Rapid Identification And Pharmacodynamic Study Of Breast Cancer By Single Cell Sers Technique

The incidence of breast cancer is increasing year by year and has become the highest morbidity cancer in the world.Rapid cancer screening and evaluation of therapeutic effect are the key to improve cancer survival rate and reduce mortality.Therefore,there is an urgent need for a simple and rapid method for rapid diagnosis and treatment monitoring of cancer in order to improve the survival rate of patients.Surface enhanced Raman spectroscopy（SERS）is an emerging spectroscopic analysis technology,which has the fast,sensitive,"fingerprint"characteristics,and can detect information at the molecular level.In this study,single-cell SERS technique was used to quickly distinguish breast cancer cells from normal cells and breast cancer cells with different molecular types.Based on this,the anticancer drug epirubicin was used to treat breast cancer cells,and SERS technology was used to further explore the drug targets and pharmacokinetics,so as to provide important reference data for rapid cancer screening,chemotherapy effect evaluation and tumor heterogeneity research.First of all,gold nanoparticles are used as the reinforced substrate,by means of unlabeled and direct addition of Au NPs,the binding mode between gold particles and cells was observed by confocal micro-Raman spectrometer,and the enhancement of Raman signals of cells by gold particles with different particle sizes.It was found that after directly add Au NPs to adherent cells,some gold particles would enter into the cell through cellular internalization,and after just a few minutes,the Raman signal of the whole cell was greatly enhanced,and the rapid detection technique of single cell SERS was successfully established and optimized,which laid a foundation for follow-up research.Secondly,the established single cell SERS technique was used to detect different types of breast cancer cells.It was found that different types of cells had unique Raman phenotypes.Combined with PCA-LDA and OPLS-DA analysis,breast cancer cells,normal breast epithelial cells and different types of breast cancer cells could be identified quickly and accurately,with an accuracy of more than 80%.It provides a rapid,lable-free and non-destructive method for cancer screening and typing diagnosis.Finally,the breast cancer cells were treated with epirubicin,and the SERS profiles of adherent single cells treated with epirubicin at different times were collected,then the spectral signals were mined and analyzed by OPLS-DA.The research results show that SERS-OPLS-DA can sensitively and accurately distinguish between drug-treated and untreated cells before the cell morphology has not changed significantly after the drug is treated for a short time.With the extension of drug action time,most adherent cells exfoliated,died and eliminated,but there were still a few cells that could"tolerate"drug treatment,and drug-resistant cells showed a SERS phenotype similar to that of untreated cells,indicating the atypical characteristics of breast cancer cells.It is very interesting that the treatment of breast cancer cells with epirubicin for 12 hours causes the characteristic response of biological macromolecules,in which the Raman characteristic peaks of nucleic acids(724 cm^-1,735 cm^-1)were the most sensitive to epirubicin treatment,and the characteristic peaks change significantly,suggesting that the target of epirubicin is nucleic acid molecules,which is consistent with other literature reports.It is further suggested that single cell SERS technique is an effective technical method for screening drug targets.In summary,this study established and optimized the single cell SERS detection technique.Using this technique combined with multivariate statistical analysis,we successfully realized the rapid identification of breast cancer cells from normal cells,as well as the effective differentiation of breast cancer cells with different molecular types,quickly identified the intracellular targets of anticancer drugs from the point of view of spectroscopy,and deeply analyzed the dynamic spectral characteristics of drug-cell interaction.This provides a new research idea for rapid cancer screening,drug screening and target prediction.