Viraka Nellore Bhanu Priya, Kanchanapally Rajashekhar, Pramanik Avijit, Sinha Sudarson Sekhar, Chavva Suhash Reddy, Hamme Ashton, Ray Paresh Chandra
Department of Chemistry and Biochemistry, Jackson State University , Jackson, Mississippi 39217, United States.
Bioconjug Chem. 2015 Feb 18;26(2):235-42. doi: 10.1021/bc500503e. Epub 2015 Jan 16.
Tumor metastasis is responsible for 1 in 4 deaths in the United States. Though it has been well-documented over past two decades that circulating tumor cells (CTCs) in blood can be used as a biomarker for metastatic cancer, there are enormous challenges in capturing and identifying CTCs with sufficient sensitivity and specificity. Because of the heterogeneous expression of CTC markers, it is now well understood that a single CTC marker is insufficient to capture all CTCs from the blood. Driven by the clear need, this study reports for the first time highly efficient capture and accurate identification of multiple types of CTCs from infected blood using aptamer-modified porous graphene oxide membranes. The results demonstrate that dye-modified S6, A9, and YJ-1 aptamers attached to 20-40 μm porous garphene oxide membranes are capable of capturing multiple types of tumor cells (SKBR3 breast cancer cells, LNCaP prostate cancer cells, and SW-948 colon cancer cells) selectively and simultaneously from infected blood. Our result shows that the capture efficiency of graphene oxide membranes is ~95% for multiple types of tumor cells; for each tumor concentration, 10 cells are present per milliliter of blood sample. The selectivity of our assay for capturing targeted tumor cells has been demonstrated using membranes without an antibody. Blood infected with different cells also has been used to demonstrate the targeted tumor cell capturing ability of aptamer-conjugated membranes. Our data also demonstrate that accurate analysis of multiple types of captured CTCs can be performed using multicolor fluorescence imaging. Aptamer-conjugated membranes reported here have good potential for the early diagnosis of diseases that are currently being detected by means of cell capture technologies.
在美国，肿瘤转移导致了四分之一的死亡。尽管在过去二十年里有充分的文献记载，血液中的循环肿瘤细胞（CTC）可作为转移性癌症的生物标志物，但在以足够的灵敏度和特异性捕获和识别CTC方面仍存在巨大挑战。由于CTC标志物的表达具有异质性，现在人们已经清楚地认识到单一的CTC标志物不足以从血液中捕获所有的CTC。出于明确的需求驱动，本研究首次报道了使用适体修饰的多孔氧化石墨烯膜从感染血液中高效捕获和准确识别多种类型的CTC。结果表明，附着在20 - 40μm多孔氧化石墨烯膜上的染料修饰的S6、A9和YJ - 1适体能够从感染血液中选择性地同时捕获多种类型的肿瘤细胞（SKBR3乳腺癌细胞、LNCaP前列腺癌细胞和SW - 948结肠癌细胞）。我们的结果表明，氧化石墨烯膜对多种类型肿瘤细胞的捕获效率约为95%；对于每种肿瘤浓度，每毫升血液样本中有10个细胞。我们使用不含抗体的膜证明了我们的检测方法捕获靶向肿瘤细胞的选择性。感染不同细胞的血液也被用于证明适体偶联膜捕获靶向肿瘤细胞的能力。我们的数据还表明，使用多色荧光成像可以对多种捕获的CTC进行准确分析。本文报道的适体偶联膜在目前通过细胞捕获技术检测的疾病早期诊断方面具有良好的潜力。
