Department of Oncology, Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine, Nanjing, China.
First Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, China.
J Clin Lab Anal. 2022 May;36(5):e24393. doi: 10.1002/jcla.24393. Epub 2022 Apr 4.
Caudatin is extracted from radix cynanchi bungei and has an inhibitory effect on cancer progression. The study aims to reveal the impacts of hsa_circ_0060927 on Caudatin-mediated colorectal cancer (CRC) development and the underneath mechanism.
The expression levels of hsa_circ_0060927, microRNA-421 (miR-421) and miR-195-5p were detected by quantitative real-time reverse transcription-polymerase chain reaction. The protein expression was analyzed by Western blot or immunohistochemistry assay. Cell viability and proliferation were analyzed by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide or 5-Ethynyl-29-deoxyuridine assay. Cell apoptosis was quantified by flow cytometry analysis. Cell migration and invasion were investigated by transwell assay. The putative associations among hsa_circ_0060927, miR-421 and miR-195-5p were predicted by the starbase online database, and identified by dual-luciferase reporter, RNA pull-down and RNA immunoprecipitation (RIP) assays. The impacts of Caudatin treatment on tumor growth in vivo were revealed by a xenograft tumor model assay.
Hsa_circ_0060927 expression was significantly upregulated, whereas miR-421 and miR-195-5p were downregulated in CRC tissues and cells compared with control groups. Hsa_circ_0060927 expression was closely associated with lymph node metastasis and tumor-node-metastasis stage. Caudatin treatment significantly decreased hsa_circ_0060927 expression but increased miR-421 and miR-195-5p expression. Caudatin exposure suppressed CRC cell proliferation, migration and invasion, and induced cell apoptosis; however, hsa_circ_0060927 overexpression hindered these impacts. Additionally, hsa_circ_0060927 was associated with miR-421/miR-195-5p. Depletion of miR-421 or miR-195-5p attenuated the influences of hsa_circ_0060927 silencing on CRC development. Furthermore, Caudatin treatment repressed tumor growth in vivo.
Caudatin inhibited CRC cell malignancy through the hsa_circ_0060927/miR-421/miR-195-5p pathway, which provided a potential therapeutic agent for CRC.
钩藤碱是从徐长卿中提取的，对癌症的发展具有抑制作用。本研究旨在揭示 hsa_circ_0060927 对 Caudatin 介导的结直肠癌（CRC）发展的影响及其潜在机制。
采用实时定量逆转录聚合酶链反应检测 hsa_circ_0060927、微小 RNA-421（miR-421）和 miR-195-5p 的表达水平。通过 Western blot 或免疫组织化学检测蛋白质表达。采用 3-(4,5)-二甲基噻唑（-z-y1)-3,5-二苯基四唑溴盐或 5-乙炔基-29-脱氧尿苷测定细胞活力和增殖。通过流式细胞术分析定量细胞凋亡。通过 Transwell 测定研究细胞迁移和侵袭。通过 starbase 在线数据库预测 hsa_circ_0060927、miR-421 和 miR-195-5p 之间的可能关联，并通过双荧光素酶报告、RNA 下拉和 RNA 免疫沉淀（RIP）测定进行鉴定。通过异种移植肿瘤模型试验揭示 Caudatin 处理对体内肿瘤生长的影响。
与对照组相比，CRC 组织和细胞中 hsa_circ_0060927 表达显著上调，而 miR-421 和 miR-195-5p 表达下调。hsa_circ_0060927 表达与淋巴结转移和肿瘤-淋巴结-转移分期密切相关。Caudatin 处理显著降低 hsa_circ_0060927 表达，但增加 miR-421 和 miR-195-5p 表达。Caudatin 暴露抑制 CRC 细胞增殖、迁移和侵袭，并诱导细胞凋亡；然而，hsa_circ_0060927 的过表达阻碍了这些影响。此外，hsa_circ_0060927 与 miR-421/miR-195-5p 相关。miR-421 或 miR-195-5p 的耗竭减弱了 hsa_circ_0060927 沉默对 CRC 发展的影响。此外，Caudatin 处理抑制体内肿瘤生长。
Caudatin 通过 hsa_circ_0060927/miR-421/miR-195-5p 通路抑制 CRC 细胞恶性，为 CRC 提供了一种潜在的治疗药物。
