拟缺香茶菜水提物显著改善肿瘤细胞荧光成像分辨率OA

Fluorescence imaging is a common technique for investigating the subcellular localization of tar-get proteins and protein-protein interactions within the biomedical domain.However,the fluorescence imaging resolution of many cells is often suboptimal due to their thickness and volume.Expansion microscopy imaging technology represents a sophisticated solution to this problem,but the current protocol is characterized by operational complexity,limited stability,and necessity for relatively high concentrations of immunofluores-cence antibodies.Our previous studies found that Reniformin A,a compound isolated from Isodon excisoi-des,has a magnifying effect on tumor cells.In this study,a low-cost and easily operationally convenient method was explored to improve the resolution of fluorescence imaging.Firstly,the effects of the aqueous extract of I.excisoides on three tumor cell lines A549,MDA-MB-231,and HeLa were investigated across different concentrations and treatment durations.The results indicated that the aqueous extract of I.exci-soides induced a significant volumetric enlargement in A549 and MDA-MB-231 cells,effectively enhan-cing the fluorescence imaging resolution.The optimal treatment time and concentration were identified as 24 h and 8 mg/mL,respectively.Conversely,the extract showed no such effect on HeLa cells,suggesting its cell-specific mechanism of action.Secondly,morphological analysis of key organelles and the cytoskeleton was investigated after treatment with the aqueous extract.The results revealed that the extract did not influence the fundamental cytoskeletal structures(α-tubulin,F-actin)or the morphology and distribution characteristics of key organelles(lysosomes,mitochondria)in A549 and MDA-MB-231 cells.Thirdly,subcellular localization was performed with three proteins of known subcellular localization information(METTL6,SLC38A7 and MYH9)in A549 cells.The results showed that treatment with the aqueous extract did not alter the original localization characteristics of the three proteins and did not induce any nonspecific co-localization.Finally,using A549 cells as a model,immunofluorescence assays were conducted to preliminarily investigate the po-tential mechanism by which the aqueous extract enhances fluorescence imaging resolution in tumor cells.It is hypothesized that the extract may enhance integrin function,stabilize focal adhesions,and optimize focal adhesion dynamics,thereby providing improved structural support for increased cell volume.These results indicate that the aqueous extract at a high concentration(8 mg/mL)exerts an enlarging effect on certain tumor cells without affecting the morphology and distribution of the original cellular structure,thereby effectively improving the convenience and resolution of fluorescence imaging.The aqueous extract of I.excisoides has broad application potential in biomedical research.