Department of General Surgery, the First Affiliated Hospital of Jinzhou Medical University, Jinzhou, P.R. China.
Radiology, the First Affiliated Hospital of Jinzhou Medical University, Jinzhou, P.R. China.
Cancer Med. 2020 Sep;9(18):6710-6725. doi: 10.1002/cam4.3332. Epub 2020 Jul 24.
In our study, has_circRNA_102209 was the most elevated regulator in colorectal cancer (CRC) tissues according to circRNA array data. The levels of hsa_circRNA_102209 in CRC specimens and cells, as well as its effects on CRC cells were investigated. The expression of hsa_circRNA_102209 in CRC and paired non-cancerous samples, human CRC, and normal colonic epithelial cells were examined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Cells with hsa_circRNA_102209 knockdown were established using lentiviral vectors. Cell proliferative ability was evaluated using CCK-8 assay; cell migrative/invasive activities were determined using wound healing/Transwell assay. Cell cycle arrest and apoptosis were assessed by flow cytometry; apoptosis, and EMT markers were examined using RT-qPCR and western blotting. Tumor development and levels of associated proteins were determined in hsa_circRNA_102209 knockdown mice. Our results revealed that expression of hsa_circRNA_102209 was remarkably increased in CRC tissues, where the levels of miR-761 were notably reduced (P < .05). Additionally, the levels of hsa_circRNA_102209 were associated with histology grade and occurrence of liver metastasis in CRC patients, and the expression of hsa_circRNA_102209 and miR-761 were negatively correlated (P < .05). Moreover, hsa_circRNA_102209 was upregulated in CRC cells compared with normal colonic epithelial cells. Knockdown of hsa_circRNA_102209 notably inhibited the proliferation, migration, invasion, and EMT of CRC cells (P < .05), whereas cell cycle arrest at G0/G1 phase and apoptosis were enhanced (P < .05). Furthermore, miR-761/Ras and Rab interactor 1 (RIN1) axis was the putative target of hsa_circRNA_102209 in CRC and involved in hsa_circRNA_102209-modulated growth and metastasis of CRC cells (P < .05). Knockdown of hsa_circRNA_102209 also remarkably suppressed tumor growth in vivo (P < .05). In summary, our data revealed that the expression of hsa_circRNA_102209 was elevated in CRC samples and cells. Furthermore, hsa_circRNA_102209 could promote the progression of CRC through miR-761/RIN1 axis. More importantly, hsa_circRNA_102209/miR-761/RIN1 signaling may be a novel therapeutic target for the treatment of CRC patients.
在我们的研究中，根据 circRNA 阵列数据，hsa_circRNA_102209 是结直肠癌 (CRC) 组织中升高最多的调节因子。研究了 hsa_circRNA_102209 在 CRC 标本和细胞中的水平及其对 CRC 细胞的影响。使用逆转录-定量聚合酶链反应 (RT-qPCR) 检测 hsa_circRNA_102209 在 CRC 和配对非癌组织、人 CRC 和正常结肠上皮细胞中的表达。使用慢病毒载体建立 hsa_circRNA_102209 敲低细胞。使用 CCK-8 测定法评估细胞增殖能力；通过划痕愈合/Transwell 测定法测定细胞迁移/侵袭活性。通过流式细胞术评估细胞周期停滞和细胞凋亡；使用 RT-qPCR 和 Western blot 检测细胞凋亡和 EMT 标志物。在 hsa_circRNA_102209 敲低小鼠中确定肿瘤发生和相关蛋白水平。我们的结果表明，hsa_circRNA_102209 在 CRC 组织中表达显著增加，miR-761 的水平明显降低（P<.05）。此外，hsa_circRNA_102209 的水平与 CRC 患者的组织学分级和肝转移的发生有关，并且 hsa_circRNA_102209 和 miR-761 的表达呈负相关（P<.05）。此外，hsa_circRNA_102209 在 CRC 细胞中上调，而在正常结肠上皮细胞中下调。hsa_circRNA_102209 敲低显著抑制 CRC 细胞的增殖、迁移、侵袭和 EMT（P<.05），而细胞周期停滞在 G0/G1 期并促进细胞凋亡（P<.05）。此外，miR-761/Ras 和 Rab 相互作用蛋白 1 (RIN1) 轴是 hsa_circRNA_102209 在 CRC 中的推定靶标，并参与 hsa_circRNA_102209 调节的 CRC 细胞生长和转移（P<.05）。hsa_circRNA_102209 敲低也显著抑制体内肿瘤生长（P<.05）。总之，我们的数据表明 hsa_circRNA_102209 在 CRC 样本和细胞中的表达升高。此外，hsa_circRNA_102209 可以通过 miR-761/RIN1 轴促进 CRC 的进展。更重要的是，hsa_circRNA_102209/miR-761/RIN1 信号可能是治疗 CRC 患者的一种新的治疗靶点。
