TROP-2 exhibits tumor suppressive functions in cervical cancer by dual inhibition of IGF-1R and ALK signaling

Gynecol Oncol. 2019 Jan;152(1):185-193. doi: 10.1016/j.ygyno.2018.10.039. Epub 2018 Nov 12.

Abstract

Objective: Inactivation of tumor suppressor genes promotes initiation and progression of cervical cancer. This study aims to investigate the tumor suppressive effects of TROP-2 in cervical cancer cells and to explain the underlying mechanisms.

Methods: The tumor suppressive functions of TROP-2 in cervical cancer cells were examined by in vitro and in vivo tumorigenic functional assays. Downstream factors of TROP-2 were screened using Human Phospho-Receptor Tyrosine Kinase Array. Small molecule inhibitors were applied to HeLa cells to test the TROP-2 effects on the oncogenicity of IGF-1R and ALK. Protein interactions between TROP-2 and the ligands of IGF-1R and ALK were detected via immunoprecipitation assay and protein-protein affinity prediction.

Results: In vitro and in vivo functional assays showed that overexpression of TROP-2 significantly inhibited the oncogenicity of cervical cancer cells; while knockdown of TROP-2 exhibited opposite effects. Human Phospho-Receptor Tyrosine Kinase Array showed that the activity of IGF-1R and ALK was stimulated by TROP-2 knockdown. Small molecule inhibitors AG1024 targeting IGF-1R and Crizotinib targeting ALK were treated to HeLa cells with and without TROP-2 overexpression, and results from cell viability and migration assays indicated that the oncogenicity of vector-transfected cells was repressed to a greater extent by the inhibition of either IGF-1R or ALK than that of the TROP-2-overexpressed cells. Immunoprecipitation assay and protein-protein affinity prediction suggested protein interactions between TROP-2 and the ligands of IGF-1R and ALK.

Conclusions: Collectively, our results support that TROP-2 exhibits tumor suppressor functions in cervical cancer through inhibiting the activity of IGF-1R and ALK.

Keywords: ALK; Cervical cancer; IGF-1R; TROP-2; Tumor suppressor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anaplastic Lymphoma Kinase / antagonists & inhibitors*
  • Anaplastic Lymphoma Kinase / physiology
  • Animals
  • Antigens, Neoplasm / physiology*
  • Cell Adhesion Molecules / antagonists & inhibitors
  • Cell Adhesion Molecules / physiology*
  • Cell Proliferation
  • Female
  • HeLa Cells
  • Humans
  • Insulin-Like Growth Factor I / metabolism
  • Mice
  • Midkine / metabolism
  • Receptor, IGF Type 1
  • Receptors, Somatomedin / antagonists & inhibitors*
  • Receptors, Somatomedin / physiology
  • Signal Transduction / physiology
  • Tumor Suppressor Proteins / physiology*
  • Uterine Cervical Neoplasms / drug therapy
  • Uterine Cervical Neoplasms / pathology
  • Uterine Cervical Neoplasms / prevention & control*

Substances

  • Antigens, Neoplasm
  • Cell Adhesion Molecules
  • IGF1R protein, human
  • MDK protein, human
  • Receptors, Somatomedin
  • TACSTD2 protein, human
  • Tumor Suppressor Proteins
  • Midkine
  • Insulin-Like Growth Factor I
  • ALK protein, human
  • Anaplastic Lymphoma Kinase
  • Receptor, IGF Type 1