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一种用于研究癌症转移中失巢凋亡抗性的实验工作流程。

An experimental workflow for investigating anoikis resistance in cancer metastasis.

作者信息

Han Xue, Zheng Yipan, Si Xiaohui, Liu Zhe

机构信息

Zhejiang Key Laboratory of Medical Epigenetics, School of Medicine, Hangzhou Normal University, Hangzhou 311121, China.

出版信息

J Biol Methods. 2025 Aug 5;12(3):e99010066. doi: 10.14440/jbm.2024.0140. eCollection 2025.

Abstract

BACKGROUND

Anoikis is a form of programmed cell death triggered by the detachment of cells from the extracellular matrix. Anoikis resistance represents a critical factor in tumor metastasis, and elucidating the mechanisms by which epithelial cancer cells evade this process may provide a molecular insight for effectively targeting metastatic progression.

METHODS

Presented here are an experimental workflow and a detailed protocol to examine anoikis sensitivity in tumor cells both and . We described a detachment-induced anoikis model, a three-dimensional spheroid culture system, and an circulating tumor cell assay, by using the human lung carcinoma cell line A549 as a model system. We detailed the cell culture conditions, materials, and sample preparation, and the evaluation and quantification of anoikis. Together, these methods provide a comprehensive approach for investigating anoikis resistance.

CONCLUSION

This protocol offers valuable insights into the mechanisms underlying anoikis resistance and may facilitate the identification of novel therapeutic targets for cancer treatment.

摘要

背景

失巢凋亡是一种由细胞与细胞外基质脱离所触发的程序性细胞死亡形式。失巢凋亡抗性是肿瘤转移的关键因素,阐明上皮癌细胞逃避这一过程的机制可能为有效靶向转移进展提供分子见解。

方法

本文介绍了一种用于检测肿瘤细胞失巢凋亡敏感性的实验工作流程和详细方案。我们以人肺癌细胞系A549作为模型系统,描述了一种脱离诱导的失巢凋亡模型、一种三维球体培养系统和一种循环肿瘤细胞检测方法。我们详细说明了细胞培养条件、材料、样品制备以及失巢凋亡的评估和定量。这些方法共同提供了一种研究失巢凋亡抗性的综合方法。

结论

该方案为失巢凋亡抗性的潜在机制提供了有价值的见解,并可能有助于识别癌症治疗的新治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1466/12422110/92dff8e47068/jbm-12-e99010066-g003.jpg

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