He Fei, Zhong Xuejing, Lin Zheng, Lin Jianbo, Qiu Minglian, Li Xu, Hu Zhijian
Department of Epidemiology and Health Statistics, School of Public Health, Fujian Medical University, Fuzhou, 350108, China; Fujian Provincial Key Laboratory of Tumor Microbiology, Fujian Medical University, Fuzhou, 350108, China; Key Laboratory of Ministry of Education for Gastrointestinal Cancer, Fujian Medical University, Fuzhou, 350108, China.
Department of Science and Education, The Affiliated Longyan First Hospital of Fujian Medical University, Longyan, 364000, China.
J Cancer. 2020 Apr 6;11(14):4037-4046. doi: 10.7150/jca.30360. eCollection 2020.
: To investigate the relationship between CXCR4-related circular RNAs (circRNAs) in exosomes and lymph node metastasis of lung adenocarcinoma. : Totally 41 lung adenocarcinoma tissues (21 with lymph node metastasis and 20 without) were collected. Expression of CXCR4 protein was detected by western blotting analysis. A stable PC9/CXCR4-shRNA and PC14/CXCR4-shRNA knockdown lung adenocarcinoma cell lines were established and subjected to functional assays (cell proliferation, colony formation, migration and invasion) for phenotype changes. Exo-hsa-circRNAs (has-circRNAs in exosomes) were detected and in . The diagnostic value of differentially expressed exo-has-circRNAs was evaluated. : Expression levels of CXCR4 were higher in patients with lymph node metastasis than in those without ( = 0.001). Silencing expression in PC9 and PC14 cell lines with short hairpin RNA could effectively abolish colony formation frequency, proliferation rate, migration rate, and the number of invasive cells (all < 0.001). Exo_circRNA_0056616 was detected in both PC-9/CXCR4-shRNA cells and lung adenocarcinoma plasma at significantly higher levels than in the corresponding control ( < 0.001). When a receiver operating characteristic (ROC) curve for plasma exo-hsa_circRNA_0056616 levels and diagnosis of lymph node metastasis of lung adenocarcinoma was generated, a cutoff value of 0.394 was identified with an area under the curve of 0.812 (95% confidence interval 0.720-0.903), a sensitivity of 0.792, and specificity of 0.810. : Taken together, our findings suggested that CXCR4 was higher in the lung adenocarcinoma tissues with lymph node metastasis. Higher plasma levels of exo-hsa_circRNA_0056616 in these patients also suggest that this circRNA represents a potential biomarker for lymph node metastasis predictor in lung adenocarcinoma.
探讨外泌体中CXCR4相关环状RNA（circRNA）与肺腺癌淋巴结转移的关系。方法：收集41例肺腺癌组织（21例有淋巴结转移，20例无淋巴结转移）。采用蛋白质免疫印迹分析检测CXCR4蛋白表达。构建稳定的PC9/CXCR4-shRNA和PC14/CXCR4-shRNA敲低肺腺癌细胞系，并进行功能实验（细胞增殖、集落形成、迁移和侵袭）以观察表型变化。检测外泌体中的外显子-人环状RNA（exo-hsa-circRNA，外泌体中的hsa-circRNA）。评估差异表达的外显子-人环状RNA的诊断价值。结果：有淋巴结转移患者的CXCR4表达水平高于无淋巴结转移患者（P = 0.001）。用短发夹RNA沉默PC9和PC14细胞系中的CXCR4表达可有效降低集落形成频率、增殖率、迁移率和侵袭细胞数量（均P < 0.001）。PC-9/CXCR4-shRNA细胞和肺腺癌血浆中外显子环状RNA_0056616的检测水平均显著高于相应对照（P < 0.001）。绘制血浆外显子-人环状RNA_0056616水平对肺腺癌淋巴结转移诊断的受试者工作特征（ROC）曲线，确定截断值为0.394，曲线下面积为0.812（95%置信区间0.720 - 0.903），灵敏度为0.792，特异性为0.810。结论：综上所述，我们的研究结果表明，有淋巴结转移的肺腺癌组织中CXCR4表达较高。这些患者血浆中外显子-人环状RNA_0056616水平较高也表明，这种环状RNA代表了肺腺癌淋巴结转移预测的潜在生物标志物。
