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环状 RNA circ_0000372 通过海绵吸附 miR-495 来提高 IL6 表达,促进结直肠癌细胞的增殖、迁移和侵袭。

Circular RNA circ_0000372 contributes to the proliferation, migration and invasion of colorectal cancer by elevating IL6 expression via sponging miR-495.

机构信息

Departments of Anorectal Surgery.

Acupuncture Physiotherapy, Jining No.1 People's Hospital, Jining City, Shandong Province, China.

出版信息

Anticancer Drugs. 2021 Mar 1;32(3):296-305. doi: 10.1097/CAD.0000000000001002.

DOI:10.1097/CAD.0000000000001002
PMID:33534412
Abstract

Circular RNAs are thought to play a vital function in the progression of various cancers, including colorectal cancer (CRC). However, the biological function and mechanism of circ_0000372 in CRC are still not clear. The expression of circ_0000372 and microRNA (miR)-495 was examined by quantitative real-time PCR. Cell proliferation was evaluated using cell counting kit 8 and colony formation assays. Further, cell migration and invasion were assessed using transwell assay. Additionally, western blot analysis was used to detect the expression of proteins associated with proliferation, metastasis, Janus kinase 2 (JAK2)/signal transducers and activators of transcription (STAT3) signaling pathway and interleukin 6 (IL6). Dual-luciferase reporter assay and RNA immunoprecipitation assay were employed to verify the interaction between miR-495 and circ_0000372 or IL6. Furthermore, the effect of circ_0000372 on CRC tumor growth in vivo was explored using the mice xenograft models. Circ_0000372 was markedly upregulated in CRC, and its high expression was associated with the poor prognosis of CRC patients. Silenced circ_0000372 was able to suppress CRC cell proliferation, migration and invasion in vitro and CRC tumor growth in vivo. Bioinformatics prediction and experimental verification proposed that circ_0000372 could sponge miR-495, and miR-495 could target IL6. Besides, the JAK2/STAT3 signaling pathway activation could be regulated by circ_0000372, miR-495 and IL6. Rescue assay results confirmed that the inhibition effect of circ_0000372 knockdown on the proliferation and metastasis of CRC could be reversed by miR-495 inhibitor or IL6 overexpression. In short, we concluded that circ_0000372 promoted CRC progression by regulating the miR-495/IL6 axis, suggesting that circ_0000372 could be used as a new prognostic biomarker and therapeutic target for CRC.

摘要

环状 RNA 被认为在包括结直肠癌(CRC)在内的多种癌症的进展中发挥着至关重要的作用。然而,circ_0000372 在 CRC 中的生物学功能和机制尚不清楚。通过实时定量 PCR 检测 circ_0000372 和 microRNA(miR)-495 的表达。使用细胞计数试剂盒 8 和集落形成测定法评估细胞增殖。此外,使用 Transwell 测定法评估细胞迁移和侵袭。另外,使用 Western blot 分析检测与增殖、转移、Janus 激酶 2(JAK2)/信号转导和转录激活因子 3(STAT3)信号通路和白细胞介素 6(IL6)相关的蛋白表达。双荧光素酶报告基因检测和 RNA 免疫沉淀检测用于验证 miR-495 与 circ_0000372 或 IL6 之间的相互作用。此外,使用小鼠异种移植模型探讨了 circ_0000372 对 CRC 肿瘤生长的体内影响。circ_0000372 在 CRC 中明显上调,其高表达与 CRC 患者的不良预后相关。沉默 circ_0000372 能够抑制 CRC 细胞在体外的增殖、迁移和侵袭以及体内的 CRC 肿瘤生长。生物信息学预测和实验验证表明,circ_0000372 可以海绵 miR-495,miR-495 可以靶向 IL6。此外,circ_0000372、miR-495 和 IL6 可以调节 JAK2/STAT3 信号通路的激活。挽救实验结果证实,circ_0000372 敲低对 CRC 增殖和转移的抑制作用可以通过 miR-495 抑制剂或 IL6 过表达逆转。总之,我们得出结论,circ_0000372 通过调节 miR-495/IL6 轴促进 CRC 的进展,表明 circ_0000372 可以作为 CRC 的新预后生物标志物和治疗靶点。

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