Novel Analytical Platform For Robust Identification of Cell Migration Inhibitors
Novel Analytical Platform For Robust Identification of Cell Migration Inhibitors
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DOI：
10.1038/s41598-020-57806-0
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发表时间：
2020-01-22
影响因子：
4.6
通讯作者：
Sampattavanich, Somponnat
中科院分区：
文献类型：
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作者：
Somchai, Parinyachat;Phongkitkarun, Kriengkrai;Sampattavanich, Somponnat
伤口愈合试验是一种简单且成本效益高的体外试验，用于评估治疗对细胞迁移的影响。它的主要限制是可能与其他细胞表型混淆，导致对实验结果的误解。在这项研究中，我们试图通过开发一种简单的分析方法来评估治疗对细胞迁移和细胞死亡的影响，同时使用丝裂霉素C预处理使细胞生长的影响正常化，从而解决这个问题。通过仔细绘制细胞死亡和伤口愈合率之间的关系，可以在所有药物剂量下定量分离细胞死亡和细胞迁移对观察到的伤口愈合延迟的贡献。我们发现，内在的细胞运动差异和外在因素，如细胞播种密度，都能显著影响治疗对细胞表型影响的最终解释。这种差异可以通过使用每个治疗条件的实际伤口闭合时间来计算表型评分来纠正。最后，我们展示了对胆管癌细胞系中细胞迁移的强药物抑制剂的筛选。我们的方法能够准确地评分迁移和细胞毒性作用，并且可以很容易地用于高通量药物筛选。
Wound healing assay is a simple and cost-effective in vitro assay for assessing therapeutic impacts on cell migration. Its key limitation is the possible confoundment by other cellular phenotypes, causing misinterpretation of the experimental outcome. In this study, we attempted to address this problem by developing a simple analytical approach for scoring therapeutic influences on both cell migration and cell death, while normalizing the influence of cell growth using Mitomycin C pre-treatment. By carefully mapping the relationship between cell death and wound closure rate, contribution of cell death and cell migration on the observed wound closure delay can be quantitatively separated at all drug dosing. We showed that both intrinsic cell motility difference and extrinsic factors such as cell seeding density can significantly affect final interpretation of therapeutic impacts on cellular phenotypes. Such discrepancy can be rectified by using the actual wound closure time of each treatment condition for the calculation of phenotypic scores. Finally, we demonstrated a screen for strong pharmaceutical inhibitors of cell migration in cholangiocarcinoma cell lines. Our approach enables accurate scoring of both migrastatic and cytotoxic effects, and can be easily implemented for high-throughput drug screening.