Exploring Trastuzumab Resistance Biomarkers for HER2-positive Breast Cancer Based on Targeted Metabolomics of TCA-Related Metabolites
OBJECTIVE To investigate the levels of tricarboxylic acid cycle(TCA)-related metabolites in human epidermal growth factor receptor 2(HER2)-positive breast cancer trastuzumab-resistant and trastuzumab-sensitive cells,as well as to screen po-tential biomarkers of trastuzumab resistance.METHODS Targeted metabolomics was adopted to detect TCA-related metabolite levels including citrate,cis-aconitate,isocitrate,alpha-ketoglutarate,succinic acid,fumaric acid,malic acid and oxaloacetic acid based on ul-tra-high-performance liquid chromatography-mass spectrometry(UPLC-MS/MS)in trastuzumab-resistant and trastuzumab-sensitive cells.The linearity,accuracy and precision of the method were validated.Then,orthogonal partial least-squares discrimination analysis(OPLS-DA)was used to compare the relative differential expression in trastuzumab-resistant and trastuzumab-sensitive cells.Differen-tially expressed metabolites were screened according to P＜0.05,fold change(FC)＞1.5 or FC＜0.67.Finally,a multivariable mod-el based on SVM and receiver operating characteristic(ROC)curve analysis was used to assess the classification accuracy of each me-tabolite.RESULTS For all TCA-related metabolites,the standard curves were linear,the correlation coefficients(r)were greater than 0.994,the established method accuracy was 94％-105％and the precision coefficient of variation(CV,％)was less than 15％.Trastuzumab-resistant and trastuzumab-sensitive cells were clearly separated from each other in the OPLS-DA model.Malic acid,succinic acid and fumaric acid were significantly upregulated in trastuzumab-resistant cells(P＜0.05)compared to trastuzumab-sensitive cells.The areas under the curves(AUCs)of malic acid,succinic acid and fumaric acid were greater than 0.9 and the SVM model yielded the highest AUC of 1.000.CONCLUSION TCA-related metabolites are differentially expressed in trastuzumab-resist-ant and trastuzumab-sensitive cells.Succinic acid,fumaric acid and malic acid can serve as biomarkers that may provide potential val-ue for the diagnosis of trastuzumab resistance in HER2-positive breast cancer.
HER2-positive breast cancertrastuzumab-resistanceTCA-related metabolitytargeted metabolomicbiomarker
万方数据
维普
