长链非编码 RNA ANRIL 通过海绵吸附 miR-181a-5p 负调控壳寡糖诱导的结肠癌细胞放射敏感性。
LncRNA ANRIL negatively regulated chitooligosaccharide-induced radiosensitivity in colon cancer cells by sponging miR-181a-5p.
机构信息
Department of Radiology, The First Affiliated Hospital of Soochow University, Affiliated Hospital of Nantong University, Department of Nuclear Medicine, China.
Department of Gastrointestinal Surgery, Affiliated Hospital of Nantong University, China.
出版信息
Adv Clin Exp Med. 2021 Jan;30(1):55-65. doi: 10.17219/acem/128370.
BACKGROUND
The radiosensitivity of colon cancer cells can be regulated by noncoding RNAs.
OBJECTIVES
In this study, the lncRNA antisense non-coding RNA in the INK4 locus (ANRIL) was selected to analyze its regulatory role in chitooligosaccharides (COS)-related radiosensitivity in colon cancer cells.
MATERIAL AND METHODS
The ANRIL expression in colon cancer cell lines was examined using real-time quantitative polymerase chain reaction (RT-qPCR), based on which we selected the cell line that presented the highest expression of ANRIL for radiosensitivity research. The cells were exposed to X-rays (0 Gy, 2 Gy, 4 Gy, and 6 Gy) and evaluated for changes in ANRIL and miR-181a-5p expression using RT-qPCR. Cell viability was evaluated using the CCK8 method, while apoptosis was detected with flow cytometry assays. Dual luciferase assays validated the binding between ANRIL and miR-181a-5p. The cell survival rates after differential COS treatments (0 mg/mL, 1.0 mg/mL, 2.0 mg/mL, 3.0 mg/mL, 4.0 mg/mL, and 5.0 mg/mL) were rated using CCK8 assay. The cells with the strongest dosage of COS (5.0 mg/mL) were selected to further investigate the role of ANRIL/miR-181a-5p in modulating the radiosensitivity observed with CCK-8 and flow cytometry assays.
RESULTS
The ANRIL was highly expressed in colon cancer cells lines, especially in the SW480 cell line. Irradiation significantly decreased cell viability and ANRIL expression in a dose-dependent manner. The overexpression of ANRIL reduced the cell apoptosis rate after irradiation. MiR-181a-5p directly bound to ANRIL and was upregulated by irradiation in a dose-dependent manner. The suppression of miR-181a-5p decreased cell apoptosis. The COS treatment notably downregulated cell survival and promoted apoptosis in cells exposed to irradiation. The overexpression of ANRIL partially reversed COS-induced apoptosis and the inhibition rate; the upregulation of miR-181a-5p could counteract the impact of ANRIL regulation in cells.
CONCLUSIONS
The ANRIL negatively regulated radiosensitivity induced by COS in colon cancer cells by sponging miR-181a-5p.
背景
非编码 RNA 可调节结肠癌细胞的放射敏感性。
目的
本研究选择长链非编码 RNA 反义非编码 RNA 位于 INK4 基因座（ANRIL），分析其在壳寡糖（COS）相关结肠癌细胞放射敏感性中的调控作用。
材料与方法
采用实时定量聚合酶链反应（RT-qPCR）检测结肠癌细胞系中 ANRIL 的表达，在此基础上，选择 ANRIL 表达最高的细胞系进行放射敏感性研究。用 X 射线（0 Gy、2 Gy、4 Gy 和 6 Gy）照射细胞，用 RT-qPCR 检测 ANRIL 和 miR-181a-5p 表达的变化。用 CCK8 法评估细胞活力，用流式细胞术检测细胞凋亡。双荧光素酶实验验证 ANRIL 与 miR-181a-5p 的结合。用 CCK8 法评价不同 COS 处理（0 mg/ml、1.0 mg/ml、2.0 mg/ml、3.0 mg/ml、4.0 mg/ml 和 5.0 mg/ml）后细胞存活率。选择 COS 最强剂量（5.0 mg/ml）的细胞进一步研究 ANRIL/miR-181a-5p 在调节 CCK-8 和流式细胞术观察到的放射敏感性中的作用。
结果
ANRIL 在结肠癌细胞系中高表达，尤其在 SW480 细胞系中表达最高。照射呈剂量依赖性显著降低细胞活力和 ANRIL 表达。过表达 ANRIL 降低了照射后细胞凋亡率。miR-181a-5p 直接与 ANRIL 结合，并呈剂量依赖性被照射上调。抑制 miR-181a-5p 降低了细胞凋亡率。COS 处理明显降低了照射细胞的细胞存活率并促进了细胞凋亡。过表达 ANRIL 部分逆转了 COS 诱导的细胞凋亡和抑制率；上调 miR-181a-5p 可逆转 ANRIL 对细胞的调节作用。
结论
ANRIL 通过海绵吸附 miR-181a-5p 负调控 COS 诱导的结肠癌细胞放射敏感性。
Zotero 插件