LncRNA UCA1对肺癌细胞放射敏感性的影响及其机制

中华放射肿瘤学杂志 Pub Date : 2020-04-15 DOI:10.3760/CMA.J.CN113030-20190618-00008

Cheng Wang, Zongwen Liu, Ge Hou, Jun Yang, Yang-yang Huang

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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. 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引用次数: 1

摘要

目的探讨长链非编码RNA (LncRNA) UCA1对肺癌细胞增殖、凋亡及放射敏感性的影响，并探讨其作用机制。方法采用qRT-PCR检测肺癌细胞A549、H1299和正常人肺细胞HBE中UCA1和miR-513a-5p的表达。si-con组(转染的si-con)、si-UCA1组(转染的si-UCA1)、miR-513a-5p组(转染的miR-513a-5p模拟物)、miR-NC组(转染的miR-NC)、IR+ si-con组(转染的si-con并照射)、IR+ si-UCA1组(转染的miR-NC并照射)、IR+ miR-513a-5p组(转染的miR-513a-5p模拟物并照射)、IR+ miR-NC组(转染的miR-NC并照射)、IR+ si-UCA1+抗miR-513a-5p组(转染的miR-NC并照射)、IR+ miR-NC组(转染的miR-NC并照射)、IR+ si-UCA1+抗miR-513a-5p组(共转染的si-UCA1、通过脂质体法将anti-miR-513a-5p和辐照后的anti-miR-513a-5p)转染到A549和H1299细胞中，然后对部分组细胞进行4Gy辐照。MTT法检测各组细胞增殖情况。通过克隆形成试验评估敏感性增强率。流式细胞术检测各组细胞凋亡情况。采用双荧光素基因检测法检测各组的荧光活性。结果与人正常肺细胞HBE相比，肺癌细胞A549和H1299中UCA1表达水平显著上调(P<0.05)， miR-513a-5p表达水平显著下调(P<0.05)。抑制UCA1和过表达miR-513a-5p可显著抑制细胞增殖，促进细胞凋亡，提高A549和H1299对辐射暴露的敏感性(敏感性增强比分别为1.897、2.146和1.615、1.872)。miR-513a-5p可以抑制野生型UCA1细胞的荧光活性，UCA1可以负向调节miR-513a-5p的表达。抑制miR-513a-5p可逆转抑制UCA1对肺癌细胞放射敏感性的增强作用。结论抑制LncRNA UCA1可增强肺癌细胞对辐射暴露的敏感性。其机制可能与miR-513a-5p的靶向抑制有关。关键词:UCA1基因;mir - 513 - 5 - p基因;辐射敏感度;肺癌细胞系

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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. Key words: UCA1 gene; miR-513a-5p gene; Radiosensitivity; Lung cancer cell line

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来源期刊

中华放射肿瘤学杂志

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6375

期刊介绍： The Chinese Journal of Radiation Oncology is a national academic journal sponsored by the Chinese Medical Association. It was founded in 1992 and the title was written by Chen Minzhang, the former Minister of Health. Its predecessor was the Chinese Journal of Radiation Oncology, which was founded in 1987. The journal is an authoritative journal in the field of radiation oncology in my country. It focuses on clinical tumor radiotherapy, tumor radiation physics, tumor radiation biology, and thermal therapy. Its main readers are middle and senior clinical doctors and scientific researchers. It is now a monthly journal with a large 16-page format and 80 pages of text. For many years, it has adhered to the principle of combining theory with practice and combining improvement with popularization. It now has columns such as monographs, head and neck tumors (monographs), chest tumors (monographs), abdominal tumors (monographs), physics, technology, biology (monographs), reviews, and investigations and research.