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Development and characterization of TrMab?6, a novel anti?TROP2 monoclonal antibody for antigen detection in breast cancer.

ABSTRACT: Trophoblast cell?surface antigen 2 (TROP2) is a type I transmembrane glycoprotein that is overexpressed in a number of cancer types, including triple?negative breast cancer. The current study aimed to develop a highly sensitive and specific monoclonal antibody (mAb) targeting TROP2, which could be used to evaluate TROP2 expression using flow cytometry, western blot analysis and immunohistochemistry by employing the Cell?Based Immunization and Screening (CBIS) method. The established anti?TROP2 mAb, TrMab?6 (mouse IgG_2b, ?), detected TROP2 on PA?tagged TROP2?overexpressing Chinese hamster ovary?K1 (CHO/TROP2?PA) and breast cancer cell lines, including MCF7 and BT?474 using flow cytometry. Western blot analysis indicated a 40 kDa band in lysates prepared from CHO/TROP2?PA, MCF7 and BT?474 cells. Furthermore, TROP2 in 57/61 (93.4%) of the breast cancer specimens was strongly detected using immunohistochemical analysis with TrMab?6. In conclusion, the current study demonstrated that TrMab?6 may be a valuable tool for the detection of TROP2 in a wide variety of breast cancer types.

SUBMITTER: Sayama Y

PROVIDER: S-EPMC7723163 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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Development and characterization of TrMab‑6, a novel anti‑TROP2 monoclonal antibody for antigen detection in breast cancer.

Sayama Yusuke Y Kaneko Mika K MK Kato Yukinari Y

Molecular medicine reports 20201210 2

Trophoblast cell‑surface antigen 2 (TROP2) is a type I transmembrane glycoprotein that is overexpressed in a number of cancer types, including triple‑negative breast cancer. The current study aimed to develop a highly sensitive and specific monoclonal antibody (mAb) targeting TROP2, which could be used to evaluate TROP2 expression using flow cytometry, western blot analysis and immunohistochemistry by employing the Cell‑Based Immunization and Screening (CBIS) method. The established anti‑TROP2 m ...[more]

PMID: 33300065

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Project description:Autophagy has been emerging as a novel cytoprotective mechanism to increase tumor cell survival under conditions of metabolic stress and hypoxia as well as to escape chemotherapy-induced cell death. To elucidate whether autophagy might also protect cancer cells from the growth inhibitory effects of targeted therapies, we evaluated the autophagic status of preclinical breast cancer models exhibiting auto-acquired resistance to the anti-HER2 monoclonal antibody trastuzumab (Tzb). We first examined the basal autophagic levels in Tzb-naive SKBR3 cells and in two pools of Tzb-conditioned SKBR3 cells (TzbR), which optimally grow in the presence of Tzb doses as high as 200 microg/ml Tzb. Fluorescence microscopic analyses revealed that the number of punctate LC3 structures -a hallmark of autophagy- was drastically higher in Tzb-refractory cells than in Tzb-sensitive SKBR3 parental cells. Immunoblotting analyses confirmed that the lipidation product of the autophagic conversion of LC3 was accumulated to high levels in TzbR cells. High levels of the LC3 lipidated form in Tzb-refractory cells were accompanied by decreased p62/sequestosome-1 protein expression, a phenomenon characterizing the occurrence of increased autophagic flux. Moreover, increased autophagy was actively used to survive Tzb therapy as TzbR pools were exquisitely sensitive to chemical inhibitors of autophagosomal formation/function. Knockdown of LC3 expression via siRNA similarly resulted in reduced TzbR cell proliferation and supra-additively interacted with Tzb to re-sensitize TzbR cells. Sub-groups of Tzb-naive SKBR3 parental cells accumulated LC3 punctate structures and decreased p62 expression after treatment with high-dose Tzb, likely promoting their own resistance. This is the first report showing that HER2-overexpressing breast cancer cells chronically exposed to Tzb exhibit a bona fide up-regulation of the autophagic activity that efficiently works to protect breast cancer cells from the growth-inhibitory effects of Tzb. Therapeutic targeting autophagosome formation/function might represent a novel molecular avenue to reduce the emergence of Tzb resistance in HER2-dependent breast carcinomas.

Dataset Information

Development and characterization of TrMab?6, a novel anti?TROP2 monoclonal antibody for antigen detection in breast cancer.

Publications

Development and characterization of TrMab‑6, a novel anti‑TROP2 monoclonal antibody for antigen detection in breast cancer.

Similar Datasets

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