title: [Combination of TLR7 agonist T7-ethacrynic acid conjugate with ROR1 has a stronger anti-breast cancer effect].
abstract: Objective To investigate the synergistic anti-breast cancer effect of Toll-like receptor 7 agonist T7-ethacrynic acid conjugate (T7-EA) in combination with receptor-tyrosine-kinase-like orphan receptor 1 (ROR1). Methods ROR1 cytotoxic T lymphocyte (CTL) epitope was predicted using Syfpeithi online software. Mouse spleen lymphocytes and bone marrow dendritic cells (DCs) were separately stimulated with 4 μmol/L T7-EA and 4 μmol/L ROR1 alone or in combination. ELISA assay was used to measure the levels of interferon-γ (IFN-γ), interleukin 12 (IL-12) and tumor necrosis factor-α (TNF-α). Xenograft model was established via subcutaneous injection of mouse breast cancer 4T1 cells. The mice were weekly treated through intraperitoneal administration of 3 mg/kg T7-EA, 15 mg/kg ROR1 or the combination of T7-EA and ROR1. After four rounds of treatment, tumor tissues were weighed. Serum level of anti-4T1 tumor protein IgG was measured by ELISA. Specific CTL activity was detected by lactate dehydrogenase (LDH) assay. Results The peptide PYCDETSSV was chosen as an antigen epitope of breast cancer. The T7-EA highly activated in vitro lymphocytes in a dose-dependent manner, which wasn't affected by other relevant peptides. The combination of T7-EA and ROR1 stimulated the secretion of IFN-γ and IL-12 by lymphocytes and TNF-α by bone marrow DCs. The growth of tumor in vivo was significantly inhibited by T7-EA combined with ROR1 compared with T7-EA or ROR1 alone. The specific CTL activity triggered by T7-EA combined with ROR1 was much stronger than that triggered by T7-EA or ROR1 alone. The titer of anti-4T1 tumor protein IgG induced by T7-EA combined with ROR1 was higher than that induced by T7-EA or ROR1. Conclusion The combination of T7-EA and ROR1 has a better killing effect on breast cancer.
authors.affiliation: Department of Pathophysiology, Laboratory of Stem Cell and Tissue Engineering, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Na
authors.initials: N
authors.lastname: Zhang
authors.affiliation: School of Medicine, Shenzhen University, Shenzhen 518060, China.
authors.forename: Guangyi
authors.initials: G
authors.lastname: Jin
authors.affiliation: School of Medicine, Shenzhen University, Shenzhen 518060, China.
authors.forename: Zhenchao
authors.initials: Z
authors.lastname: Jin
authors.affiliation: School of Medicine, Shenzhen University, Shenzhen 518060, China.
authors.forename: Bing
authors.initials: B
authors.lastname: Liu
authors.affiliation: School of Medicine, Shenzhen University, Shenzhen 518060, China.
authors.forename: Boya
authors.initials: B
authors.lastname: Peng
authors.affiliation: School of Medicine, Shenzhen University, Shenzhen 518060, China.
authors.forename: Ningning
authors.initials: N
authors.lastname: Gao
authors.affiliation: School of Medicine, Shenzhen University, Shenzhen 518060, China.
authors.forename: Yunlong
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Department of Pathophysiology, Laboratory of Stem Cell and Tissue Engineering, Chongqing Medical University, Chongqing 400016, China . *Corresponding author, E-mail: tangli@cqmu.edu.cn.
authors.forename: Li
authors.initials: L
authors.lastname: Tang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019496:Cancer Vaccines / Q000276:immunology* / Q000494:pharmacology; D045744:Cell Line, Tumor; D003713:Dendritic Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D004976:Ethacrynic Acid / Q000276:immunology*; D005260:Female; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D018664:Interleukin-12 / Q000276:immunology / Q000378:metabolism; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008325:Mammary Neoplasms, Experimental / Q000276:immunology* / Q000378:metabolism / Q000517:prevention & control; D008807:Mice, Inbred BALB C; D057050:Receptor Tyrosine Kinase-like Orphan Receptors / Q000276:immunology* / Q000378:metabolism; D013997:Time Factors; D051199:Toll-Like Receptor 7 / Q000819:agonists / Q000276:immunology* / Q000378:metabolism
chemical_list: D019496:Cancer Vaccines; D007074:Immunoglobulin G; C495348:TLR7 protein, human; D051199:Toll-Like Receptor 7; D018664:Interleukin-12; D007371:Interferon-gamma; C078716:ROR1 protein, human; D057050:Receptor Tyrosine Kinase-like Orphan Receptors; D004976:Ethacrynic Acid