| Zinc oxide nanorods(ZnO NRs)have garnered significant attention in recent years due to their ability to selectively target and kill cancer cells.The mechanism behind this process has been preliminarily proven:the dissolution of zinc ions and excess generation of reactive oxygen species induces oxidative stress,which ultimately leads to cancer cell apoptosis.The active targeting of ZnO NRs to cancer cells minimizes damage to healthy cells and reduces nonspecific killing,making them a promising new material for anticancer treatment.However,previous studies have focused on the analysis of apoptotic genes and corresponding proteins in cells,overlooking the fact that cells are complex organisms with interconnected physiological activities and functions.Additionally,prior researches have primarily utilized population-cell analysis,which failed to capture the critical role of cell heterogeneity in tumor evolution and overlooked essential information regarding cancer treatment.Herein,this.thesis utilized microfluidic chips based on microelectronic technology and bio-sensing nanomaterials,graphene oxide quantum dots(GOQDs),to analyze functional cytokines secrected from cells.It investigates the inhibitory effect of ZnO nanorods on breast cancer cells at both the population and single-cell levels.The main research contents of this thesis are as follows:The inhibitory effect of ZnO NRs on breast cancer population cells was studied by microfluidic biochips.MDA-MB-231 and MCF-7 cells were severally cultured with hydrothermally prepared ZnO NRs,which were single-crystal hexagonal nanorods with a diameter of 152.6 ± 28.2 nm and a vertical height of approximately 1240 nm,as confirmed by multiple material characterization experiments.The microfluidic chips were fabricated using photolithography technology,and GOQDs sensing detection substrate were prepared using nanomaterial self-assembly technology,which were used to detect 12 cytokines in a highthroughput manner.ZnO NRs promoted the secretion of HSP70 factors from MDA-MB-231 and MCF-7 cells while inhibiting the secretion of other 11 cytokines,indicating that cells were prevented from proliferating and metastasizing.The effect of ZnO NRs on breast cancer cell heterogeneity and cluster functions was investigated through single-cell microchips.The microchips used for MDA-MB-231 and MCF7 single cells culture were prepared using photolithography technology.The capture antibody barcode substrates were utilized to collect cytokines secreted by individual cells.The t-SNE and community-based clustering methods were employed to analyze the resulting data and visualize the secretion of 12 cytokines,thus forming multiple cell clusters with different plienotypes and functions.Spearman analysis indicated that ZnO NRs altered the functions of IFN-γ.TNF-α.and Granzyme B.and weakened the correlation between elusters,suggesting a greater inhibitory effect on proliferative functional clusters than metastatic functional clusters.These findings implied that ZnO NRs could pose a risk to the treatment of highly invasive breast cancer cells,such as MDA-MB-231.This thesis provided,for the first time,a single-ceIl resolution analysis of the tumor suppressor effect of ZnO NRs,supplementing previous researches conducted at the populationcell level.The findings offered a reasonable risk analysis for the potential application of ZnO NRs in cancer therapy.This thesis holds significant reference value for the application of nanomaterials in biomedicine,particularly in the fields of cancer treatment and pathological analysis. |
