Tang Ying, Gao Zhikui, Liu Ran
Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing, China 210009.
Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing, China 210009.
Pathol Res Pract. 2022 May;233:153875. doi: 10.1016/j.prp.2022.153875. Epub 2022 Apr 1.
Esophageal squamous cell carcinoma (ESCC) has a high degree of malignancy, and there is currently no effective treatment. Circular RNA is a novel endogenous noncoding RNA with a stable loop structure. Several theories have been proposed regarding its biogenesis and usefulness as a biomarker in various cancers. Here we explore the predictive value and potential functions of a candidate circRNA (hsa_circ_0071106) in ESCC.
The altered expression of hsa_circ_0071106 was validated in clinical tissue samples from 64 patients with ESCC. The correlation between the clinical characteristics of these patients and the expression of hsa_circ_0071106 was analyzed by the chi-square test. The receiver operating characteristic (ROC) curves and the area under the ROC curve (AUC) were used to evaluate the diagnostic value of hsa_circ_0071106. The human esophageal carcinoma cell lines ECA109 and ECA9706 and the normal human esophageal epithelial cell line (HET-1A) were applied for functional analysis of hsa_circ_0071106. qRT-PCR, sanger sequencing, agarose gel electrophoresis, RNase R digestion assay, Actinomycin D inhibition assay, nucleoplasm separation assay, and FISH assays were utilized to verify the circular characteristics and subcellular localization of hsa_circ_0071106. SiRNA and circRNA interference lentivirus was constructed to inhibit the expression of hsa_circ_0071106 in the ECA109 and ECA9706 cell lines. We used the EdU assay, Transwell assay, and flow cytometry to detect changes in cell proliferation, invasion, migration, and apoptosis.
We identified that hsa_circ_0071106 showed significantly decreased expression levels in ESCC cells (P < 0.001) and tissue samples (P < 0.001). The correlation analysis of clinicopathological features and gene expression revealed that low expression of hsa_circ_0071106 was related to poor differentiation grade (P < 0.05). ROC curves indicated that hsa_circ_0071106 might have a diagnostic value for ESCC (AUC = 0.826, 95% CI: 0.748-0.904) and a predictive value for a high degree of histological differentiation (AUC = 0.730, 95% CI: 0.607-0.854). Hsa_circ_0071106 has a stable circular structure and was primarily localized in the cytoplasm. A transient knockdown of hsa_circ_0071106 by siRNA promotes proliferation, migration, and invasion and inhibits apoptosis of ESCC cells. A steady knockdown of hsa_circ_0071106 by a lentivirus knockout vector significantly promoted cell migration and invasion in ECA109 and ECA9706 cell lines.
Hsa_circ_0071106 was a downregulated circRNA related to ESCC and promoted ESCC progression by regulating cell migration and invasion. These findings suggest that hsa_circ_0071106 could be a promising diagnostic biomarker and potential therapeutic target and might predict ESCC's histological differentiation degree.
食管鳞状细胞癌（ESCC）恶性程度高，目前尚无有效治疗方法。环状RNA是一种具有稳定环状结构的新型内源性非编码RNA。关于其生物发生及其作为各种癌症生物标志物的用途，已经提出了几种理论。在此，我们探讨候选环状RNA（hsa_circ_0071106）在ESCC中的预测价值和潜在功能。
在64例ESCC患者的临床组织样本中验证了hsa_circ_0071106的表达改变。通过卡方检验分析这些患者的临床特征与hsa_circ_0071106表达之间的相关性。采用受试者工作特征（ROC）曲线和ROC曲线下面积（AUC）评估hsa_circ_0071106的诊断价值。将人食管癌细胞系ECA109和ECA9706以及正常人食管上皮细胞系（HET-1A）用于hsa_circ_0071106的功能分析。采用qRT-PCR、桑格测序、琼脂糖凝胶电泳、RNase R消化试验、放线菌素D抑制试验、核质分离试验和FISH试验验证hsa_circ_0071106的环状特征和亚细胞定位。构建siRNA和环状RNA干扰慢病毒以抑制ECA109和ECA9706细胞系中hsa_circ_0071106的表达。我们使用EdU试验、Transwell试验和流式细胞术检测细胞增殖、侵袭、迁移和凋亡的变化。
我们发现hsa_circ_0071106在ESCC细胞（P < 0.001）和组织样本（P < 0.001）中的表达水平显著降低。临床病理特征与基因表达的相关性分析显示，hsa_circ_0071106低表达与低分化程度相关（P < 0.05）。ROC曲线表明，hsa_circ_0071106可能对ESCC具有诊断价值（AUC = 0.826，95%CI：0.748 - 0.904），对高组织学分化程度具有预测价值（AUC = 0.730，95%CI：0.607 - 0.854）。Hsa_circ_0071106具有稳定的环状结构，主要定位于细胞质中。通过siRNA瞬时敲低hsa_circ_0071106可促进ESCC细胞的增殖、迁移和侵袭并抑制其凋亡。通过慢病毒敲除载体稳定敲低hsa_circ_0071106可显著促进ECA109和ECA9706细胞系中的细胞迁移和侵袭。
Hsa_circ_0071106是一种与ESCC相关的下调环状RNA，通过调节细胞迁移和侵袭促进ESCC进展。这些发现表明，hsa_circ_0071106可能是一种有前景的诊断生物标志物和潜在治疗靶点，并可能预测ESCC的组织学分化程度。
