Yang Tao, Sun Jufeng, Wang Wei, Li Dongsheng, Yang Xianxu, Jia Ang, Ma Yinda, Fan Zhongkai
The First Affiliated Hospital of Jinan University, Jinan University, Guangzhou, 510000, China.
Department of General Surgery, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, 121010, China.
Mol Cell Biochem. 2022 Dec;477(12):2751-2760. doi: 10.1007/s11010-022-04458-5. Epub 2022 May 26.
Colorectal cancer (CRC) remains a malignancy tumor with high metastasis and poor prognosis. We aimed to explore the effect of circular RNA (circRNA) hsa_circ_0006732 in the progression of CRC. Hsa_circ_0006732 expression in CRC tissues and cell lines were detected using qRT-PCR. The relationship between hsa_circ_0006732 expression and clinicopathologic characteristics of patients with CRC was analyzed. Loss-of-function assay was conducted to determine the regulatory effect of hsa_circ_0006732 on CRC cell proliferation, migration and invasion by using the CCK-8, wound-healing assay and transwell assays. Protein expression changes on epithelial mesenchymal transition (EMT)-related factors were detected by western blotting. The downstream signaling pathway was investigated by bioinformatics, dual-luciferase reporter assay. Rescue assay was further examined for prediction validation. It was found that hsa_circ_0006732 was highly expressed in CRC tissues and cell lines. Downregulation of hsa_circ_0006732 suppressed the proliferation, migration, invasion and EMT of CRC cells. Further mechanistic investigations proved that hsa_circ_0006732 functioned as a competitive endogenous RNA (ceRNA) by directly sponging of miR-127-3p, which further affected the expression of Ras-related protein Rab-3D (Rab3D). Taken together, these findings indicated that hsa_circ_0006732 might be an oncogene in CRC through the regulation of the miR-127-5p/RAB3D axis. Thus, hsa_circ_0006732 might serve as a potential therapeutic target for the treatment of CRC.
结直肠癌(CRC)仍然是一种具有高转移性和预后不良的恶性肿瘤。我们旨在探讨环状RNA(circRNA)hsa_circ_0006732在CRC进展中的作用。采用qRT-PCR检测CRC组织和细胞系中hsa_circ_0006732的表达。分析hsa_circ_0006732表达与CRC患者临床病理特征之间的关系。通过CCK-8、伤口愈合试验和Transwell试验进行功能缺失试验,以确定hsa_circ_0006732对CRC细胞增殖、迁移和侵袭的调节作用。通过蛋白质免疫印迹法检测上皮-间质转化(EMT)相关因子的蛋白质表达变化。通过生物信息学、双荧光素酶报告基因试验研究下游信号通路。进一步进行挽救试验以验证预测结果。结果发现,hsa_circ_0006732在CRC组织和细胞系中高表达。下调hsa_circ_0006732可抑制CRC细胞的增殖、迁移、侵袭和EMT。进一步的机制研究证明,hsa_circ_0006732通过直接海绵吸附miR-127-3p发挥竞争性内源性RNA(ceRNA)的作用,进而影响Ras相关蛋白Rab-3D(Rab3D)的表达。综上所述,这些发现表明hsa_circ_0006732可能通过调节miR-127-5p/RAB3D轴成为CRC中的一个癌基因。因此,hsa_circ_0006732可能成为治疗CRC的潜在治疗靶点。