Exosomal miRNA Analysis in Non-small Cell Lung Cancer (NSCLC) Patients' Plasma Through qPCR: A Feasible Liquid Biopsy Tool

Marco Giallombardo; Jorge Chac&#225;rtegui Borr&#225;s; Marta Castiglia; Nele Van Der Steen; Inge Mertens; Patrick Pauwels; Marc Peeters; Christian Rolfo

doi:10.3791/53900

外来体miRNA的分析非小细胞肺癌（NSCLC）患者的血浆通过定量PCR:一种可行的液体活检工具

JoVE Journal
Medicine

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Medicine

Exosomal miRNA Analysis in Non-small Cell Lung Cancer (NSCLC) Patients’ Plasma Through qPCR: A Feasible Liquid Biopsy Tool

外来体miRNA的分析非小细胞肺癌（NSCLC）患者的血浆通过定量PCR:一种可行的液体活检工具

Full Text

11,673 Views

08:49 min

May 27, 2016

DOI: 10.3791/53900-v

Marco Giallombardo*^1,2, Jorge Chacártegui Borrás*^2,3, Marta Castiglia⁴, Nele Van Der Steen³, Inge Mertens^5,6, Patrick Pauwels^7,3, Marc Peeters^8,3, Christian Rolfo^2,3

¹Department of Biopathology and Medical Biotechnology, Section of Biology and Genetics,University of Palermo, ²Phase I-Early Clinical Trials Unit, Oncology Department,Antwerp University Hospital (UZA), ³Center for Oncological Research (CORE),Antwerp University, ⁴Department of Surgical, Oncological and Oral Sciences, Section of Medical Oncology,University of Palermo, ⁵Flemish Institute for Technological Research (VITO), ⁶CORE, Campus Groenenborger,Antwerp University, ⁷Molecular Pathology, Pathology Department,Antwerp University Hospital (UZA), ⁸Oncology Department,Antwerp University Hospital (UZA)

Please note that some of the translations on this page are AI generated. Click here for the English version.

该方案描述了通过具有蛋白酶 K 和 RNAse 处理的商业外泌体分离试剂盒在 NSCLC 患者血浆中释放的外泌体中进行 miRNA 分析的可行性，以避免循环 miRNA 污染并评估它们在 NSCLC 中的生物标志物特征。

该程序的总体目标是在外泌体分离之前使用双酶处理分离 NSCLC 患者血浆中存在的不含非外泌体微 RNA 来源（如自由循环微 RNAse）的外泌体。该方法可以帮助分析外泌体区室的微 RNA 脂质活检，这将有助于阐明它们作为生物标志物的用途。该程序的主要优点是酶促预处理，它消除了可能干扰分析的非外泌体 micro RNA 来源。

首先在冰上解冻 1 mL 血浆样品。解冻后，将试管倒置数次，以解聚可能形成的任何冷沉淀物。然后在室温下以 2， 000 G 离心血浆 20 分钟，以去除细胞和碎片。

View the full transcript and gain access to thousands of scientific videos