1. Academic Validation
  2. LAP3 contributes to IFN-γ-induced arginine depletion and malignant transformation of bovine mammary epithelial cells

LAP3 contributes to IFN-γ-induced arginine depletion and malignant transformation of bovine mammary epithelial cells

  • BMC Cancer. 2022 Aug 8;22(1):864. doi: 10.1186/s12885-022-09963-w.
Li Li  # 1 Fengyang Li  # 2 Xiuhong Hu  # 1 3 Zengshuai Wu 2 Wenbo Ren 1 Tingting Wang 1 Zhengchao Ji 1 Na Li 2 Jingmin Gu 2 Changjiang Sun 2 Xin Feng 2 Wenyu Han 2 Jing Huang 4 Liancheng Lei 5
Affiliations

Affiliations

  • 1 Department of First Hospital, Jilin University, Xinmin Street 1, Changchun, China.
  • 2 State Key Laboratory for Zoonotic Diseases, College of Veterinary Medicine, Jilin University, Xi'an Road 5333, Changchun, China.
  • 3 Shannan Hospital, Shannan, China.
  • 4 Department of First Hospital, Jilin University, Xinmin Street 1, Changchun, China. huangj@jlu.edu.cn.
  • 5 State Key Laboratory for Zoonotic Diseases, College of Veterinary Medicine, Jilin University, Xi'an Road 5333, Changchun, China. leiliancheng@163.com.
  • # Contributed equally.
Abstract

Background: IFN-γ has been traditionally recognized as an inflammatory cytokine that involves in inflammation and autoimmune diseases. Previously we have shown that sustained IFN-γ induced malignant transformation of bovine mammary epithelial cells (BMECs) via arginine depletion. However, the molecular mechanism underlying this is still unknown.

Methods: In this study, the Amino acids contents in BMECs were quantified by a targeted metabolomics method. The acquisition of differentially expressed genes was mined from RNA-seq dataset and analyzed bioinformatically. Quantitative reverse transcription polymerase chain reaction (qRT-PCR), enzyme-linked immunosorbent assay (ELISA), western blotting, and immunohistochemistry (IHC) assay were performed to detect gene mRNA and protein expression levels. CCK-8 and would healing assays were used to detect cell proliferation and migration abilities, respectively. Cell cycle phase alternations were analyzed by flow cytometry.

Results: The targeted metabolomics analysis specifically discovered IFN-γ induced arginine depletion through accelerating arginine catabolism and inhibiting arginine anabolism in BMECs. Transcriptome analysis identified leucine Aminopeptidase 3 (LAP3), which was regulated by p38 and ERK MAPKs, to downregulate arginine level through interfering with argininosuccinate synthetase (ASS1) as IFN-γ stimulated. Moreover, LAP3 also contributed to IFN-γ-induced malignant transformation of BMECs by upregulation of HDAC2 (histone deacetylase 2) expression and promotion of cell cycle proteins cyclin A1 and D1 expressions. Arginine supplementation did not affect LAP3 and HDAC2 expressions, but slowed down cell cycle process of malignant BMECs. In clinical samples of patients with breast Cancer, LAP3 was confirmed to be upregulated, while ASS1 was downregulated compared with healthy control.

Conclusions: These results demonstrated that LAP3 mediated IFN-γ-induced arginine depletion to malignant transformation of BMECs. Our findings provide a potential therapeutic target for breast Cancer both in humans and dairy cows.

Keywords

ASS1; Arginine depletion; HDAC2; IFN-γ; LAP3; Malignant transformation.

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