MAb NJ001 inhibits lung adenocarcinoma invasiveness by directly regulating TIMP-3 promoter activity via FOXP1 binding sites.

Abstract

BACKGROUND:

Previously, we developed a monoclonal antibody (mAb) NJ001 that binds to the antigen SP70 in human non-small cell lung cancer (NSCLC) cells and showed it could inhibit lung adenocarcinoma (AD) growth. Here, we investigated the effect and mechanisms of NJ001 in lung AD metastasis.

METHODS:

Human lung AD cells (SPC-A1 and A549) were treated with different concentrations of mAb NJ001, and the effects of NJ001 on cell migration and invasive activity were investigated using wound-healing and Matrigel assays, respectively. The molecular mechanism of this inhibition was explored by microarrays, qRT-PCR, western blot, luciferase assays and electrophoretic mobility shift assays (EMSA).

RESULTS:

MAb NJ001 markedly suppressed lung AD cell migration; and the invasiveness of SPC-A1 and A549 cells treated with mAb NJ001 was diminished by 65%. Tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) was highly expressed in SPC-A1 cells treated with mAb NJ001, whereas knockdown of TIMP-3 by shRNA significantly increased SPC-A1 and A549 invasiveness. MAb NJ001 affects lung AD by inhibiting TIMP-3 through direct transcriptional regulation of FOXP1 binding sites in the TIMP-3 promoter region, as shown in luciferase assays and EMSA.

CONCLUSIONS:

MAb NJ001 inhibits invasiveness and metastasis in lung AD through the FOXP1 binding sites in the TIMP-3 promoter region. It may have clinical applications in preventing and treating metastatic lung AD.

• Authors: Chunrong Gu, Ying Luo, Shichang Zhang, Jian Xu, Jiexin Zhang, Huanyu Ju, Jingping Liu, Lixia Zhang, Yan Zhang, Lei Wu, Erfu Xie, Ting Xu, Shiyang Pan
• Journal: Thoracic cancer (Thorac Cancer) Vol. 11 Issue 9 Pg. 2630-2638 (09 2020) ISSN: 1759-7714 [Electronic] Singapore
• PMID: 32744429 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2020 The Authors. Thoracic Cancer published by China Lung Oncology Group and John Wiley & Sons Australia, Ltd.

Chemical References

• Antibodies, Monoclonal
• FOXP1 protein, human
• Forkhead Transcription Factors
• McAb NJ001
• Repressor Proteins
• TIMP3 protein, human
• Tissue Inhibitor of Metalloproteinase-3

Topics

• Adenocarcinoma of Lung (genetics, pathology)
• Antibodies, Monoclonal (metabolism)
• Binding Sites
• Cell Proliferation
• Forkhead Transcription Factors (metabolism)
• Humans
• Lung Neoplasms (genetics, pathology)
• Repressor Proteins (metabolism)
• Tissue Inhibitor of Metalloproteinase-3
• Transfection