天津医药 ›› 2019, Vol. 47 ›› Issue (5): 468-472.doi: 10.11958/20182189

• 细胞与分子生物学 • 上一篇    下一篇

靶向调控AQP4对非小细胞肺癌细胞化疗敏感性的调控作用

郭守俊 1△,谢传华 1,黄萍 1,邱伊连 1,王硕 1,班振英 2,张威 2   

  1. 基金项目:河南省高等学校重点科研项目(17A310027) 作者单位:1赣州市肿瘤医院内一科(邮编341000);2郑州大学第三附属医院病理科 作者简介:郭守俊(1971),男,学士,主任医师,主要从事胸部肿瘤的临床诊断、治疗研究 △通讯作者 E-mail:jxgzguosj@126.com
  • 收稿日期:2019-01-02 修回日期:2019-04-01 出版日期:2019-05-15 发布日期:2019-05-15
  • 通讯作者: 郭守俊 E-mail:315617735@qq.com
  • 基金资助:
    河南省高等学校重点科研项目

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

GUO Shou-jun1△, XIE Chuan-hua1, HUANG Ping1, QIU Yi-lian1, WANG Shuo1, BAN Zhen-ying2, ZHANG Wei2   

  1. 1 Department of Medical Oncology, Ganzhou Cancer Hospital, Ganzhou 341000, China; 2 Department of Pathology, the Third Affiliated Hospital of Zhengzhou University △Corresponding Author E-mail: jxgzguosj@126.com
  • Received:2019-01-02 Revised:2019-04-01 Published:2019-05-15 Online:2019-05-15

摘要: 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。

关键词: 非小细胞肺癌, 靶向调控, 耐药性, AQP4, RNA干扰

Abstract: ponding Author E-mail: jxgzguosj Abstract: Objective To investigate the effect of targeting silenced aquaporin 4 (AQP4) gene on the chemotherapy sensitivity of non-small cell lung cancer (NSDCLC) and its mechanism. Methods The qRT-PCR method and Western bolt assay were used to detect the expression of AQP4 in A549 and A549/DDP cell lines (human cisplatin-resistant NSDCLC cell line). The siRNA-1 and siRNA-2 targeting AQP4 were designed. AQP4 expression was blocked by the siRNA in A549/DDP cells. The optimal siRNA was screened and transfected into A549/DDP cells. siRNA-NC and blank cells were set up as controls. The proliferation and half inhibitory concentration (IC50) of A549/DDP cells were detected by CCK8 method. The apoptosis 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 increased in A549 /DDP cells compared with A549 cells. After silencing AQP4 expression, the proliferation was inhibited and the apoptosis rate was significantly increased in A549 / DDP cells. Silencing AQP4 expression significantly increased the sensitivity to cisplatin in lung cancer cells. AQP4 knockdown could also up-regulate the expression of P53, and down regulate the expression of Bcl-2. Conclusion The selective targeting of the AQP4 expression can increase the sensitivity of NSDCLC cells to chemotherapeutic drugs.

Key words: NSCLC, target regulating, durg-resistance, AQP4, RNA interference