LncRNA UCA1对肺癌细胞放射敏感性影响及机制研究

目的 研究长链非编码RNA (LncRNA) UCA1对肺癌细胞增殖、凋亡及放射敏感性影响及其机制。方法 运用qRT-PCR法检测肺癌细胞A549、H1299和人正常肺细胞HBE中UCA1、miR-513a-5p表达。将si-con组(转染si-con)、si-UCA1组(转染si-UCA1)、miR-513a-5p组(转染miR-513a-5p mimics)、miR-NC组(转染miR-NC)、IR+si-con组(转染si-con+照射)、IR+si-UCA1组(转染miR-NC+照射)、IR+miR-513a-5p组(转染miR-513a-5p mimics+照射)、IR+miR-NC组(转染miR-NC+照射)、IR+si-UCA1+anti-miR-513a-5p组(共转染si-UCA1和anti-miR-513a-5p+照射)均用脂质体法转染至A549、H1299细胞,然后部分组进行4Gy照射。MTT法检测各组细胞增殖,克隆形成实验检测细胞增敏比,流式细胞术检测各组细胞凋亡,双荧光素没报告基因检测实验检测各组细胞的荧光活性。结果 与HBE细胞相比,A549、H1299细胞中UCA1表达显著升高(P<0.05),miR-513a-5p表达显著降低(P<0.05)。抑制UCA1、过表达miR-513a-5p均可明显抑制A549、H1299细胞增殖、促进凋亡、提高放射敏感性(放射增敏比为1.897、2.146和1.615、1.872)。miR-513a-5p可抑制野生型UCA1细胞的荧光活性,且UCA1可负向调控miR-513a-5p的表达。抑制miR-513a-5p可逆转抑制UCA1对细胞的放射敏感性的增强作用。结论 抑制LncRNA UCA1可增强放射对肺癌细胞敏感性,其机制可能与靶向抑制miR-513a-5p有关。

Effect of LncRNA UCA1 on radiosensitivity of lung cancer cells and its mechanism

Objective To evaluate the effect of long-chain non-coding RNA (LncRNA) UCA1 on the proliferation,apoptosis and radiosensitivity of lung cancer cell and to explore the underlying mechanism. Methods qRT-PCR was used to detect the expression of UCA1 and miR-513a-5p in lung cancer cell A549,H1299 and normal human lung cell HBE. The si-con group (transfected si-con),si-UCA1 group (transfected si-UCA1),miR-513a-5p group (transfected miR-513a-5p mimics),miR-NC group (transfected miR-NC),IR+si-con group (transfected si-con,and irradiated),IR+si-UCA1 group (transfected miR-NC and irradiated),IR+miR-513a-5p group (transfected miR-513a-5p mimics and irradiated),IR+miR-NC group (transfected miR-NC and irradiated),IR+si-UCA1+anti-miR-513a-5p group (co-transfected si-UCA1, anti-miR-513a-5p and irradiated) were transfected into the A549 and H1299 cells by liposome method,and then the cells in certain groups were subject to 4Gy irradiation. The cell proliferation of each group was detected by MTT assay. The sensitivity enhancement ratio was assessed by clone formation assay. The cell apoptosis of each group was detected by flow cytometry. The fluorescence activity of each group was detected by dual-fluorescein gene detection assay. Results Compared with human normal lung cell HBE,the expression levels of UCA1 were significantly up-regulated in the lung cancer cell A549 and H1299(both P<0.05),whereas those of miR-513a-5p were significantly down-regulated (both P<0.05). Inhibition of UCA1 and overexpression of miR-513a-5p significantly inhibited cell proliferation,promoted cell apoptosis and increased the sensitivity of radiation exposure of A549 and H1299(sensitivity enhancement ratio=1.897, 2.146 and 1.615,1.872). miR-513a-5p could suppress the fluorescence activity of wild-type UCA1 cells,and UCA1 could negatively regulate the expression of miR-513a-5p. Inhibition of miR-513a-5p could reverse the enhancement effect of inhibiting UCA1 upon the radiosensitivity of lung cancer cells. Conclusions Inhibition of LncRNA UCA1 can enhance the sensitivity of radiation exposure to lung cancer cells. The mechanism may be related to the targeted inhibition of miR-513a-5p.