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Silencing LncRNA OIP5-AS1 increases radiosensitivity of non-small cell lung cancer A549 cell line by up-regulating miR-34c-5p expression
Mao Kai1, Ding Xiaohua2, Wu Liping3, Mao Yujing4, Zhang Liguo1, Li Jun1, Lu Jiang1
1Department of Thoracic Oncology 2Department of Radiotherapy, Xinxiang Central Hospital, Xinxiang 453000 China 3Sanquan Medical College, College of Inspection and Imaging, Xinxiang 453003, China 4First Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, China
AbstractObjective 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.
Mao Kai,Ding Xiaohua,Wu Liping et al. Silencing LncRNA OIP5-AS1 increases radiosensitivity of non-small cell lung cancer A549 cell line by up-regulating miR-34c-5p expression[J]. Chinese Journal of Radiation Oncology, 2020, 29(1): 57-60.
Mao Kai,Ding Xiaohua,Wu Liping et al. Silencing LncRNA OIP5-AS1 increases radiosensitivity of non-small cell lung cancer A549 cell line by up-regulating miR-34c-5p expression[J]. Chinese Journal of Radiation Oncology, 2020, 29(1): 57-60.
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2020, Vol. 29 
