Department of Thoracic Surgery, The First Affiliated Hospital of China Medical University, Shenyang, China.
Eur Rev Med Pharmacol Sci. 2020 Sep;24(18):9511-9521. doi: 10.26355/eurrev_202009_23036.
The purpose of this study was to investigate the correlations of hsa_circ_0046264 expression with the onset, pathological stage, and chemotherapy resistance of lung cancer.
Firstly, gene expression profiling microarrays were applied to screen the differentially expressed circular ribonucleic acids (circRNAs) in the tumor tissues of patients with non-small cell lung cancer (NSCLC). Secondly, quantitative Reverse Transcription-Polymerase Chain Reaction (RT-qPCR) assay was adopted to further verify the circRNAs with significant differences. Thirdly, the correlations of hsa_circ_0046264 expression level with the clinical features of NSCLC patients were explored via statistical analysis. Fourthly, Kaplan-Meier survival analysis and receiver operating characteristic (ROC) curve were utilized to investigate the influence of hsa_circ_0046264 expression level on the survival of the patients. Finally, the role of hsa_circ_0046264 in the process of lung cancer was probed using in vitro experimental methods.
It was shown in the results of gene microarray assay that hsa_circ_0046264 was the most prominently upregulated gene, and RT-qPCR assay further proved that hsa_circ_0046264 expression was upregulated remarkably in the tissues of tumor patients. Clinical analysis indicated that the expression level of hsa_circ_0046264 was notably associated with the patient's age, tumor size, tumor-node-metastasis (TNM) stage, and lymph node metastasis (p<0.01). In addition, Kaplan-Meier statistical analysis manifested that the patients in hsa_circ_0046264 low-expression group had a markedly longer survival than those in hsa_circ_0046264 high-expression group. In the tumor tissues and serum of the patients, the area under ROC curve of hsa_circ_0046264 was 0.971 and 0.915, the specificity was 0.973 and 0.957, and the sensitivity was 0.951 and 0.927, while the Youden Index was 0.924 and 0.884 respectively. The results of Cell Counting Kit-8 (CCK-8) assay revealed that the proliferative ability of lung cancer A549 cells was significantly enhanced at 36, 48, and 72 h in hsa_circ_0046264 overexpression group. According to the results of wound-healing assay, the migratory ability of A549 cells was distinctly strengthened in hsa_circ_0046264 overexpression group compared with that in the control group (p<0.05). Moreover, the transwell assay results pointed out that the invasive ability of A549 cell lines at 48 h after overexpression of hsa_circ_0046264 was evidently stronger than that in control group (p<0.05). Under the stimulation of different doses of cisplatin, hsa_circ_0046264 overexpression group had a clearly raised survival rate of A549 cells in comparison with control group, and the differences in data were statistically significant (p<0.01).
Hsa_circ_0046264 may serve as a potential biomarker for the diagnosis and prognosis and a possible therapeutic target of lung cancer.
本研究旨在探讨 hsa_circ_0046264 的表达与肺癌的发病、病理分期和化疗耐药性的相关性。
首先，应用基因表达谱微阵列筛选非小细胞肺癌（NSCLC）患者肿瘤组织中差异表达的环状 RNA（circRNAs）。其次，采用定量逆转录-聚合酶链反应（RT-qPCR）检测进一步验证差异显著的 circRNAs。第三，通过统计学分析探讨 hsa_circ_0046264 表达水平与 NSCLC 患者临床特征的相关性。第四，采用 Kaplan-Meier 生存分析和受试者工作特征（ROC）曲线探讨 hsa_circ_0046264 表达水平对患者生存的影响。最后，采用体外实验方法探讨 hsa_circ_0046264 在肺癌发生发展过程中的作用。
基因微阵列检测结果显示 hsa_circ_0046264 是最显著上调的基因，RT-qPCR 检测进一步证实 hsa_circ_0046264 在肿瘤患者组织中显著上调。临床分析表明，hsa_circ_0046264 的表达水平与患者的年龄、肿瘤大小、肿瘤-淋巴结-转移（TNM）分期和淋巴结转移显著相关（p<0.01）。此外，Kaplan-Meier 统计分析表明，hsa_circ_0046264 低表达组患者的生存时间明显长于 hsa_circ_0046264 高表达组。在患者的肿瘤组织和血清中，hsa_circ_0046264 的 ROC 曲线下面积分别为 0.971 和 0.915，特异性分别为 0.973 和 0.957，敏感性分别为 0.951 和 0.927，而 Youden 指数分别为 0.924 和 0.884。CCK-8 检测结果显示，hsa_circ_0046264 过表达组在 36、48 和 72 h 时肺癌 A549 细胞的增殖能力显著增强。根据划痕愈合实验的结果，hsa_circ_0046264 过表达组 A549 细胞的迁移能力明显强于对照组（p<0.05）。此外，transwell 实验结果表明，hsa_circ_0046264 过表达组在过表达 48 小时后 A549 细胞系的侵袭能力明显强于对照组（p<0.05）。在不同剂量顺铂的刺激下，hsa_circ_0046264 过表达组 A549 细胞的存活率明显高于对照组，数据差异具有统计学意义（p<0.01）。
hsa_circ_0046264 可能作为肺癌诊断和预后的潜在生物标志物和可能的治疗靶点。
