LncRNA ILF3-AS1 Promotes the Progression of Colon Adenocarcinoma Cells Through the miR-619-5p/CAMK1D Axis.
LncRNA ILF3-AS1 Promotes the Progression of Colon Adenocarcinoma Cells Through the miR-619-5p/CAMK1D Axis.
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长链非编码RNA ILF3 - AS1通过miR - 619 - 5p / CAMK1D轴促进结肠腺癌细胞的进展。
DOI：
10.2147/ott.s296441
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发表时间：
2021
影响因子：
4
通讯作者：
Li S
中科院分区：
文献类型：
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作者：
Liu W;Li S
结肠腺癌（COAD）是消化道第三大常见肿瘤。近期研究报道，长链非编码RNA（lncRNA）的异常表达可能在结肠腺癌的发生和发展中起关键作用。
在本研究中，我们通过实时定量聚合酶链反应（RT - qPCR）研究了ILF3 - AS1在结肠腺癌细胞系、人正常结肠上皮细胞系、患者肿瘤组织及癌旁正常组织中的表达。将小干扰RNA（siRNAs）转染到结肠腺癌细胞中以抑制ILF3 - AS1的表达。分别通过CCK - 8实验、Transwell迁移和侵袭实验以及流式细胞术凋亡实验检测ILF3 - AS1对细胞增殖、迁移、侵袭和凋亡的影响。通过荧光素酶报告基因实验验证了ILF3 - AS1与miR - 619 - 5p / CAMK1D的直接结合。通过蛋白质印迹分析（Western Blot）检测CAMK1D和上皮 - 间质转化（EMT）相关蛋白的表达。此外，还进行了体内实验以证明ILF3 - AS1在结肠腺癌中的致癌作用。
结果显示，ILF3 - AS1在结肠腺癌组织中的表达显著高于癌旁正常样本，这一结论在现有的公共数据集中也得到了证实。在ILF3 - AS1敲低后，结肠腺癌细胞系SW480和HT29的增殖受到显著抑制。同时，细胞凋亡增加，侵袭和迁移能力下降。在进一步探究机制后，我们发现ILF3 - AS1作为miR - 619 - 5p的竞争性内源RNA，它可以与miR - 619 - 5p结合并降低其表达，从而调节靶基因CAMK1D。此外，我们发现ILF3 - AS1的高表达与结肠腺癌样本中的肿瘤分级、肿瘤大小和远处转移显著相关。体内实验证实ILF3 - AS1在结肠腺癌模型中促进肿瘤生长。
本研究表明，ILF3 - AS1通过调节miR - 619 - 5p / CAMK1D轴在结肠腺癌中发挥致癌作用，抑制ILF3 - AS1可能为结肠腺癌的治疗铺平道路。
Colon adenocarcinoma (COAD) is the third most common tumor of the digestive tract. Recent studies reported that lncRNA’s abnormal expression might play a vital role in the occurrence and development of COAD. In the present study, we investigated the expression of ILF3-AS1 in COAD cell lines, human normal colon epithelial cell line, patient tumor tissues and adjacent normal tissues by real-time quantitative PCR (RT-qPCR). Small interfering RNAs (siRNAs) were transfected into COAD cells to inhibit the expression of ILF3-AS1. The effects of ILF3-AS1 on cell proliferation, migration, invasion and apoptosis were measured by CCK-8 assay, transwell migration and invasion assay, and flow cytometry apoptosis assay, respectively. The direct binding of ILF3-AS1 and miR-619-5p/CAMK1D was validated by the luciferase reporter assay. The expression of CAMK1D and epithelial-mesenchymal transformation (EMT)-related proteins was detected by Western Blot analysis. Besides, in vivo experiments were conducted to demonstrate the oncogenic role of ILF3-AS1 in COAD. The results showed that the expression of ILF3-AS1 was significantly higher in COAD tissue than in normal adjacent samples, and this conclusion was confirmed in the available public datasets. After ILF3-AS1 knockdown, the proliferation of COAD cell lines SW480 and HT29 was significantly inhibited. At the same time, the apoptosis was increased, and the invasion and migration abilities were decreased. After further exploring the mechanisms, we found that ILF3-AS1 serves as a competitive endogenous RNA of mir-619-5p. It can bind to mir-619-5p and reduce its expression, thus regulating the target gene CAMK1D. In addition, we found that high expression of ILF3-AS1 was significantly associated with tumor grade, tumor size, and distant metastasis in COAD samples. In vivo experiments confirmed that ILF3-AS1 promotes tumor growth in COAD models. The present study demonstrated that ILF3-AS1 plays an oncogenic role in COAD through regulating the miR-619-5p/CAMK1D axis, and inhibition of ILF3-AS1 may pave the way for COAD treatment.