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基于微流控芯片的非小细胞肺癌分期和转移中循环肿瘤细胞和干细胞的检测及诊断意义。

Detection of CTCs and CSCs in the staging and metastasis of non-small cell lung cancer based on microfluidic chip and the diagnostic significance.

机构信息

School of Pathology, Qiqihar Medical University, Qiqihar, Heilongjiang, P.R. China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Sep;24(18):9487-9496. doi: 10.26355/eurrev_202009_23034.

Abstract

OBJECTIVE

Dynamic monitoring of CTCs/CSCs can assist in the diagnosis and prognosis of tumors. This study explores the diagnostic significance of microfluidic chip technology in the detection of CTCs/CSCs in clinical staging and metastasis of patients with non-small cell lung cancer (NSCLC). That lays a solid foundation for the use of microfluidic chips to monitor CTCs/CSCs for the stage and metastasis of patients with non-small cell lung cancer.

PATIENTS AND METHODS

This study collected 80 patients with lung cancer from October 2017 to October 2018. Meanwhile, 30 healthy people and 30 patients with benign lung diseases were selected during the same period as the control group 1 and the control group 2, respectively. CellSearch (Huntington Valley, PA, USA) and microfluidic chip were used to detect CTCs, the sensitivities were recorded. ELISA methods were used to detect the concentrations of tumor markers VEGF-C, CEA, and CA125 in serum, and their association with CTCs and CSCs was analyzed. In addition, after 3 months, we followed up 40 patients with lung cancer, recorded their prognosis, and extracted peripheral blood to detect changes in their CTCs and CSCs. The CellSearch (Huntington Valley, PA, USA) system and the microfluidic chip system were used to detect the CTCs in patients with lung cancer, and the sensitivity and specificity of the patients were analyzed. The changes in CTCs and CSCs in the peripheral blood of the patient were recorded.

RESULTS

It can be seen that the positive rate of CTCs and CSCs is not significantly correlated with the patients' age, gender, pathological type (adenocarcinoma, squamous cell carcinoma), etc. They are significantly correlated with clinical stage (I + II and III + IV) and metastasis (metastasis and non-metastasis) (p<0.01). Then, we divided the patients into groups for testing, and analyzed the association between different groups of patients and CTCs and CSCs. Compared with control group 1 and control group 2, the positive rates of CTCs and CSCs in lung cancer metastasis group and non-metastasis group were significantly different (p<0.05). Compared with the control group 1 and control group 2, the positive rates of CTCs and CSCs in stage I + II and III + IV of lung cancer were significantly different (p<0.05). The positive rate was significantly higher in the cancer metastasis group (p<0.05). The concentrations of tumor markers VEGF-C, CEA, CA125 in the serum of patients were consistent with CTCs-negative and CTC-positive lung cancer, with significant differences (p<0.05). CSCs negative and CSCs positive patients have similar results. Subsequently, we analyzed the sensitivity and specificity of CSCs, CTCs, and tumor markers for the diagnosis of NSCLC. The results showed that the sensitivity of CSCs and CTCs to diagnose patients was significantly higher than that of tumor markers.

CONCLUSIONS

This study shows that our microfluidic chip device can exhibit relatively good performance and can better detect CTCs and CSCs. Monitoring CTCs and CSCs of patients can provide a basis for judging the stage and metastasis of patients.

摘要

目的

动态监测 CTCs/CSCs 有助于肿瘤的诊断和预后。本研究探讨了微流控芯片技术在检测非小细胞肺癌(NSCLC)患者临床分期和转移中 CTCs/CSCs 的诊断意义。为利用微流控芯片监测非小细胞肺癌患者的分期和转移奠定了坚实的基础。

患者和方法

本研究收集了 2017 年 10 月至 2018 年 10 月期间的 80 例肺癌患者。同时,选择同期的 30 名健康人和 30 名良性肺部疾病患者作为对照组 1 和对照组 2。使用 CellSearch(宾夕法尼亚州亨廷顿谷)和微流控芯片检测 CTCs,记录灵敏度。采用 ELISA 法检测血清中血管内皮生长因子-C(VEGF-C)、癌胚抗原(CEA)和 CA125 肿瘤标志物的浓度,并分析其与 CTCs 和 CSCs 的关系。此外,在 3 个月后,我们对 40 例肺癌患者进行了随访,记录了他们的预后,并提取外周血检测其 CTCs 和 CSCs 的变化。采用 CellSearch(宾夕法尼亚州亨廷顿谷)系统和微流控芯片系统检测肺癌患者的 CTCs,分析患者的灵敏度和特异性。记录患者外周血中 CTCs 和 CSCs 的变化。

结果

可见,CTCs 和 CSCs 的阳性率与患者的年龄、性别、病理类型(腺癌、鳞状细胞癌)等无明显相关性,与临床分期(I+II 和 III+IV)和转移(转移和非转移)明显相关(p<0.01)。然后,我们将患者分组进行检测,并分析了不同组患者与 CTCs 和 CSCs 的关系。与对照组 1 和对照组 2 相比,肺癌转移组和非转移组 CTCs 和 CSCs 的阳性率差异有统计学意义(p<0.05)。与对照组 1 和对照组 2 相比,I+II 期和 III+IV 期肺癌患者的 CTCs 和 CSCs 阳性率差异有统计学意义(p<0.05)。癌症转移组的阳性率明显较高(p<0.05)。血清中肿瘤标志物 VEGF-C、CEA、CA125 的浓度与 CTCs 阴性和 CTCs 阳性肺癌一致,差异有统计学意义(p<0.05)。CSCs 阴性和 CSCs 阳性患者的结果相似。随后,我们分析了 CSCs、CTCs 和肿瘤标志物对 NSCLC 诊断的灵敏度和特异性。结果表明,CSCs 和 CTCs 对患者的诊断灵敏度明显高于肿瘤标志物。

结论

本研究表明,我们的微流控芯片装置能够表现出较好的性能,能够更好地检测 CTCs 和 CSCs。监测患者的 CTCs 和 CSCs 可为判断患者的分期和转移提供依据。

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