Study On The Therapeutic Effect Of EpCAM-ING1-CART In Triple-negative Breast Cancer And Ovarian Cancer

Objective: To study the effects of EpCAM-ING1-CART on the treatment of triple-negative breast cancer and ovarian cancer.Method:Immunohistochemistry was used to detect the expression of EpCAM and EGFR in triple-negative breast cancer tissues and EpCAM in adjacent tissues.Flow cytometry was used to detect the expression of EpCAM in different human breast cancer cell lines and SKOV3 cells.The scFv of anti-EpCAM-CAR was designed based on the amino acid sequence of exon2 of EpCAM antigen,and called Ep CAM-ING1-CAR.The CAR-expressing T cells were generated by the Lentiviral packaging system and detected by flow cytometry.The Killing Assay was detected by flow cytometry and the release of IFN-γ was detected by ELISA.The EMT changes of SKOV3 induced by malignant ascites were detected by cell immunofluorescence.Results : Immunohistochemistry suggests that EpCAM and EGFR were both highly expressed in triple-negative breast cancer,and EpCAMexpression was particularly significant.Flow Cytometry revealed that EpCAM was highly expressed in MDA-MB-468 cells and SKOV3 cells,while partially expressed in MDA-MB-231 cells.IFN-γdetected by ELISA showed that when co-cultured with MDA-MB-468 and SKOV3,CAR-T cells has higher release of IFN-γthan Control-T.The killing assay detected by flow cytometry suggested the tumor killing effect of EpCAM-ING1-CART on both EpCAM-high-expressing MDA-MB-468 cells and EpCAM-low-expressing MDA-MB-231 cells.Cell immunofluorescence confirmed the EMT changes happened in SKOV3 cells induced by malignant ascites.Conclusion: EpCAM-ING1-CART works in the treatment of triple-negative breast cancer and ovarian cancer.