实用医学杂志 ›› 2023, Vol. 39 ›› Issue (5): 557-563.doi: 10.3969/j.issn.1006⁃5725.2023.05.006

• 专题报道 • 上一篇    下一篇

LncRNA SNHG15通过miR⁃483⁃3p/DDIT4轴调节非小细胞肺癌细胞的顺铂敏感性和上皮间质转化 

王韵 宋姗 彭斐 胡文霞    

  1. 河北医科大学第四医院 呼吸内科(石家庄 050011)

  • 出版日期:2023-03-10 发布日期:2023-03-10
  • 通讯作者: 胡文霞 E⁃mail:sm_art@sohu.com
  • 基金资助:
    河北省医学科学研究课题计划(编号:20221317)

LncRNA SNHG15 regulates cisplatin sensitivity and epithelial ⁃ mesenchymal transition in non ⁃ small cell lung cancer cells through miR⁃483⁃3p/DDIT4 axis

WANG Yun,SONG Shan,PENG Fei,HU Wenxia.   

  1. Depart⁃ ment of Respiratory Medicine,the Fourth Hospital,Hebei Medical University,Shijiazhuang 050011,China

  • Online:2023-03-10 Published:2023-03-10
  • Contact: HU Wenxia E⁃mail:sm_art@sohu.com

摘要:

目的 探究 LncRNA SNHG15 对非小细胞肺癌(NSCLC)细胞的顺铂(DDP)敏感性和上皮间 质转化的影响以及 miR⁃483⁃3p/DNA 损伤诱导转录物 4(DDIT4)轴在其中发挥的作用方法 体外培养 NSCLC 细胞(A549)、NSCLC 耐药细胞(A549/DDP),检测两种细胞对 DDP IC50 LncRNA SNHG15 miR⁃483⁃3p、DDIT4 mRNA 与蛋白表达,将 A549/DDP 细胞随机分为 A549/DDP 组、si NC 组、si SNHG15 组、 si SNHG15+inhibitor NC 组、si SNHG15+miR⁃483⁃3p inhibitor 组、si SNHG15+miR⁃483⁃3p inhibitor+si NC 组、 si SNHG15+miR⁃483⁃3p inhibitor+si DDIT4 组;蛋白印迹分析法检测各组细胞 E⁃cad、N⁃cad、DDIT4 蛋白表达 变化;双荧光素酶报告实验验证 miR⁃483⁃3p LncRNA SNHG15、DDIT4 的靶向关系。结果 A549 细胞相比,A549/DDP 细胞对 DDP IC50、LncRNA SNHG15、DDIT4 mRNA 与蛋白升高,miR⁃483⁃3p 水平降低(P < 0.05);A549/DDP 组细胞间黏附明显被破坏;si SNHG15 组、si SNHG15+miR⁃483⁃3p inhibitor+si DDIT4 组细 胞排列相对紧密,呈现上皮细胞特性;si SNHG15+miR⁃483⁃3p inhibitor组较si SNHG15组细胞排列松散、扩散细胞多;与 A549/DDP 组相比,si SNHG15 A549/DDP 细胞增殖抑制率、E⁃cad 表达升高,侵袭细胞数、迁移 细胞数、N⁃cad、DDIT4 表达降低(P < 0.05);与 si SNHG15 组相比,si SNHG15+miR⁃483⁃3p inhibitor A549/ DDP 细胞增殖抑制率、E⁃cad 表达降低,侵袭细胞数、迁移细胞数、N⁃cad、DDIT4 表达升高(P < 0.05)。结论 沉默 A549/DDP 细胞中 SNHG15 表达可通过调控 miR⁃483⁃3p/DDIT4,抑制 A549/DDP 细胞增殖、侵袭、迁移、 EMT,并降低A549/DDP 细胞对DDP 的耐药性。

关键词:

LncRNA SNHG15, miR?483?3p, DNA 损伤诱导转录物 4, 非小细胞肺癌, 顺铂, 上皮间质转化

Abstract:

Objective To investigate the impacts of LncRNA SNHG15 on cisplatin(DDP)sensitivity and epithelial⁃mesenchymal transition of non⁃small cell lung cancer(NSCLC)cells and the role of miR⁃483⁃3p/DNA damage⁃induced transcript 4(DDIT4)axis. Methods NSCLC cells(A549)and NSCLC drug⁃resistant cells(A549/ DDP)were cultured in vitro,the IC50 of the two cells to DDP and the mRNA and protein expression of LncRNA SN⁃ HG15,miR⁃483⁃3p and DDIT4 in the two cells were detected.A549/DDP cells were randomly divided into A549/ DDP group,si NC group,si SNHG15 group,si SNHG15+inhibitor NC group,si SNHG15+miR⁃483⁃3p inhibitor group,si SNHG15+miR⁃483⁃3p inhibitor+si NC group,and si SNHG15+miR⁃483⁃3p inhibitor+si DDIT4 group. Western blot was used to detect the changes of E⁃cad,N⁃cad and DDIT4 protein expression in each group anddual⁃ luciferase reporter assay was used to verify the targeting relationship of miR ⁃ 483⁃3p with LncRNA SNHG15 and DDIT4. Results Compared with A549 cells,the IC50,LncRNA SNHG15,DDIT4 mRNA and protein of DDP in A549/DDP cells increased,and the miR⁃483⁃3p level decreased(P < 0.05). In A549/DDP group,intercellular adhesion was significantly destroyed. The cells of si SNHG15 group and si SNHG15 + miR ⁃ 483 ⁃ 3p inhibitor + si DDIT4 group were relatively closely arranged,showing epithelial characteristics. The cells in the si SNHG15+miR⁃ 483⁃3p inhibitor group were loosely arranged and diffused as compared with the si SNHG15 group. Compared with A549/DDP group,the inhibition rate of A549/DDP cell proliferation and the expression of E ⁃ cad in si SNHG15group increased,while the number of invasive cells,the number of migrating cells,the expression of N⁃cad and DDIT4 decreased(P < 0.05). Compared with the si SNHG15 group,the proliferation inhibition rate and E ⁃cad expression of A549/DDP cells in the si SNHG15+miR⁃483⁃3p inhibitor group decreased,and the number of invasive cells,the number of migrating cells,N⁃cad and DDIT4 expression increased(P < 0.05). Conclusion Silencing the expression of SNHG15 in A549/DDP cells can inhibit the proliferation,invasion,migration and EMT of A549/ DDP cells,and reduce the drug resistance of A549/DDP cells to DDP by regulating miR⁃483⁃3p/DDIT4.

Key words:

"> LncRNA SNHG15, miR?483?3p, DNA damage?inducible transcript 4, non?small cell lung cancer, cisplatin, epithelial?mesenchymal transition