孕激素和脂联素分子受体11对A549肺癌细胞侵袭转移的影响研究

Study of the effect of PAQR11 on invasion and metastasis in A549 lung cancer cells

目的:观察孕激素和脂联素分子受体11(PAQR11)对A549肺癌细胞侵袭转移的作用,探索其可能的分子机制。方法:设计2条干扰PAQR11的核酸序列,以分子克隆的方法插入p LKO.1慢病毒载体,转染HEK-293T细胞包装病毒,以病毒上清感染A549细胞,嘌呤霉素筛选A549细胞;细胞划痕实验检测敲低PAQR11对A549细胞迁移的影响;Transwell小室实验检测敲低PAQR11对A549细胞侵袭能力的影响;实时定量PCR检测上皮间质转化相关分子的表达。结果:成功构建了干扰PAQR11表达慢病毒载体,获得稳定干扰PAQR11的A549细胞,实时定量PCR、Western blot验证干扰效果。敲低PAQR11能够促进A549细胞的迁移和侵袭能力;敲低PAQR11后E-Cadherin的表达水平降低,Snail1、Twist1和N-Cadherin表达升高。结论:敲低PAQR11能够促进A549细胞的迁移和侵袭能力,其机制可能与EMT增强有关。

Objective： To investigate the effect of PAQR11 on invasion and metastasis in A549 lung cancer cells, and explore the possible molecular mechanism. Methods： Two oligonucleotides for interfering PAQR11 were designed, and were cloned into p LKO.1 lentiviral plasmid. entiviral particles were produced with HEK-293 T cells. A549 cells were infected with lentiviral particle solution supernatant and screened with puromycin. Then the migration of PAQR11 knockdown A549 cell was detected with erasion trace test and the invasion was observed with transwell chamber. The expression of EMT associated molecular was detected by real-time PCR. Results： Lentiviral vector for interfering PAQR11 was built. Stable PAQR11 knockdown A549 lung cancer cells were established. The knockdown of PAQR11 was tested by real-time PCR and Western blot. Cell migration and invasion was promoted in PAQR11 knockdown A549 lung cancer cells. PAQR11 knockdown could reduce the expression of E-Cadherin, but up-regulate the level of Snail1, Twist1 and N-Cadherin. Conclusion： Knockdown of PAQR11 could promote the invasion and metastasis of A549 lung cancer cells, and this effect may be relevant with reinforcing EMT.