环状TBL1XR1/miR-424轴通过调控Smad7促进结直肠癌的增殖和转移。
CircTBL1XR1/miR-424 axis regulates Smad7 to promote the proliferation and metastasis of colorectal cancer.
作者信息
Li Na
机构信息
Department of Clinical Laboratory, First Hospital of Xi'an Jiaotong University, Xi'an, China.
出版信息
J Gastrointest Oncol. 2020 Oct;11(5):918-931. doi: 10.21037/jgo-20-395.
BACKGROUND
This study analyzed the effect of circular RNA (circRNA), TBL1XR1, on the malignant progression of colon cancer cells and explored its possible molecular mechanism.
METHODS
The expression levels of circTBL1XR1 in colorectal cancer and paracancerous tissues were detected by quantitative real-time polymerase chain (qRT-PCR). Changes in circTBL1XR1 expression in normal intestinal epithelial cells and colon cancer cell lines were detected at the cellular level. LoVo and SW620 cells with the highest circTBL1XR1 expression were transfected with circTBL1XR1, and the transfection efficiency was verified by qRT-PCR. The effects of circTBL1XR1 on the proliferation, migration, and invasion of colon cancer cells were detected by MTT, Transwell migration, and invasion assay, and a subcutaneous tumor model. Target genes of circTBL1XR1 were verified by luciferase reporter assay. Expression levels of circTBL1XR1 target genes were detected by qRT-PCR and Western blotting.
RESULTS
circTBL1XR1 was highly expressed in colon cancer patients. circTBL1XR1 expression in colon cancer cells was also higher than that in normal intestinal cells. and experimental results showed that overexpression of circTBL1XR1 enhanced the proliferation, migration, and invasion of colon cancer cells. After lowering circTBL1XR1, the ability of migration and invasion of colon cancer cells was significantly reduced. Bioinformatics retrieval and luciferase reporter gene assay confirmed that circTBL1XR1 can bind to microRNA-424 (miR-424) and that the Smad7 gene is the target gene of miR-424.
CONCLUSIONS
circTBL1XR1 was highly expressed in colon cancer, and miR-424 was poorly expressed in colon cancer cells. circTBL1XR1 regulates the expression of Smad7 through miR-424, thereby affecting the malignant progression of colorectal cancer.
背景
本研究分析了环状RNA（circRNA）TBL1XR1对结肠癌细胞恶性进展的影响，并探讨其可能的分子机制。
方法
采用定量实时聚合酶链反应（qRT-PCR）检测结直肠癌组织及癌旁组织中circTBL1XR1的表达水平。在细胞水平检测正常肠上皮细胞和结肠癌细胞系中circTBL1XR1表达的变化。对circTBL1XR1表达最高的LoVo和SW620细胞进行circTBL1XR1转染，并通过qRT-PCR验证转染效率。通过MTT法、Transwell迁移和侵袭实验以及皮下肿瘤模型检测circTBL1XR1对结肠癌细胞增殖、迁移和侵袭的影响。通过荧光素酶报告基因实验验证circTBL1XR1的靶基因。采用qRT-PCR和蛋白质免疫印迹法检测circTBL1XR1靶基因的表达水平。
结果
circTBL1XR1在结肠癌患者中高表达。结肠癌细胞中circTBL1XR1的表达也高于正常肠细胞。 实验结果表明，circTBL1XR1的过表达增强了结肠癌细胞的增殖、迁移和侵袭能力。降低circTBL1XR1后，结肠癌细胞的迁移和侵袭能力显著降低。生物信息学检索和荧光素酶报告基因实验证实，circTBL1XR1可与微小RNA-424（miR-424）结合，且Smad7基因是miR-424的靶基因。
结论
circTBL1XR1在结肠癌中高表达，miR-424在结肠癌细胞中低表达。circTBL1XR1通过miR-424调节Smad7的表达，从而影响结直肠癌的恶性进展。
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