Department of Mechanical Engineering, Indian Institute of Technology, Madras, Chennai, India.
Department of Biotechnology, Indian Institute of Technology, Madras, Room No. 201, Biotech Old Building, Chennai, India.
Lab Chip. 2022 May 31;22(11):2200-2211. doi: 10.1039/d1lc01139a.
Fluid shear stress (FSS) is crucial in cancer cell survival and tumor development. Noteworthily, cancer cells are exposed to several degrees of FSS in the tumor microenvironment and during metastasis. Consequently, the stemness marker expression in cancer cells changes with the FSS signal, although it is unclear how it varies with different magnitudes and during metastasis. The current work explores the stemness and drug resistance characteristics of the cervical cancer cell line HeLa in a microfluidic device with a wide range of physiological FSS. Hence, the microfluidic device was designed to achieve a logarithmic flow distribution in four culture chambers, realizing four orders of biological shear stress on a single chip. The cell cycle analysis demonstrated altered cell proliferation and mitotic arrest after FSS treatment. In addition, EdU staining revealed increased cell proliferation with medium to low FSS, whereas high shear had a suppressing effect. FSS increased competence to withstand higher intracellular ROS and mitochondrial membrane potential in HeLa. Furthermore, stemness-related gene (Sox2, N-cadherin) and cell surface marker (CD44, CD33, CD117) expressions were enhanced by FSS mechanotransduction in a magnitude-dependent manner. In summary, these stemness-like properties were concurrent with the drug resistance capability of HeLa towards doxorubicin. Overall, our microfluidic device elucidates cancer cell survival and drug resistance mechanisms during metastasis and in cancer relapse patients.
流体切应力（FSS）对癌细胞的存活和肿瘤的发展至关重要。值得注意的是，癌细胞在肿瘤微环境中和转移过程中会受到几度的 FSS 作用。因此，尽管目前尚不清楚 FSS 信号如何随不同幅度和转移而变化，但癌细胞中的干性标志物表达会随 FSS 信号而改变。本研究在具有广泛生理 FSS 的微流控装置中探索了宫颈癌 HeLa 细胞系的干性和耐药性特征。因此，该微流控装置旨在实现四个培养室中的对数式流量分布，从而在单个芯片上实现四个数量级的生物切应力。细胞周期分析表明 FSS 处理后细胞增殖发生改变并出现有丝分裂阻滞。此外，EdU 染色显示中等至低 FSS 可促进细胞增殖，而高剪切力则具有抑制作用。FSS 增加了 HeLa 细胞承受更高的细胞内 ROS 和线粒体膜电位的能力。此外，FSS 机械转导以幅度依赖的方式增强了干性相关基因（Sox2、N-钙黏蛋白）和细胞表面标志物（CD44、CD33、CD117）的表达。总之，这些类干性特性与 HeLa 对阿霉素的耐药能力相关。总体而言，我们的微流控装置阐明了转移过程中和癌症复发患者中癌细胞的存活和耐药机制。
