Silencing LncRNA OIP5-AS1 increases radiosensitivity of non-small cell lung cancer A549 cell line by up-regulating miR-34c-5p expression
10.3760/cma.j.issn.1004-4221.2020.01.013
- VernacularTitle: 沉默LncRNA OIP5-AS1通过上调miR-34c-5p表达增加A549R细胞放射敏感性
- Author:
Kai MAO
1
;
Xiaohua DING
2
;
Liping WU
3
;
Yujing MAO
4
;
Liguo ZHANG
1
;
Jun LI
1
;
Jiang LU
1
Author Information
1. Department of Thoracic Oncology, Xinxiang Central Hospital, Xinxiang 453000 China
2. Department of Radiotherapy, Xinxiang Central Hospital, Xinxiang 453000 China
3. Sanquan Medical College, College of Inspection and Imaging, Xinxiang 453003, China
4. First Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, China
- Publication Type:Journal Article
- Keywords:
A549 cell line;
A549R cell line;
OIP5-AS1 gene;
miR-34c-5p gene;
radiosensitivity
- From:
Chinese Journal of Radiation Oncology
2020;29(1):57-60
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of LncRNA OIP5-AS1 on radiosensitivity of non-small cell lung cancer (NSCLC) cells and its mechanism.
Methods:The radiation-resistant cell A549R was established by using A549 cells irradiated by X-ray 6Gy in 5 fractions. The expression levels of OIP5-AS1 and miR-34c-5p in A549 and A549R cells were detected by qRT-PCR. OIP5-AS1 inhibitor or miR-34c-5p mimetic was transfected into A549R cells, or OIP5-AS1 overexpression plasmid was transfected into A549 cells. Cell apoptosis was detected by flow cytometry. Cell radiosensitivity was analyzed by colony formation assay. The expression levels of p-Chk2 and p-ATM proteins were measured by Western blot. Dual luciferase assay was adopted to verify the relationship between OIP5-AS1 and miR-34c-5p.
Results:Compared with A549 cells, the expression of OIP5-AS1 was significantly up-regulated in A549R cells (1.97±0.11 vs.1.01±0.05, P<0.05), whereas the expression of miR-34c-5p was remarkably down-regulated (0.43±0.02 vs.1.02±0.06, P<0.05). The expression levels of p-Chk2 and p-ATM proteins in A549R cells in the silencing OIP5-AS1+ 6Gy group were significantly lower (0.43±0.03 vs.1.39±0.15, 0.51±0.0 5 vs.1.21± 0.11, both P<0.05), whereas the apoptotic rate was significantly higher than those in the silencing control+ 6Gy group [(13.29±1.25)% vs. (28.47±2.31)%, P<0.05)]. The expression levels of p-Chk2 and p-ATM proteins in A549 cells in overexpressing OIP5-AS1+ 6Gy group were significantly higher than those in overexpression control+ 6Gy group (1.23±0.13 vs.0.75±0.06, 1.08±0.11 vs.0.59±0.04, both P<0.05). Inhibiting miR-34c-5p expression reversed the effect of silencing OIP5-AS1 on survival fraction of A549R cells (SER=1.42). OIP5-AS1 negatively regulated the expression of miR-34c-5p.
Conclusion:Silencing OIP5-AS1 enhances the radiosensitivity of radiation-resistant A549 cells by up-regulating the expression of miR-34c-5p, providing a potential target for radiotherapy of NSCLC cells.
