National Facility for Protein Science in Shanghai, Shanghai Advanced Research Institute, Chinese Academy of Sciences, No. 99, Haike Road, Zhangjiang Hi-Tech Park, Pudong, Shanghai 201210, PR China.
National Facility for Protein Science in Shanghai, Shanghai Advanced Research Institute, Chinese Academy of Sciences, No. 99, Haike Road, Zhangjiang Hi-Tech Park, Pudong, Shanghai 201210, PR China.
Spectrochim Acta A Mol Biomol Spectrosc. 2022 Dec 15;283:121773. doi: 10.1016/j.saa.2022.121773. Epub 2022 Aug 18.
Doxorubicin is an effective chemotherapeutic agent applied in a wide variety of cancers. Despite its potent anticancer activity towards cancer cells, doxorubicin is also toxic to noncancerous cells. Therefore, doxorubicin can cause serious side effects in various organs, especially when dose escalation is required for patients with advanced disease. The liver is the major detoxification organ that metabolizes drugs, and hepatotoxicity is one of the most common adverse effects of doxorubicin administration. However, the exact mechanisms of doxorubicin-induced hepatotoxicity have not been clearly identified, and how doxorubicin treatment affects the biomolecular contents of normal human hepatocytes has rarely been studied. Synchrotron radiation-based Fourier transform infrared (SR-FTIR) microspectroscopy is a state-of-the-art analytical technique for characterizing the biomolecules present in cells. In this research, the biomolecular alterations of doxorubicin-treated normal human hepatocytes compared to untreated control cells were investigated at the single-cell level by combining SR-FTIR microspectroscopy with the Cell Counting Kit-8 (CCK-8) assay and flow cytometry. WRL68 human normal embryonic liver cells, which have been shown to be very promising for assessing the cytotoxicity of toxic compounds and investigating hepato-toxicology, were used in this research. Principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used to further analyse the biomolecular contents of WRL68 cells. The order of lipid acyl chains and protein α-helix structures in doxorubicin-treated WRL68 cells was found to be distinctly changed, while the nucleic acids were altered relatively less. No alteration in the carbohydrate content was distinguishable after doxorubicin treatment. These results provide more comprehensive information about the biomolecular changes in hepatocytes induced by doxorubicin treatment and help to elucidate the mechanism of doxorubicin-induced hepatotoxicity. This research also proves that SR-FTIR microspectroscopy, combined with PCA and OPLS-DA, is a promising approach for investigating drug-cell interaction systems.
多柔比星是一种有效的化疗药物，适用于多种癌症。尽管多柔比星对癌细胞具有强大的抗癌活性，但它对非癌细胞也有毒性。因此，多柔比星会在各种器官中引起严重的副作用，尤其是在需要对晚期疾病患者进行剂量升级时。肝脏是主要的解毒器官，可代谢药物，肝毒性是多柔比星给药的最常见不良反应之一。然而，多柔比星诱导的肝毒性的确切机制尚未明确，多柔比星治疗如何影响正常人类肝细胞的生物分子含量也很少被研究。基于同步辐射的傅里叶变换红外（SR-FTIR）微光谱学是一种用于表征细胞中存在的生物分子的最先进分析技术。在这项研究中，通过将 SR-FTIR 微光谱学与细胞计数试剂盒-8（CCK-8）测定法和流式细胞术相结合，在单细胞水平上研究了与未处理对照细胞相比，多柔比星处理的正常人类肝细胞的生物分子变化。WRL68 人正常胚胎肝细胞已被证明非常适合评估有毒化合物的细胞毒性并研究肝毒理学，因此本研究中使用了这些细胞。主成分分析（PCA）和正交偏最小二乘判别分析（OPLS-DA）用于进一步分析 WRL68 细胞的生物分子含量。发现多柔比星处理的 WRL68 细胞中脂质酰基链和蛋白质α-螺旋结构的顺序明显改变，而核酸的改变相对较少。多柔比星处理后，可区分的碳水化合物含量没有改变。这些结果提供了有关多柔比星处理诱导的肝细胞中生物分子变化的更全面信息，并有助于阐明多柔比星诱导的肝毒性机制。本研究还证明，SR-FTIR 微光谱学结合 PCA 和 OPLS-DA 是研究药物-细胞相互作用系统的有前途的方法。
