沉默 circPVT1 通过海绵吸附 microRNA-1208 增强非小细胞肺癌的放射敏感性。
Silencing circPVT1 enhances radiosensitivity in non-small cell lung cancer by sponging microRNA-1208.
机构信息
Department of Oncology, The First People's Hospital of Qujing/The Qujing Affiliated Hospital of Kunming Medical University, Qujing, Yunnan, China.
Department of Thoracic Surgery, The First People's Hospital of Qujing/The Qujing Affiliated Hospital of Kunming Medical University, Qujing, Yunnan, China.
出版信息
Cancer Biomark. 2021;31(3):263-279. doi: 10.3233/CBM-203252.
BACKGROUND
Radiotherapy is one of main useful therapies in non-small cell lung cancer (NSCLC). Nevertheless, the underlying mechanism between NSCLC cell radiosensitivity and effective treatment remains unclear.
OBJECTIVE
The aim is to explore the relationship between circular (circ) RNA and NSCLC cell radiosensitivity.
METHODS
CircRNA plasmacytoma variant translocation 1 (PVT1) and microRNA (miR)-1208 expression in NSCLC cells were assessed using quantitative reverse transcriptase PCR (qRT-PCR). NSCLC cells were transfected with si-PVT1 or miR-1208 inhibitor and then exposed to irradiation. Cellular biology behaviors were detected using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Terminal deoxynucleotidyl transferase dUTP Nick-End Labeling (TUNEL), colony formation, invasion and western blot. Additionally, binding between circPVT1 and miR-1208 was testified by dual-luciferase reporter and RIP assay.
RESULTS
CircPVT1 was upregulated in NSCLC cells after irradiation treatment. Silencing circPVT1 induced inhibition of NSCLC cell growth and invasion, accompanied by cell apoptosis and γ-H2AX expression. Moreover, NSCLC cell proliferation and invasion was further inhibited by irradiation treatment in circPVT1-silenced cells, indicating a strong radiosensitivity of NSCLC cells. CircPVT1 functions as a competing endogenous RNA of miR-1208. Silencing miR-1208 reversed NSCLC cell sensitivity response to irradiation and activated PI3K/AKT/mTOR pathway in circPVT1-silenced cells.
CONCLUSIONS
Silencing circPVT1 enhanced radiosensitivity of NSCLC cells by sponging miR-1208.
背景
放射治疗是非小细胞肺癌（NSCLC）的主要有效治疗方法之一。然而，NSCLC 细胞放射敏感性与有效治疗之间的潜在机制仍不清楚。
目的
旨在探讨环状 RNA 与 NSCLC 细胞放射敏感性之间的关系。
方法
采用实时定量逆转录 PCR（qRT-PCR）检测 NSCLC 细胞中浆细胞瘤变异易位 1（PVT1）和 microRNA（miR）-1208 的表达。用 si-PVT1 或 miR-1208 抑制剂转染 NSCLC 细胞，然后进行照射。采用 3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）、末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）、集落形成、侵袭和 Western blot 检测细胞生物学行为。此外，通过双荧光素酶报告和 RIP 测定验证 circPVT1 与 miR-1208 之间的结合。
结果
照射后 NSCLC 细胞中 circPVT1 上调。沉默 circPVT1 诱导 NSCLC 细胞生长和侵袭抑制，伴有细胞凋亡和 γ-H2AX 表达。此外，circPVT1 沉默细胞经照射处理后进一步抑制 NSCLC 细胞增殖和侵袭，表明 NSCLC 细胞具有较强的放射敏感性。CircPVT1 作为 miR-1208 的竞争性内源性 RNA。沉默 miR-1208 逆转了 circPVT1 沉默细胞对照射的敏感性反应，并激活了 circPVT1 沉默细胞中的 PI3K/AKT/mTOR 通路。
结论
沉默 circPVT1 通过海绵吸附 miR-1208 增强 NSCLC 细胞的放射敏感性。
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