的鼻咽癌CNE2(R743)细胞的迁移和侵袭能力,可能与MMP2蛋白表达下调相关。

Objective To investigate the effect of shRNA-mediated silencing of Annexin A2 and combined irradiation on the migration and invasion of mdioresistant nasopharyngeal carcinoma cells. Methods Annexin A2 shRNA was transfected into CNE2（R743） cells using FuGENE HD. The expression of Annexin A2 gene in transfected cells was verified by qRT-PCR. The effect of down-regulation of Annexin A2 expression cornhined with irradiation on the migration and invasion of nasopharyngeal carcinoma cells were determined by scratch assay and transwell chamber assay. The expression of MMP2 was detected by Western blot. Results The relative expression of Annexin A2 mRNA in transfected group（0.25±0.17 ） was significantly lower than in control group（ 1±0.00） and transfected control group（0.96±0.06）, the difference was statistically significant （P〈0.05）. Cell scratches showed that down-regulation of Annexin A2 could inhibit the migration of CNE2（ R743 ） cells induced by irmdiation（P〈0.05）. Transwell assay showed that down-regulation of Annexin A2 expression inhibited the migration and invasion of CNE2 （ R743 ） cells induced by irradiation （P〈0.05）. Western blot analysis showed that down-regulation of Annexin A2 could inhibit the up-regulation of MMP2 protein expression induced by irradiation （P〈0.05）. Conclusion Down-regulation of Annexin A2 can inhibit the migration and invasion of CNE2（ R743 ） ceils induced by irradiation, which may be related to the down-regulation of MMP2 protein expression.