小分子NEK2干扰RNA对肺癌细胞耐药的研究.docVIP

小分子NEK2干扰RNA对肺癌细胞耐药的研究 　　[摘要] 目的 探讨小分子NEK2干扰RNA（siRNA-NEK2）逆转人肺癌耐药细胞（A549/DDP）耐药性的研究。 方法 应用Real-time PCR法及Western blot法验证NEK2基因在人肺腺癌耐药细胞系（A549/DDP）细胞系呈现高表达；并采用脂质体为转染介质，将NEK2小片段RNA导入A549/DDP细胞沉默NEK2基因；应用MTT法观察顺铂、阿霉素对沉默NEK2基因后的A549/DDP细胞的细胞毒活性变化。 结果 NEK2-mRNA、NEK2蛋白在A549/DDP细胞系表达水平均显著高于A549细胞系，差异有统计学意义（P 0.05）；顺铂、阿霉素对A549/DDP细胞的半数抑制浓度（IC50）分别是（53.08±0.56）、（8.09±0.31）μg/mL，对siRNA-control A549/DDP细胞的IC50分别是（53.09±0.78）、（8.10±0.98）μg/mL，而对siRNA-NEK2 A549/DDP细胞的IC50有显著下降，其值分别是（18.59±0.10）、（2.30±0.14）μg/mL，两组比较，差异均有统计学意义（P 0.05）。 结论 小分子NEK2干扰RNA沉默NEK2可以提高耐药的肺癌细胞对化疗药物的敏感性。NEK2可能成为逆转肺癌耐药的一个新的靶点。 　　[关键词] 肺癌；细胞耐药；NEK2；siRNA-NEK2 　　[中图分类号] R563.9 [文献标识码] A [文章编号] 1673-7210（2014）02（b）-0017-04 　　Study of drug-resistant effect on lung cancer cells with siRNA-NEK2 silencing 　　CHEN Yongfeng1，2 WANG Haijing1 LIU Dingyuan1 ZHOU Xiangdong1 　　1.Department of Respiratory， Southwest Hospital of the Third Military Medical University， Chongqing 400038， China； 2.Department of Reception， the 75600 Army Hospital of PLA， Guangdong Province， Shenzhen 518048， China 　　[Abstract] Objective To investigate the drug-resistant effect of siRNA-NEK2 on lung cancer cells lines A549/DDP. Methods Expression of NEK2-mRNA and NEK2-protein was examined in human A549 cells and A549/DDP cells by Real-time PCR and Western blot. The changes of cytotoxic activity were observed on the A549/DDP cells of silence NEK2 by MTT assay. Results The expression of NEK2-mRNA and NEK2 protein were higher in A549/DDP cells than A549 cells （P 0.05）. After treatment with DDP and Epirubicin， the IC50 values of A549/DDP cells were （53.08±0.56） and （8.09±0.31） μg/mL respectively， and the IC50 values of siRNA-control A549/DDP cells were （53.09±0.78） and （8.10±0.98） μg/mL respectively. While the IC50 values were significantly decreased in siRNA-NEK2 A549/DDP cells [（18.59±0.10）， （2.30±0.14） μg/mL]， two groups were compared， the differences were statistically significant （P 0.05）. Conclusion Silence NEK2 with interfering