Department of Respiratory and Critical Care Medicine, Shanxi Provincial People's Hospital, Taiyuan, China.
Eur Rev Med Pharmacol Sci. 2020 Apr;24(8):4271-4280. doi: 10.26355/eurrev_202004_21007.
It has been demonstrated that circular RNA (circRNA) plays an important regulatory role in a series of diseases. The purpose of this study is to investigate the expression of circRNA_001010 and its facilitating effects on proliferation and invasion of non-small cell lung cancer (NSCLC) by regulating oncogene CDK4 through sponging with miR-5112.
qRT-PCR was performed to detect the expressions of circRNA_001010 and CDK4 in human NSCLC tissues and cells. Cell Counting Kit-8 (CCK-8) assay was performed to evaluate the A549 cells proliferation and transwell assay was performed to evaluate the A549 cells migration. Correlation analysis between circRNA_001010 and miR-5112 was detected by statistical analysis. Bioinformatics prediction was made to detect the binding site of GTL and miR-5112 and Luciferase activity was conducted to investigate the interaction between circRNA_001010 and miR-5112. Furthermore, we cloned the mice CDK4 3'-UTR into the Luciferase reporter vector and constructed miR-5112 binding mutants to validate the inhibited modulation of miR-5112 to the CDK4 expression.
Results showed that the expressions of circRNA_001010 and CDK4 were upregulated in human NSCLC tissues and cells. qRT-PCR and CCK-8 assay showed that circRNA_001010 expression is associated with the proliferation of NSCLC cells, and that upregulated circRNA_001010 contributed to cell proliferation of A549. Transwell assay showed that circRNA_001010 was associated with the migration ability of tumor cells, and that increased expression of circRNA_001010 promoted the migration and invasion of NSCLC cells. The bioinformatics prediction and Luciferase assay demonstrated that by sponging with miR-5112, circRNA_001010 can serve as a ceRNA for miR-5112 to further regulate the expression of CDK4.
For the first time, we found that circRNA_001010 was upregulated in human NSCLC patients, which could accelerate tumor proliferation, migration and invasion as a molecular sponge by modulating the inhibitory effect of miR-5112 on oncogene CDK4.
已有研究表明环状 RNA（circRNA）在一系列疾病中发挥重要的调控作用。本研究旨在通过circRNA_001010 与 miR-5112 结合，海绵吸附作用调控癌基因 CDK4，探讨 circRNA_001010 的表达及其对非小细胞肺癌（NSCLC）增殖和侵袭的促进作用。
采用 qRT-PCR 检测人 NSCLC 组织和细胞中 circRNA_001010 和 CDK4 的表达。采用细胞计数试剂盒-8（CCK-8）法评估 A549 细胞增殖，Transwell 法评估 A549 细胞迁移。通过统计学分析检测 circRNA_001010 与 miR-5112 的相关性。采用生物信息学预测 GTL 与 miR-5112 的结合位点，并进行荧光素酶活性检测以研究 circRNA_001010 与 miR-5112 之间的相互作用。此外，我们将小鼠 CDK4 3'-UTR 克隆到荧光素酶报告载体中，并构建 miR-5112 结合突变体，以验证 miR-5112 对 CDK4 表达的抑制调节作用。
结果表明，circRNA_001010 和 CDK4 在人 NSCLC 组织和细胞中表达上调。qRT-PCR 和 CCK-8 检测结果显示，circRNA_001010 的表达与 NSCLC 细胞的增殖有关，上调的 circRNA_001010 促进了 A549 细胞的增殖。Transwell 检测结果显示，circRNA_001010 与肿瘤细胞的迁移能力有关，上调 circRNA_001010 促进了 NSCLC 细胞的迁移和侵袭。生物信息学预测和荧光素酶检测表明，circRNA_001010 通过与 miR-5112 结合，作为 miR-5112 的 ceRNA 进一步调节 CDK4 的表达。
本研究首次发现 circRNA_001010 在人 NSCLC 患者中上调，可作为分子海绵通过调节 miR-5112 对癌基因 CDK4 的抑制作用，加速肿瘤增殖、迁移和侵袭。
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