检测、分离和培养前列腺循环肿瘤细胞的新方法。

Novel method to detect, isolate, and culture prostate culturing circulating tumor cells.

作者信息

Ruan Diana, So Shuiping, King Bridgette, Wang Run

机构信息

Columbia University Vagelos College of Physician and Surgeons, New York, NY, USA.

Division of Urology, Department of Surgery, University of Texas Health Science Center-McGovern Medical School at Houston, University of Texas MD Anderson Cancer Center, Houston, TX, USA.

出版信息

Transl Androl Urol. 2019 Dec;8(6):686-695. doi: 10.21037/tau.2019.11.10.

DOI:10.21037/tau.2019.11.10

PMID:32038965

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6987604/

Abstract

BACKGROUND

Effectively detecting and culturing circulating tumor cells (CTCs), is critical for diagnosis of early metastasis, monitoring anti-cancer therapeutic efficacy, and drug screening. However, most current FDA approved CTC detection methods are based on antibody binding, which has limitations due to the nature of variations in antibody preparation and antibody-CTC size mismatches. Thus, searching for alternative and advanced methods is urgent and necessary.

METHODS

Prostate cancer tissue was digested by collagenase and cultured. Cancer stromal cells were identified and labelled with 4',6-diamidino-2-phenylindole (DAPI) before incubation with whole blood of cancer mice (bearing a later stage of prostate cancer). The attached blood CTCs on the DAPI-labeled cancer stromal cells were detected, isolated, cultured and produced into individual cancer cell lines.

RESULTS

Five clones of prostate cancer cells isolated from cancer tissue were successfully cultured. One (Clone-1) of the five clones showed positive staining for all three cancer stromal cell markers (CD133, α2β1 integrin and CD44). Clone-1 cells rich with epithelial cell adhesion molecule (EpCAM) on the cell surface were further identified. The Clone-1 stromal cells labeled as "bait" attracted and caught a trace number of CTCs from the whole blood of mice with advanced stage cancer. Efficient culturing of the caught CTCs from single cell to forming of individual cancer cell line(s) were established.

CONCLUSIONS

We present a fundamental advancement of CTC detection and culturing using a different mechanism (cell-cell interaction) rather than the traditional antibody-based immune-binding, such as CellSearch system. This study has potential to be fully developed into a novel approach for early cancer metastasis detection, and chemotherapy efficacy monitoring. The efficiently cultured CTCs could be used for single-clone CTC analysis and anti-cancer drug screening to further advance the development of individualized medicine.

摘要

背景

有效检测和培养循环肿瘤细胞（CTC）对于早期转移的诊断、监测抗癌治疗效果以及药物筛选至关重要。然而，目前大多数获得美国食品药品监督管理局（FDA）批准的CTC检测方法都基于抗体结合，由于抗体制备的差异以及抗体与CTC大小不匹配的性质，这种方法存在局限性。因此，寻找替代的先进方法迫在眉睫且十分必要。

方法

用胶原酶消化前列腺癌组织并进行培养。在与患有晚期前列腺癌的小鼠全血孵育之前，鉴定癌基质细胞并用4',6-二脒基-2-苯基吲哚（DAPI）进行标记。检测、分离、培养并将附着在DAPI标记的癌基质细胞上的血液CTC制成单个癌细胞系。

结果

成功培养了从癌组织中分离出的五个前列腺癌细胞克隆。这五个克隆中的一个（克隆-1）对所有三种癌基质细胞标志物（CD133、α2β1整合素和CD44）均呈阳性染色。进一步鉴定了细胞表面富含上皮细胞粘附分子（EpCAM）的克隆-1细胞。标记为“诱饵”的克隆-1基质细胞从晚期癌症小鼠的全血中吸引并捕获了少量CTC。建立了从单个细胞高效培养捕获的CTC直至形成单个癌细胞系的方法。