Background

The widely used in vitro invasion assays for head and neck squamous cell carcinoma (HNSCC) are wound healing, transwell, and organotypic assays. However, these are still lab-intensive and time-consuming tasks. For the rapid detection and high throughput screening of invasiveness in 3D condition, we propose a novel spheroid invasion assay using commercially available pillar platform system.

Materials and methods

Using the pillar-based spheroid invasion assay, migration and invasion was evaluated in three patient-derived cells (PDCs) of HNSCC. Immunofluorescence of live cells was used for the quantitative measurement of migratory and invaded cells attached to the pillar. Expression of epithelial-mesenchymal transition (EMT)-related gene (snai1/2) was measured by qRT-PCR. We also tested the impact of drug treatments (cisplatin, docetaxel) on the changes in the invasive phenotype.

Results

All PDCs successfully formed spheroid at 4 days and can be measured invasiveness within 7 days. Intriguingly, one PDC (#1) obtained from the advanced stage showed robust migration, invasion and higher transcription of snai1/2, compared with the other two PDCs. Furthermore, the invasion ratio of the control spheroids was about 70% while the invasion ratios of drug-treated spheroids were lower than 50%, and the difference showed statistical significance (p < 0.01).

Conclusion

The presented spheroid invasion assay using pillar array could be useful for the evaluation of cancer cell behavior and physiology in response to diverse therapeutic drugs.

中文翻译：

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头颈部鳞状细胞癌中侵袭性表型的快速定量：新型3D柱阵列系统。

背景

头颈部鳞状细胞癌（HNSCC）广泛使用的体外侵袭分析是伤口愈合，穿孔和器官型分析。但是，这些仍然是实验室密集且耗时的任务。为了在3D条件下快速检测侵袭性并进行高通量筛选，我们提出了使用可商购的柱平台系统进行新型球体侵袭试验。

材料和方法

使用基于支柱的球体入侵测定法，在HNSCC的三个患者来源细胞（PDC）中评估了迁移和侵袭。活细胞的免疫荧光用于定量测量附着在柱子上的迁移细胞和侵入细胞。通过qRT-PCR测量上皮-间质转化（EMT）相关基因（snai1 / 2）的表达。我们还测试了药物治疗（顺铂，多西他赛）对侵袭性表型变化的影响。

结果

所有PDC在第4天成功形成球体，并且可以在7天内测量侵袭性。有趣的是，与其他两个PDC相比，从晚期获得的一个PDC（＃1）显示出强劲的迁移，侵袭和snai1 / 2的更高转录。此外，对照球体的浸润率约为70％，而药物处理的球体的浸润率低于50％，差异具有统计学意义（p  <0.01）。

结论

提出的使用柱阵列的球体侵入测定法可用于评估响应各种治疗药物的癌细胞行为和生理学。