痰液中肺癌细胞分选及多种miRNA单细胞分析用于肺癌早期检测及分型

Lung cancer is the most common cause of cancer related deaths worldwide. Because lung cancer patients often have no obvious clinical symptoms in early stage of lung cancer and are easily missed, most of the cases are already diagnosed late (Phase III, IV). The severity of the disease is emphasized by the fact that the 5-year survival is only about 10%, one of the lowest among cancer patients. Therefore, early detection and precise diagnosis are critical for the patients to receive proper therapeutic treatment as early as possible. However, existing lung cancer detection methods still have more limitations on early detection of lung cancer and lung cancer screening of high-risk groups of lung cancer. Therefore, early detection of lung cancer is still challenging. Based on this, we use the characteristics of respiratory disease detection, innovatively proposed the use of microfluidic chip efficient isolation of lung cancer cells from sputum. Subsequently, a variety of miRNAs have been used as markers for the diagnosis and typing of lung cancers. Single-cell analysis of multiple miRNAs (> 4 species) have been developed for the mapping of multiple miRNA expression profiles in lung cancer cells for lung cancer early detection and histological typing. The project takes advantage of the advantages of early, non-invasive, simple and repeatable operation of sputum cytology to overcome its shortcomings of low sensitivity, difficult identification, poor biosecurity and unable to detect automated samples, to provide a new strategy and a new way for early non-invasive detection of lung cancer and practical screening of lung cancer in high-risk groups.

肺癌发病率及死亡率在恶性肿瘤中居于高位。通常情况下肺癌发病隐匿，早期常无明显的临床症状，易被漏诊，大多数病例确诊时已是晚期（III期、IV期），生存率很低。现有的肺癌检测手段在肺癌的早期检测上和肺癌高危人群筛查上仍有较多限制，因此，实现肺癌早期检测依然充满挑战。由此，我们基于肺癌为呼吸系统疾病这一特点，创新性地提出应用微流控芯片高效分选痰液中的肺癌细胞，以多种miRNA作为肺癌诊断和分型的标志物分子，发展多种miRNA（>4种）同时检测的单细胞分析方法，描绘肺癌细胞内多种miRNA表达谱，进行肺癌的早期检测与组织学分型。该项目发挥传统痰液细胞学检查早期、无创伤、简便易行及可重复操作等优势，克服其临床检查方法的敏感度低、识别困难、生物安全性差与无法实现自动化样品检测等缺点，为肺癌的早期无创检测与高危人群实用性肺癌筛选提供新的策略与新的途径。

项目以肺癌患者痰液样品中脱落肿瘤细胞为分析对象，构建了集成分选和分析功能的微流控系统用于痰液中肿瘤细胞富集、分选及相关肿瘤功能标志物单细胞分析。该方法实现了临床肺鳞癌和肺腺癌患者痰液样品中脱落肿瘤细胞的区分和确认，检测结果与临床诊断结果具有高度相关性。进一步地，相似的方法也被应用到血液中循环肿瘤细胞的分选、表型分析及抗药能力等研究中。研究成果有望为肺癌患者的非侵入性检测和高危人群实用性肺癌筛查提供一种新的策略。相关研究成果已经在 Chem. Sci.， Anal. Chem.， Chem. Commun. 等杂志上发表学术论文4 篇，申请国家发明专利1 项。项目负责人参加研发的肿瘤细胞分选和原位荧光成像相关设备正进行仪器研制和临床合作。项目执行过程共协助培养硕士研究生2 名，项目负责人获评副教授。