Zhai Yang, Chen Qian, Wang Yuzhen, Li Xu, Li Lina
Department of Oncology, Tumor hospital of Shaanxi Province, Xi'an 710061, China.
Department of Reproductive Medicine, The First Affiliated Hospital of Xian Jiaotong University, Xi'an 710061, China.
Zhongguo Fei Ai Za Zhi. 2020 Sep 20;23(9):762-771. doi: 10.3779/j.issn.1009-3419.2020.103.13. Epub 2020 Aug 10.
Lung cancer is the most common cancer worldwide with the highest morbidity and mortality, in which the non-small cell lung cancer accounts for 80% of all cases. The expression of (HOX transcript antisense RNA) HOTAIR were abnormal in a variety of tumor tissues and is involved in the regulation of the occurrence and development of lung cancer. The purpose of this study is to investigate the effect and mechanism of down-regulation of HOTAIR on gefitinib resistance of lung adenocarcinoma HCC827 cells by targeting PTEN.
The HOTAIR downstream target gene was predicted by bioinformatics database. The small interfering RNAs (siRNA) which is corresponding to HOTAIR was transfected using Lipofectamine™ 2000. Quantitative real-time PCR (RT-qPCR) and Western blot were used to detect the expression of HOTAIR, PTEN, PI3K and AKT in HCC827 and HCC827GR cells. MTT assay was used to detect the changes in drug resistance of HCC827GR cells. Flow cytometry analysis were used to test the cell proliferation and the rate of apoptosis.
The expression of HOTAIR increased in HCC827GR and the serum of NSCLC patients with gefitinib resistance (P<0.05). Transfection of HOTAIR siRNA decreased the expression of HOTAIR (P<0.05), and increased the expressions of PTEN (P<0.05), while the expression of PI3K and AKT were decreased (P<0.05). Compared with the blank control group, down-regulation of HOTAIR increased the sensitivity of HCC827GR cells to gefitinib. The cell proliferation ability was decreased and the apoptosis was promoted apparently (P<0.05).
Down-regulation of HOTAIR can suppress the cell growth and promote the apoptosis, and it can reverse the resistance of HCC827GR cells to gefitinib. Its potential mechanism may be related with the targeting of PTEN/PI3K/AKT pathway.
肺癌是全球最常见的癌症，发病率和死亡率最高，其中非小细胞肺癌占所有病例的80%。（HOX转录本反义RNA）HOTAIR在多种肿瘤组织中表达异常，并参与肺癌发生发展的调控。本研究旨在探讨下调HOTAIR通过靶向PTEN对肺腺癌HCC827细胞吉非替尼耐药性的影响及机制。
通过生物信息学数据库预测HOTAIR下游靶基因。使用Lipofectamine™ 2000转染与HOTAIR对应的小干扰RNA（siRNA）。采用定量实时聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测HCC827和HCC827GR细胞中HOTAIR、PTEN、PI3K和AKT的表达。采用MTT法检测HCC827GR细胞耐药性的变化。采用流式细胞术分析检测细胞增殖和凋亡率。
HOTAIR在HCC827GR细胞及吉非替尼耐药的非小细胞肺癌患者血清中的表达升高（P<0.05）。转染HOTAIR siRNA后HOTAIR表达降低（P<0.05），PTEN表达升高（P<0.05），而PI3K和AKT表达降低（P<0.05）。与空白对照组相比，下调HOTAIR可增加HCC827GR细胞对吉非替尼的敏感性。细胞增殖能力降低，凋亡明显增加（P<0.05）。
下调HOTAIR可抑制细胞生长并促进凋亡，可逆转HCC827GR细胞对吉非替尼的耐药性。其潜在机制可能与靶向PTEN/PI3K/AKT通路有关。
