Department of General Surgery, First Affiliated Hospital of China Medical University, Shenyang Liaoning, P.R. China.
Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Liver Disease Research Institute of Shanghai Institute of Traditional Chinese Medicine, Key Laboratory of Liver and Kidney Diseases of the Ministry of Education, Shanghai, China.
Cell Cycle. 2021 Dec;20(23):2476-2493. doi: 10.1080/15384101.2021.1988227. Epub 2021 Oct 31.
CircRNA/miRNA/mRNA axis has been reported to play crucial regulatory roles in multiple cancers, including hepatocellular carcinoma (HCC). In addition, recent investigations revealed that aloin exerted anti-tumor functions in HCC. However, the underlying mechanism of aloin on anti-tumor functions in HCC remained elusive. Therefore, this study aimed to investigate whether circRNA/miRNA/mRNA axis medicated the anti-tumor effect of aloin in HCC. Cell viability, invasion, apoptosis and autophagy were accessed by cell counting kit-8 (CCK-8), transwell invasion assay, flow cytometry, Western blot and immunofluorescence analysis, respectively. Expression levels of circ_0011385, miR-149-5p and WT1 mRNA were determined using qRT-PCR assay. Binding sites between miR-149-5p and circ_0011385 or WT1 were predicted in starBase database. The binding relationship among circ_0011385, miR-149-5p and WT1 were verified by dual-luciferase reporter assay and RNA immunoprecipitation. Besides, the rescue experiments were performed by co-transfection with cric_0011385 overexpression plasmid, si-cric_0011385, miR-149-5p mimic and inhibitor, WT1 pDNA and si-WT1 in HCC cells. Furthermore, tumor growth was also investigated in the xenograft mouse model. Aloin inhibited HCC proliferation and invasion as well as promoted apoptosis and autophagy both and . Besides, aloin suppressed circ_0011385 expression. Overexpressed circ_0011385 partially reversed the anti-tumor effect of aloin on HCC. In addition, it was revealed that the circ_0011385, miR-149-5p and WT1 genes were abnormally expressed in HCC. Furthermore, the binding interactions between circ_0011385, miR-149-5p and WT1 were predicted and confirmed. Moreover, the effect of circ_0011385 on the anti-tumor role of aloin in HCC was rescued by miR-149-5p mimics. MiR-149-5p regulated HCC progression via modulating WT1. Aloin suppressed cell proliferation, invasion and tumor growth and promoted apoptosis and autophagy in HCC through regulating circ_0011385/miR-149-5p/WT1 axis. Aloin may be a potential candidate drug for HCC treatment. HCC: Hepatocellular carcinoma; ceRNA: competing endogenous RNA; miRNA: microRNA; MREs: miRNA response elements; WT1: Wilms' tumor 1; MMP-2: Matrix metalloproteinase; EMT: epithelial-mesenchymal transition; GADPH: glyceraldehyde 3-phosphate dehydrogenase; WT: wild type; MUT: mutant type; DMEM: dulbecco's modified eagle medium.
环状 RNA/miRNA/mRNA 轴已被报道在多种癌症中发挥关键的调节作用，包括肝细胞癌 (HCC)。此外，最近的研究表明，芦荟素在 HCC 中具有抗肿瘤作用。然而，芦荟素在 HCC 中的抗肿瘤作用的潜在机制仍不清楚。因此，本研究旨在探讨环状 RNA/miRNA/mRNA 轴是否介导了芦荟素在 HCC 中的抗肿瘤作用。通过细胞计数试剂盒-8 (CCK-8)、Transwell 侵袭实验、流式细胞术、Western blot 和免疫荧光分析分别检测细胞活力、侵袭、凋亡和自噬。采用 qRT-PCR 检测 circ_0011385、miR-149-5p 和 WT1 mRNA 的表达水平。在 starBase 数据库中预测 miR-149-5p 与 circ_0011385 或 WT1 的结合位点。通过双荧光素酶报告基因检测和 RNA 免疫沉淀验证 circ_0011385、miR-149-5p 和 WT1 之间的结合关系。此外，还通过共转染 circ_0011385 过表达质粒、si-circ_0011385、miR-149-5p 模拟物和抑制剂、WT1 pDNA 和 si-WT1 在 HCC 细胞中进行了挽救实验。此外，还在异种移植小鼠模型中研究了肿瘤生长情况。芦荟素抑制 HCC 增殖和侵袭，促进凋亡和自噬。此外，芦荟素抑制 circ_0011385 的表达。过表达 circ_0011385 部分逆转了芦荟素对 HCC 的抗肿瘤作用。此外，还发现 circ_0011385、miR-149-5p 和 WT1 基因在 HCC 中异常表达。此外，还预测并证实了 circ_0011385、miR-149-5p 和 WT1 之间的结合相互作用。此外，miR-149-5p 模拟物可以挽救 circ_0011385 对芦荟素抑制 HCC 中抗肿瘤作用的影响。miR-149-5p 通过调节 WT1 来调节 HCC 的进展。芦荟素通过调节 circ_0011385/miR-149-5p/WT1 轴抑制 HCC 细胞增殖、侵袭和肿瘤生长，促进凋亡和自噬。芦荟素可能是 HCC 治疗的潜在候选药物。HCC：肝细胞癌；ceRNA：竞争内源 RNA；miRNA：微小 RNA；MREs：miRNA 反应元件；WT1：Wilms 肿瘤 1；MMP-2：基质金属蛋白酶；EMT：上皮-间充质转化；GAPDH：甘油醛 3-磷酸脱氢酶；WT：野生型；MUT：突变型；DMEM：杜尔贝科改良伊格尔培养基。
