Zuo Li, Li Xiaoli, Zhu Hailong, Li Anqi, Wang Yonggang
Department of Oncology, Fudan University Shanghai Cancer Center Minhang Branch Hospital, Ruili Road, Shanghai 201100, China.
Department of Oncology, Affiliated Sixth People's Hospital of Shanghai Jiaotong University, Yishan Road, Shanghai 200030, China.
ACS Omega. 2021 Aug 19;6(34):22011-22019. doi: 10.1021/acsomega.1c02425. eCollection 2021 Aug 31.
To determine the possibility of early diagnosis and prognosis of ovarian cancer (OC) via detecting miR-181a in circulating tumor cells (CTCs) of OC and to solve clinical difficulties in OC tissue sample collection. EpCAM liposome magnetic beads (Ep-LMBs) were prepared by the reverse-phase evaporation method, and the performance of EpCAM was characterized. The cytotoxicity assay was detected by the MTT assay, and CTC capture efficiency was determined using OC cell lines. Blood and tissue samples were collected from 30 patients with OC and 30 normal ovarian tissue samples were selected. Expression of miR-181a in CTCs and tissue samples was measured by real-time fluorescence quantitative PCR (RT-qPCR) with U6 as an internal reference. Expression of miR-181a was interfered in OC cells and its relative expression was measured. Ep-LMBs were successfully prepared with high stability. Cellular assays showed that these Ep-LMBs could capture up to 80% of OC cells. RT-qPCR showed that the expression of miR-181a was increased in OC tissues compared with that in normal ovarian tissues, and the relative expressions of miR-181a in cancerous tissues and CTCs were comparable. Correlation analysis with clinical characteristics revealed that miR-181a expression was correlated with the stage and metastasis of OC and the difference was statistically significant. MiR-181a may be involved in the development and progression of OC as an oncogene. Detection of miR-181a in Ep-LMB-captured CTCs is an effective and feasible alternative method for early diagnosis and prognostic evaluation of OC other than tissue tests.
通过检测卵巢癌(OC)循环肿瘤细胞(CTC)中的miR-181a来确定OC早期诊断和预后的可能性,并解决OC组织样本采集的临床难题。采用反相蒸发法制备上皮细胞黏附分子脂质体磁珠(Ep-LMBs),并对EpCAM的性能进行表征。通过MTT法检测细胞毒性,使用OC细胞系测定CTC捕获效率。收集30例OC患者的血液和组织样本,并选取30例正常卵巢组织样本。以U6为内参,通过实时荧光定量PCR(RT-qPCR)检测CTC和组织样本中miR-181a 的表达。干扰OC细胞中miR-181a的表达并测定其相对表达量。成功制备了具有高稳定性的Ep-LMBs。细胞实验表明,这些Ep-LMBs可捕获高达80%的OC细胞。RT-qPCR显示,与正常卵巢组织相比,OC组织中miR-181a的表达增加,癌组织和CTC中miR-181a的相对表达相当。与临床特征的相关性分析显示,miR-181a表达与OC的分期和转移相关,差异具有统计学意义。miR-181a可能作为癌基因参与OC的发生发展。检测Ep-LMB捕获的CTC中的miR-181a是一种除组织检测外用于OC早期诊断和预后评估的有效且可行的替代方法。