GLIPR1 modulates the response of cisplatin-resistant human lung cancer cells to cisplatin

PLoS One. 2017 Aug 3;12(8):e0182410. doi: 10.1371/journal.pone.0182410. eCollection 2017.

Abstract

Background and objective: Chemotherapy drugs, such as cisplatin (DDP), improve the survival of patients with lung cancer by inducing apoptosis in cancer cells, which quickly develop resistance to DDP through uncharacterized mechanisms. Glioma Pathogenesis-Related Protein 1 (GLIPR1) plays an important role in cell proliferation, migration and apoptosis. However, the expression and function of GLIPR1 in mediating DDP resistance in human lung adenocarcinoma A549/DDP and human large cell lung cancer H460/DDP cells has not yet been reported.

Methods: In this study, real-time PCR (RT-PCR) and western blot were used to examine the mRNA and protein expression of GLIPR1, respectively. Bright-field microscopy, the cell counting kit-8 (CCK-8) assay, flow cytometry analysis and JC-1 dye were used to measure the cellular morphology, proliferation, apoptosis and mitochondrial membrane potential, respectively.

Results: Compared to human lung adenocarcinoma A549 cells, the mRNA and protein expression of GLIPR1 were significantly increased in DDP-resistant A549/DDP cells (p < 0.05). Similarly, the mRNA level of GLIPR1 in DDP-resistant H460/DDP cells was also significantly higher than that in DDP-sensitive H460 cells (p < 0.05). Silencing of GLIPR1 in A549/DDP and H460/DDP cells led to increased apoptosis via a mitochondrial signaling pathway following incubation with various concentrations of DDP. Furthermore, GLIPR1 downregulation markedly reduced the protein expression of Bcl-2, and increased the cleaved Poly (ADP-Ribose) Polymerase (PARP) and cleaved caspase-3 in DDP-resistant A549/DDP cells.

Conclusion: In this study, we demonstrated for the first time that GLIPR1 could modulate the response of DDP-resistant A549/DDP and H460/DDP cells to cisplatin. Therefore, GLIPR1 deserves further investigation in the context of none-small lung cancer (NSCLC).

MeSH terms

  • Adenocarcinoma / drug therapy
  • Adenocarcinoma / metabolism
  • Adenocarcinoma / pathology*
  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects
  • Carcinoma, Large Cell / drug therapy
  • Carcinoma, Large Cell / metabolism
  • Carcinoma, Large Cell / pathology*
  • Cell Proliferation / drug effects
  • Cisplatin / pharmacology*
  • Drug Resistance, Neoplasm / genetics*
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Lung Neoplasms / drug therapy
  • Lung Neoplasms / metabolism
  • Lung Neoplasms / pathology*
  • Membrane Potential, Mitochondrial / drug effects
  • Membrane Proteins
  • Neoplasm Proteins / antagonists & inhibitors
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism*
  • Nerve Tissue Proteins / antagonists & inhibitors
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Poly(ADP-ribose) Polymerases / metabolism
  • RNA, Small Interfering / genetics
  • Tumor Cells, Cultured

Substances

  • Antineoplastic Agents
  • GLIPR1 protein, human
  • Membrane Proteins
  • Neoplasm Proteins
  • Nerve Tissue Proteins
  • RNA, Small Interfering
  • Poly(ADP-ribose) Polymerases
  • Cisplatin

Grants and funding

The grants and their roles in this study are listed below: 1. The 973 Program of the Ministry of Science and Technology of China (2012CB933304), playing a role in the design of the study; 2. The Natural Science Foundation of China (81570028 and 81400018), in data collection; 3. The Doctoral Program of Higher Education of China (20130071120063), in data analysis; 4. The National Key Scientific & Technology Support Program (2013BAI09B09), in data interpretation; 5. Youth Foundation of Zhongshan Hospital Fudan University (2015ZSQN26), in writing and editing the manuscript. 6. The Shanghai Committee of Science and Technology (12411950100), in the design of the study and data analysis.