靶向调控AQP4对非小细胞肺癌细胞化疗敏感性的调控作用

摘要：目的 探讨靶向沉默水通道蛋白4（AQP4）基因对非小细胞肺癌细胞化疗敏感性影响，并研究其分子作用机制。方法 实时荧光定量 PCR（qRT-PCR）和 Western blot 分别检测非小细胞肺癌 A549 细胞以及顺铂耐药菌株A549/DDP中AQP4 mRNA和蛋白的表达。设计靶向AQP4的siRNA-1和siRNA-2，采用siRNA抑制A549/DDP细胞中AQP4的表达，筛选出最优siRNA，将其转染A549/DDP细胞，设立siRNA-NC组和空白对照组。CCK-8法检测细胞增殖能力并计算各组细胞的半数抑制浓度（IC50）；流式细胞术检测细胞凋亡。Western blot检测P53和Bcl-2蛋白的表达。结果 AQP4 mRNA和蛋白在A549/DDP细胞中的表达水平显著高于A549细胞，AQP4表达沉默后A549/DDP细胞增殖抑制，细胞凋亡增加，对顺铂的敏感性增加，凋亡相关蛋白P53表达增加，而Bcl-2蛋白表达降低。结论 通过靶向调控AQP4的表达可以增加非小细胞肺癌细胞对化疗药物的敏感性。

The effect of silencing AQP4 expression on the chemotherapy sensitivity ofnon-small cell lung cancer

ponding Author E-mail: jxgzguosjAbstract: Objective To investigate the effect of targeting silenced aquaporin 4 (AQP4) gene on the chemotherapysensitivity of non-small cell lung cancer (NSDCLC) and its mechanism. Methods The qRT-PCR method and Western boltassay were used to detect the expression of AQP4 in A549 and A549/DDP cell lines (human cisplatin-resistant NSDCLC cellline). The siRNA-1 and siRNA-2 targeting AQP4 were designed. AQP4 expression was blocked by the siRNA in A549/DDPcells. The optimal siRNA was screened and transfected into A549/DDP cells. siRNA-NC and blank cells were set up ascontrols. The proliferation and half inhibitory concentration (IC50) of A549/DDP cells were detected by CCK8 method. Theapoptosis of U251 cells was detected by AnnexinV- FITC/PI double marker flow cytometry. The expressions of P53 and Bcl-2 were detected by Western blot assay. Results The expressions of AQP4 mRNA and protein were significantly increasedin A549 /DDP cells compared with A549 cells. After silencing AQP4 expression, the proliferation was inhibited and theapoptosis rate was significantly increased in A549 / DDP cells. Silencing AQP4 expression significantly increased thesensitivity to cisplatin in lung cancer cells. AQP4 knockdown could also up-regulate the expression of P53, and downregulate the expression of Bcl-2. Conclusion The selective targeting of the AQP4 expression can increase the sensitivity ofNSDCLC cells to chemotherapeutic drugs.